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Office  of  Technology  Assessment 


Congressional  Board  of  the  97th  Congress 

TED  STEVENS,  Alaska,  Chairman 
MORRIS  K.  UDALL,  Arizona,  Vice  Chairman 


Senate 

ORRIN  G.  HATCH 
Utah 

CHARLES  McC.  MATHIAS,  JR. 
Maryland 

EDWARD  M.  KENNEDY 
Massachusetts 

ERNEST  F.  HOLLINGS 
South  Carolina 

(One  To  Be  Appointed) 


House 

GEORGE  E.  BROWN,  JR 
California 

JOHN  D.  DINGELL 
Michigan 

LARRY  WINN,  JR. 
Kansas 

CLARENCE  E.  MILLER 
Ohio 

COOPER  EV  ANS 
Iowa 


JOHN  H.  GIBBONS 
(Nonvoting) 


CHARLES  N.  KIMBALL,  Chairman 
Midwest  Research  Institute 

JEROME  B.  WIESNER,  Vice  Chairman 
Massachusetts  Institute  of  Technology 

J.  FRED  BUCY 
Teyas  Instruments,  Inc. 

CLAIRE  T.  DEDRICK 
California  Public  Utilities 


Advisory  Council 

JAMES  C.  FLETCHER 
Burroughs  Corp. 

S.  DAVID  FREEMAN 
Tennessee  Valley  Authority 

GILBERT  GUDE 
Library  of  Congress 

CARLN.  HODGES 


RACHEL  McCULLOCH 
University  of  Wisconsin 

FREDERICK  t:.  ROBBINS 
Institute  of  Medicine 

ELMER  B.  S TAA  I S 
General  Accounting  Office 

LEWIS  niOMAS 


University  of  Arizona  Memorial  Sloan-Kettering  Cancer  ( rnir 


Director 

JOHN  H.  GIBBONS 


The  Technology  Assessment  Board  approves  the  release  of  this  report.  The  v iews  ex|)ressed  in  this  t 
not  necessarily  those  of  the  Board,  OTA  Advisory  Council,  or  of  individual  memhers  thereof. 


Impacts 

« « 

of  Applied  Genetics 

Micro-Organisms,  Plants,  and  Animals 


OTA  Reports  are  the  principal  documentation  of  formal  assessment  projects. 
These  projects  are  approved  in  advance  by  the  Technology'  Assessment  Board.  At 
the  conclusion  of  a project,  the  Board  has  the  opportunity  to  review  the  report  but 
its  release  does  not  necessarily  imply  endorsement  of  the  results  by  the  Board  or  its 
individual  members. 


CONGRESS  OF  THE  UNITED  STATES 

Office  of  Technology  Assessment 

Washington,  0.  C.  20510 


^•S’\ 


Library  of  Congress  Catalog  Card  Number  81-600046 

For  sale  by  the  Superintendent  of  Documents, 

U.S.  Government  Printing  Office,  Washington,  D.C.  20402 


Foreword 


This  report  examines  the  application  of  classical  and  molecular  genetic  technol- 
ogies to  micro-organisms,  plants,  and  animals.  Congressional  support  for  an  assess- 
ment in  the  field  of  genetics  dates  back  to  1976  when  30  Representatives  requested  a 
study  of  recombinant  DNA  technologx’.  Letters  of  support  for  this  broader  study  came 
from  the  then  Senate  Committee  on  Human  Resources  and  the  House  Committee  on 
Interstate  and  Foreign  Commerce,  Subcommittee  on  Health  and  the  Environment. 

Current  developments  are  especially  rapid  in  the  application  of  genetic  technol- 
ogies to  micro-organisms;  these  were  studied  in  three  industries:  pharmaceutical, 
chemical,  and  food.  Classical  genetics  continue  to  play  the  major  role  in  plant  and 
animal  breeding  but  new  genetic  techniques  are  of  ever-increasing  importance. 

This  report  identifies  and  discusses  a number  of  issues  and  options  for  the  Con- 
gress, such  as: 

• Federal  Gox  ernment  support  of  R&D, 

• methods  of  improving  the  germplasm  of  farm  animal  species, 

• risks  of  genetic  engineering, 

• patenting  li\  ing  organisms,  and 

• public  invoh  ement  in  decisionmaking. 

The  Office  of  Technolog\'  Assessment  was  assisted  by  an  advisory  panel  of  scien- 
tists, industrialists,  labor  representatives,  and  scholars  in  the  fields  of  law,  economics, 
and  those  concerned  with  the  relationships  between  science  and  society.  Others  con- 
tributed in  two  workshops  held  during  the  course  of  the  assessment.  The  first  was  to 
investigate  public  perception  of  the  issues  in  genetics;  the  second  examined  genetic 
applications  to  animals.  Sixty  reviewers  drawn  from  universities.  Government,  in- 
dustry, and  the  law  prox  ided  helpful  comments  on  draft  reports.  The  Office  expresses 
sincere  appreciation  to  all  those  individuals. 

An  abbrex  iated  copy  of  the  summary  of  this  report  (ch.  1)  is  available  free  of 
charge  from  the  Office  of  Technology'  Assessment,  U.S.  Congress,  Washington,  D.C., 
20510.  In  addition,  the  xx  orking  papers  on  the  use  of  genetic  technology  in  human  and 
in  veterinary  medicine  are  ax  ailable  as  a separate  volume  from  the  National  Technical 
Information  Serx  ice. 


JOHN  H.  GIBBONS 
Director 


Impacts  of  Applied  Genetics  Advisory  Panel 


J.  E.  Legates,  Chairman 

Dean,  School  of  Agriculture  and  Life  Sciences,  North  Carolina  State  University 


Ronald  E.  Cape 
Cetus  Corp. 

Nina  V.  Fedoroff 
Department  of  Embryology 
Carnegie  Institution  of  Washington 

June  Goodfield 
The  Rockefeller  University 

Harold  P.  Green 

Fried,  Frank,  Harris,  Shriver  and  Kampelman 

Halsted  R.  Holman 
Stanford  University  Medical  School 

M.  Sylvia  Krekel 

Health  and  Safety  Office  " 

Oil,  Chemical,  and  Atomic  Workers 
International  Union 

Elizabeth  Kutter 
The  Evergreen  State  College 


Oliver  E.  Nelson,  Jr. 

Laboratory  of  Genetics 
University  of  Wisconsin 

David  Pimentel 
Department  of  Entomology 
Cornell  University 

Robert  Weaver 

Department  of  Agricultural  Economics 
Pennsylvania  State  University 

James  A.  Wright 
Pioneer  Hi-Bred  International 
Plant  Breeding  Division 

Norton  D.  Zinder 
The  Rockefeller  University 


iv 


Applied  Genetics  Assessment  Staff 


Joyce  C.  Lashof,  Assistant  Director,  OTA 
Health  and  Life  Sciences  Division 

(Jretchen  Kolsrud,  Program  Manager 
Zsolt  Harsanvi,  Project  Director 


Project  Staff 

Marva  Breznay,  Administrative  Assistant 
Lawrence  Burton,  Analyst 
Susan  CMymer,  Research  Assistant 
Renee  (L  Ford,*  Technical  Editor 
Michael  (lough.  Senior  Analyst 
Robert  (Irossmann,*  Analyst 
Richard  Hutton,*  Editor 
Geoffrey  M.  Karny,  Legal  Analyst 


Major  Contractors 

Beniamin  (1.  Brackett,  Unix  ersity  of  Pennsyh  ania 
The  Genex  Corp. 

W illiam  P.  O'Neill,  Poly-Planning  Serx  ices 
Plant  Resources  Institute 

Anthony  J.  Sinskey,  Massachusetts  Institute  of  Technology 
Aladar  A.  Szalay,  Boyce-Thompson  Institute 
\ irginia  W'albot,  Washington  Unix  ersity 


OTA  Publishing  Staff 

John  C.  Holmes,  Publishing  Officer 

John  Bergling*  Kathie  S.  Boss  Debra  iM.  Catcher 
Patricia  A.  Dyson*  Mary  Harvey*  Joe  Henson 


'O  r X contrarl  pefsonnel. 


Contents 


Page 

Glossary viii 

Acronyms  and  Abbreviations xii 

Chapter  1.  Summary:  Issues  and  Options 3 

Chapter  2.  Introduction 29 

Part  I;  Biotechnology 

Chapter  3.  Genetic  Engineering  and  the  Fermentation  Technologies 49 

Chapter  4.  The  Pharmaceutical  Industry 59 

Chapter  5.  The  Chemical  Industry 85 

Chapter  6.  The  Food  Processing  Industry 107 

Chapter  7.  The  Use  of  Genetically  Engineered  Micro-Organisms  in  the 

Environment 117 


Part  II:  Agriculture 

Chapter  8.  The  Application  of  Genetics  to  Plants 137 

Chapter  9.  Advances  in  Reproductive  Biology  and  Their  Effects  on 

Animal  Improvement .* 167 

Part  III:  Institutions  and  Society 

Chapter  10.  The  Question  of  Risk 199 

Chapter  11.  Regulation  of  Genetic  Engineering 

Chapter  12.  Patenting  Living  Organisms 237 

Chapter  13.  Genetics  and  Society' 

Appendixes 

I-A.  A Case  Study  of  Acetaminophen  Production  269 

I-B.  A Timetable  for  the  Commercial  Production  of  Compounds  Using 

Genetically  Engineered  iMicro-Organisms  in  Biotechnology 275 

I-C.  Chemical  and  Biological  Processes 292 

I- D.  The  Impact  of  Genetics  on  Ethanol— A Case  Study 293 

II- A.  A Case  Study  of  W^heat 304 

II-B.  Genetics  and  the  Forest  Products  Industry  Case  Study 307 

II- C.  Animal  Fertilization  Technologies 309 

III- A.  History  of  the  Recombinant  DNA  Debate 315 

III-B.  Constitutional  Constraints  on  Regulation 320 

III-C.  Information  on  International  Guidelines  for  Recombinant  DNA 322 

I\’.  Planning  Workshop  Participants.  Other  Contractors  and  Contributors,  and 

Acknowledgments 329 


VII 


Glossary 


Aerobic.— Growing  only  in  the  presence  of  oxygen. 

Anaerobic.— Growing  only  in  the  absence  of 
oxygen. 

Alkaloids.— A group  of  nitrogen-containing  organic 
substances  found  in  plants;  many  are  pharmaco- 
logically active— e.g.,  nicotine,  caffeine,  and 
cocaine. 

Allele.— Alternate  forms  of  the  same  gene.  For  ex- 
ample, the  genes  responsible  for  eye  color  (blue, 
brown,  green,  etc.)  are  alleles. 

Amino  acids.— The  building  blocks  of  proteins. 
There  are  20  common  amino  acids;  they  are 
joined  together  in  a strictly  ordered  "string” 
which  determines  the  character  of  each  protein. 

Antibody.— A protein  component  of  the  immune 
system  in  mammals  found  in  the  blood. 

Antigen.— A large  molecule,  usually  a protein  or 
carbohydrate,  which  when  introduced  in  the 
body  stimulates  the  production  of  an  antibody 
that  will  react  specifically  with  the  antigen. 

Aromatic  chemical.— An  organic  compound  con- 
taining one  or  more  six-membered  rings. 

Aromatic  polynjer. — Large  molecules  consisting 
of  repeated  structural  units  of  aromatic  chem- 
icals. 

Artificial  insemination. — The  manual  placement 
of  sperm  into  the  uterus  or  oviduct. 

Bacteriophage  (or  phage).— A virus  that  multi- 
plies in  bacteria.  Bacteriophage  lambda  is  com- 
monly used  as  a vector  in  recombinant  DNA  ex- 
periments. 

Bioassay.— Determination  of  the  relative  strength 
of  a substance  (such  as  a drug)  by  comparing  its 
effect  on  a test  organism  with  that  of  a standard 
preparation. 

Biomass.— Plant  and  animal  material. 

Biome. — A community  of  living  organisms  in  a ma- 
jor ecological  region. 

Biosynthesis. — The  production  of  a chemical  com- 
pound by  a living  organism. 

Biotechnology. — The  collection  of  industrial  proc- 
esses that  involve  the  use  of  biological  systems. 
For  some  of  these  industries,  these  processes  in- 


volve the  use  of  genetically  engineered  micro- 
organisms. 

Blastocyst.— An  early  developmental  stage  of  the 
embryo;  the  fertilized  egg  undergoes  sex'eral  cell 
divisions  and  forms  a hollow  ball  of  cells  called 
the  blastocyst. 

Callus.— The  cluster  of  plant  cells  that  results  from 
tissue  culturing  a single  plant  cell. 

Carbohydrates.— The  family  of  organic  molecules 
consisting  of  simple  sugars  such  as  glucose  and 
sucrose,  and  sugar  chains  (polysaccharides)  such 
as  starch  and  cellulose. 

Catalyst.— A substance  that  enables  a chemical 
reaction  to  take  place  under  milder  than  normal 
conditions  (e.g.,  lower  temperatures).  Biological 
catalysts  are  enzymes;  nonbiological  catalysts  in- 
clude metallic  complexes. 

Cell  fusion.— The  fusing  together  of  two  or  more 
cells  to  become  a single  cell. 

Cell  lysis.— Disruption  of  the  cell  membrane  allow- 
ing the  breakdown  of  the  cell  and  exposure  of  its 
contents  to  the  environment. 

Cellulase.— An  enzyme  that  degrades  cellulose  to 
glucose. 

Cellulose.— A polysaccharide  composed  entii  eh-  of 
several  glucose  units  linked  end  to  end;  it  consti- 
tutes the  major  part  of  cell  walls  in  plants. 

Chimera.- An  individual  composed  of  a mixture  of 
genetically  different  cells. 

Chloroplast.- The  structure  in  plant  cx'lls  w ln're 
photosynthesis  occurs. 

Chromosomes. — The  thread-like  (X)mpon('nt.s  of  a 
cell  that  are  composed  of  DNA  and  protein,  I hex 
contain  most  of  the  cell’s  DNA. 

Clone.— A group  of  genetically  identical  cells  or- 
organisms  asexually  descend(Kl  fiom  a common 
ancestor.  All  cells  in  the  clone  ha\('  the  same  g»-- 
netic  material  and  ai'e  exact  cojiies  of  the  original 

Conjugation.— The  one-way  ti'ansfer  of  DNA  be- 
tween bacteria  in  cellular  contact. 

Crossing-over.— A genetic  (;\ent  that  can  occur- 
during  celluar  replication,  which  irnoixcs  the 
breakage  and  lounion  of  DNA  molecules 

Cultivar. — An  or'ganism  dexeloped  and  perstslcnl 
under  cultivation. 


via 


Cytogenetics.— A branch  of  biolog\’  that  deals  with 
the  study  of  heredity  and  \ariation  hy  the  metli- 
ods  of  i)oth  cytology  (the  study  of  cells)  and 
genetics. 

Cytoplasm.— The  protoplasm  of  a cell,  e.xternal  to 
the  cell's  nuclear  memhrane. 

Diploid.— .A  cell  with  double  the  basic  chromosome 
number. 

D\A  (deoxyribonucleic  acid).— The  genetic  ma- 
terial found  in  all  li\  ing  organisms.  K\  ery  inher- 
ited characteristic  has  its  origin  somewhere  in 
the  code  of  each  indi\  idual's  complement  of  I3\',A. 


Gene.— The  hereditary  unit;  a segment  of  DNA 
coding  for  a specific  protein. 

Gene  expression.— The  manifestation  of  the  ge- 
netic material  of  an  organism  as  specific  traits. 

Genetic  drift.— Changes  of  gene  frequency  in  small 
population  due  to  chance  preserx  ation  or  extinc- 
tion of  particular  genes. 

Genetic  code.— The  biochemical  basis  of  heredity 
consisting  of  codons  (base  triplets  along  the  DNA 
se(iuence)  that  determine  the  specific  amino  acid 
sequence  in  proteins  and  that  are  the  same  for  all 
forms  of  life  studied  so  far. 


D\.-\  vector.— A \ehicle  for  transferring  DN.A  from 
one  cell  to  another. 

Dominant  gene.— .A  characteristic  whose  expres- 
sion pre\  ails  o\  er  alternati\  e characteristics  for  a 
gi\en  trait. 

Blscherichiit  coli.—.\  bacterium  that  commonly  in- 
habits the  human  intestine.  It  is  a fa\orite  orga- 
nism for  many  microbiological  experiments. 

Endotoxins.— Complex  molecules  (lipopolysaccha- 
rides)  that  compose  an  integral  part  of  the  cell 
wall,  and  are  released  only  when  the  integrity  of 
the  cell  is  disturbed. 

Embryo  transfer.— Implantation  of  an  embryo 
into  the  o\  iduct  or  uterus. 

Enzyme.— .A  functional  protein  that  catalyzes  a 
chemical  reaction.  Enzymes  control  the  rale  of 
metabolic  processes  in  an  organism;  they  are  the 
acti\  e agents  in  the  fermentation  process. 

Estrogens.— Female  sex  hormones. 

Estrus  (“heat”).— The  period  in  which  the  female 
will  allow  the  male  to  mate  her. 

Eukaryote.— A higher,  compartmentalized  cell 
characterized  by  its  extensive  internal  structure 
and  the  presence  of  a nucleus  containing  the 
DNA.  .All  multicellular  organisms  are  eukaryotic. 
The  simpler  cells,  the  prokaryotes,  ha\e  much 
less  compartmentalization  and  internal  struc- 
ture; bacteria  are  prokaryotes. 

Exotoxins.— Proteins  produced  by  bacteria  that  are 
able  to  diffuse  out  of  the  cells;  generally  more  po- 
tent and  specific  in  their  action  than  endotoxins. 

Fermentation. — The  biochemical  process  of  con- 
\erting  a raw  material  such  as  glucose  into  a 
product  such  as  ethanol. 

Fibroblast.— A cell  that  gives  rise  to  connective 
tissues. 

Gamete.— A mature  reproductive  cell. 


Genetic  engineering.— A technologv'  used  at  the 
laboratory  level  to  alter  the  hereditary  apparatus 
of  a li\  ing  cell  so  that  the  cell  can  produce  more 
or  different  chemicals,  or  perfoi  m completely 
new  functions.  These  altered  cells  are  then  used 
in  industrial  production. 

Gene  mapping.— Determining  the  relative  loca- 
tions of  different  genes  on  a gi\  en  chromosome. 

Genome.— The  basic  chromosome  set  of  an 
organism— the  sum  total  of  its  genes. 

Genotype.— The  genetic  constitution  of  an  individ- 
ual or  group. 

Germplasm.— The  total  genetic  variability  available 
to  an  organism,  represented  by  the  pool  of  germ 
cells  or  seed. 

Germ  cell.— The  sex  cell  of  an  organism  (sperm  or 
egg,  pollen  or  ovum).  It  differs  from  other  cells  in 
that  it  contains  only  half  the  usual  number  of 
chromosomes.  Germ  cells  fuse  during  fertiliza- 
tion. 

Glycopeptides.— Chains  of  amino  acids  with  at- 
tached carbohydrates. 

Glycoprotein.— A conjugated  protein  in  which  the 
nonprotein  group  is  a carbohydrate. 

Haploid.— A cell  with  only  one  set  (half  of  the  usual 
number)  of  chromosomes. 

Heterozygous.— When  the  two  genes  controlling  a 
particular  trait  are  different,  the  organism  is 
heterozygous  for  that  trait. 

Homozygous.— When  the  two  genes  controlling  a 
particular  trait  are  identical  for  a pair  of  chro- 
mosomes, the  organism  is  said  to  be  homozygous 
for  that  trait. 

Hormones.— The  "messenger”  molecules  of  the 
body  that  help  coordinate  the  actions  of  various 
tissues;  they  produce  a specific  effect  on  the  ac- 
tivity of  cells  remote  from  their  point  of  origin. 


ix 


Hybrid.— A new  variety  of  plant  or  animal  that  re- 
sults from  cross-breeding  two  different  existing 
varieties. 

Hydrocarbon.— All  organic  compounds  that  are 
composed  only  of  carbon  and  hydrogen. 

Immunoproteins.— All  the  proteins  that  are  part 
of  the  immune  system  (including  antibodies^  in- 
terferon, and  cytokines). 

In  vitro.— Outside  the  living  organism  and  in  an 
artificial  environment. 

In  vivo.— Within  the  living  organism. 

Leukocytes.— The  white  cells  of  blood. 

Lipids.— Water  insoluble  biomolecules,  such  as  cel- 
lular fats  and  oils. 

Lipopolysaccharides.— Complex  substances  com- 
posed of  lipids  and  polysaccharides. 

Lymphoblastoid.— Referring  to  malignant  white 
blood  cells. 

Lymphokines.— The  biologically  active  soluble  fac- 
tor produced  by  white  blood  cells. 

Maleic  anhydride.— An  important  organic  chem- 
ical used  in  the  manufacture  of  synthetic  resins, 
in  fungicides,  in  the  dyeing  of  cotton  textiles,,  and 
to  prevent  the  oxidation  of  fats  and  oils  during 
storage  and  rancidity. 

Messenger  RNA.— Ribonucleic  acid  molecules  that 
serve  as  a guide  for  protein  synthesis. 

Metabolism.— The  sum  of  the  physical  and  chem- 
ical processes  involved  in  the  maintenance  of  life 
and  by  which  energy  is  made  available. 

Mitochondria.— Structures  in  higher  cells  that 
serve  as  the  “powerhouse”  for  the  cell,  producing 
chemical  energy. 

Monoclonal  antibodies.— Antibodies  derived 
from  a single  source  or  clone  of  cells  which 
recognize  only  one  kind  of  antigen. 

Mutants.— Organisms  whose  visible  properties  with 
respect  to  some  trait  differ  from  the  norm  of  the 
population  due  to  mutations  in  its  DNA. 

Mutation. — Any  change  that  alters  the  sequence  of 
bases  along  tbe  DNA,  changing  the  genetic  ma- 
terial. 

Myeloma. — A malignant  disease  in  which  tumor 
cells  of  the  antibody  producing  system  synthesize 
excessive  amounts  of  specific  proteins. 

n-alkanes. — Straight  chain  hydrocarbons — the 

main  constituents  of  petroleum. 


Nif  genes.— The  genes  for  nitrogen  fixation  present 
in  certain  bacteria. 

Nucleic  acid.— A polymer  composed  of  DNA  or 
RNA  subunits. 

Nucleotides.— The  fundamental  units  of  nucleic 
acids.  They  consist  of  one  of  the  four  bases— 
adenine,  guanine,  cytosine,  and  thymine  (uracil 
in  the  case  of  RNA)— and  its  attached  sugar-phos- 
phate group. 

Organic  compounds.— Chemical  compounds 

based  on  carbon  chains  or  rings,  which  contain 
hydrogen,  and  also  may  contain  oxygen,  nitro- 
gen, and  various  other  elements. 

Parthenogenesis.— Reproduction  in  animals  with- 
out male  fertilization  of  the  egg. 

Pathogen.— A specific  causative  agent  of  disease. 

Peptide.— Short  chain  of  amino  acids. 

pH.-A  measure  of  the  acidity  or  basicity  of  a solu- 
tion; on  a scale  of  0 (acidic)  to  14  (basic):  for  exam- 
ple, lemon  juice  has  a pH  of  2.2  (acidic),  water  has 
a pH  of  7.0  (neutral),  and  a solution  of  baking 
soda  has  a pH  of  8.5  (basic). 

Phage.— (See  bacteriophage.) 

Phenotype.— Tbe  visible  properties  of  an  organism 
that  are  produced  by  the  interaction  of  the  geno- 
type and  the  environment. 

Plasmid.— Hereditary  material  that  is  not  part  of  a 
chromosome.  Plasmids  are  circular-  and  sc'lf-repli- 
cating.  Because  they  ai-e  gener  ally  srirall  ;md  rela- 
tively simple,  they  ar-e  used  in  r-ecornbinant  DN.A 
experiments  as  acceptor's  of  foreign  DN.A. 

Plastid.— Any  specialized  or-gan  of  the  plant  cell 
other  than  the  nucleus,  such  as  the  chloroplast 

Ploidy.— Describes  the  number  of  srUs  of  chromo- 
somes present  in  the  or-ganism.  I'or  example, 
humans  are  diploid,  having  two  hoiirologous  sets 
of  23  chromosomes  (one  set  fr-ont  each  parent) 
for  a total  of  48  chr'omosomes;  manv  plants  .ire 
haploid,  having  only  one  copy  of  each  chro- 
mosome. 

Polymer.— A long-chain  nrolecule  foiined  li'om 
smaller  repeating  structur-al  units. 

Polysaccharide.— A long-chain  carbohydrate  con- 
taining at  least  three  molecules  of  sim|)le  sug.irs 
linked  together;  examples  would  include  (ellu 
lose  and  star'ch. 

Progestogens.— Hormones  invoked  with  ovul.i 
tion. 


X 


Prosla^jlandin.— Refers  to  a group  of  naturally  oc- 
curring, chemically  related  long-chain  fatty  acids 
that  have  certain  physiological  effects  (stimulate 
contraction  of  uterine  and  other  smooth  muscles, 
lower  hlood  pressure,  affect  action  of  certain 
hormones). 

Protein.— .A  linear  polymer  of  amino  acids;  proteins 
are  the  products  of  gene  e.xpression  and  are  the 
functional  and  structural  components  of  cells. 

Protoplast.— ,A  cell  without  a wall. 

Protoplast  fusion.— A means  of  achieving  genetic 
transformation  by  joining  two  protoplasts  or  join- 
ing a protoplast  with  any  of  the  components  of 
another  cell. 

Kecessive  gene.— .Any  gene  whose  e.xpression  is 
dependent  on  the  absence  of  a dominant  gene. 

Recombinant  D\A.— The  hybrid  DN.A  produced 
by  joining  pieces  of  DN.A  from  different  sources. 

Restriction  enzyme.— An  enzyme  within  a bac- 
terium that  recognizes  and  degrades  DN.A  from 
foreign  organisms,  thereby  preserving  the  genet- 
ic integrity  of  the  bacterium.  In  recombinant 
DNA  e.xperiments,  restriction  enzymes  are  used 
as  tiny  biological  scissors  to  cut  up  foreign  DN.A 
before  it  is  recombined  with  a vector. 

Reverse  transcriptase.— .An  enzyme  that  can  syn- 
thesize a single  strand  of  DN.A  from  a messenger 


RNA,  the  re\  erse  of  the  normal  direction  of  proc- 
essing genetic  information  within  the  cell. 

RiN'.A  (ribonucleic  acid).— In  its  three  forms— mes- 
senger RN.A,  transfer  RN.A,  and  ribosomal  RNA— 
it  assists  in  translating  the  genetic  message  of 
DN.A  into  the  finished  protein. 

Somatic  cell.— One  of  the  cells  composing  parts  of 
the  body  (e.g.,  tissues,  organs)  other  than  a germ 
cell. 

'I’issue  culture.— .An  in  vitro  method  of  propagat- 
ing healthy  cells  from  tissues,  such  as  fibroblasts 
from  skin. 

Transduction.— The  pi'ocess  by  which  foreign 
DNA  becomes  incorporated  into  the  genetic  com- 
plement of  the  host  cell. 

Transformation.— The  transfer  of  genetic  infor- 
mation by  DNA  separated  from  the  cell. 

V'ector.— ,A  transmission  agent;  a DNA  vector  is  a 
self-replicating  DNA  molecule  that  transfers  a 
piece  of  DNA  from  one  host  to  another. 

V'irus.— An  infectious  agent  that  requires  a host  cell 
in  order  for  it  to  replicate.  It  is  composed  of 
either  RNA  or  DNA  wrapped  in  a protein  coat. 

Zygote. — A cell  formed  by  the  union  of  two  mature 
reproductive  cells. 


Acronyms  and  Abbreviations 


\.\ 

— amino  acids 

IBCs 

— Institutional  Biosafety  Committees 

\(  :s 

— American  Cancer  Society 

ICI 

— Imperial  Chemical  Industries 

Acni 

— adrenocorticotropic  hormone 

IND 

— Investigational  New  Drug  Application 

,\i 

— ai-tificial  insemination 

(FDA) 

AII’L 

— Animal  Improvement  Programs 

kg 

— kilogram 

Eahoratory 

1 

— liter 

Al'AP 

— acetaminophen 

lb 

— pound 

ASM 

— American  Society  for  Microbiology 

mg 

— milligram 

1)1)1 

— barrel(s) 

gg 

— microgram 

l)l)l/cl 

— barrels  per  day 

/tm 

— micrometer  (formerly  micron) 

liODf) 

-5-day  biochemical  oxygen  demand 

MUA 

— Memorandum  of  Understanding  and 

BKM 

— Biological  Response  Modifier  Program 

Agreement 

1)U 

— bushel 

NCI 

— National  Cancer  Institute 

CaMV' 

— cauliflower  mosaic  virus 

NDA 

— new  drug  application  (FDA) 

CCPA 

— The  Court  of  Customs  and  Patent 

NDAB 

— National  Diabetics  Ad\'isory  Board 

Appeals 

NDCHIP 

— National  Cooperative  Dairy  Herd 

c:dc: 

— Center  for  Disease  Control 

Program 

CERB 

— Cambridge  Experimentation  Review 
Board 

NIAID 

— National  Institute  of  Allergy  and 
Infectious  Diseases 

13HHS 

— Department  of  Health  and  Human 
Services  (formerly  Health,  Education, 

NIAMDD 

— National  Institute  of  Arthritis, 
Metabolism,  and  Digestix  e Diseases 

and  Welfare) 

NIH 

— National  Institutes  of  Health 

DHI 

— Dairy  Herd  Improvement 

NIOSH 

— National  Institute  of  Occupational 

DNA 

— deoxyribonucleic  acid 

Safety  and  Health 

DOC 

— Department  of  Commerce 

NSF 

— National  Science  Foundation 

UOD 

— Department  of  Defense 

OECD 

— The  Organization  for  Economic 

DOE 

— Department  of  Energy 

Cooperation  and  Dexelopment 

DRAG 

— Dangerous  Pathogens  Advisory  Group 

ORDA 

— Office  of  Recombinant  DNA  Actix  ities 

EOR 

— Enhanced  oil  recovery 

PD 

— predicted  difference 

EPA 

— Environmental  Protection  Agency 

pH 

— unit  of  measure  for  acidity/hasicity 

FDA 

— Food  and  Drug  Administration 

ppm 

— parts  per  million 

FMDV' 

— foot-and-mouth  disease  virus 

R&.D 

— research  and  dexelopment 

— square  foot 

RAC 

— Recombinant  DNA  Advisory  Commitit 

ft 

— foot 

rDNA 

— recombinant  DNA 

FTC 

— Federal  Trade  Commission 

SCP 

— single-cell  protein 

g 

— gram 

T-DNA 

— a smaller  segment  of  the  Fi  plasmid 

gal 

— gallon 

Ti 

— tumor  inducing 

GH 

— growth  hormone 

TSCA 

— Toxic  Substances  Control  Act 

ha 

— hectares 

UCSF 

— University  of  California  at  San 

HEW 

— Department  of  Health,  Education,  and 

Francisco 

Welfare 

U.S.C. 

— United  States  Code 

hGH 

— human  growth  hormone 

USDA 

— United  States  De|)artment  of 

HYV' 

— high-yielding  varieties 

Agriculture 

XII 


chapter  1 

Summary:  Issues  and  Options 


chapter  1 


Page 


Hiotechnology 4 

The  Pharmaceutical  industry 4 

Findings 4 

The  Chemical  Industry 7 

Findings 7 

Food  processing  industry 8 

Findings 8 

The  Use  of  Genetically  Engineered  Micro- 

Organisms  in  the  Environment 8 

Findings 8 

Mineral  Leaching  and  Recovery 9 

Enhanced  Oil  Recovery 9 

Pollution  Control 9 

Constraints  in  Using  Genetic  Engineering 

Technologies  in  Open  Environments 10 

Issue  and  Options— Biotechnology 10 

Agriculture 11 

The  Applications  of  Genetics  to  Plants 11 

Findings 11 

New  Genetic  Technologies  for  Plant  Breeding  12 
Constraints  on  Using  Molecular  Genetics 

for  Plant  Improvements 13 

Genetic  Variability,  Crop  Vulnerability,  and 

the  Storage  of  Germplasm 13 

Issues  and  Options— Plants 14 

Advances  in  Reproductive  Biology  and  Their 
Effects  on  Animal  Improvement 15 


Page 

Findings 15 

Issue  and  Options— Animals 17 

Institutions  and  Society 18 

Regulation  of  Genetic  Engineering 18 

Findings 18 

Issue  and  Options— Regulation 20 

Patenting  Living  Organisms 22 

Findings 22 

Issue  and  Options— Patenting  Living  Organisms  23 

Genetics  and  Society 24 

Issues  and  Options— Genetics  and  Society 24 


Table 

Table  No.  Page 

1.  Containment  Recommended  by  the  National 

Institutes  of  Health 19 

Figures 

Figure  No.  Page 

1.  Recombinant  DNA;  The  Technique  of 

flecombining  Genes  From  One  Species  With 
Those  From  Another 5 

2.  The  Product  Development  Process 8 

3.  The  Way  the  Reproductive  Technologies 

Interrelate 16 


chapter  1 

Summary:  Issues  and  Options 


The  genetic  alteration  of  plants,  animals,  and 
micro-organisms  has  been  an  important  part  of 
agriculture  for  centuries.  It  has  also  been  an  in- 
tegral part  of  the  alcoholic  beverage  industry 
since  the  invention  of  beer  and  wine:  and  for 
the  past  century,  a mainstay  of  segments  of  the 
pharmaceutical  and  chemical  industries. 

However,  only  in  the  last  20  years  have  pow- 
erful new  genetic  technologies  been  developed 
that  greatly  increase  the  ability  to  manipulate 
the  inherited  characteristics  of  plants,  animals, 
and  micro-organisms.  One  consequence  is  the 
increasing  reliance  the  pharmaceutical  and 
chemical  industries  are  placing  on  hiotechnol- 
ogv . Micro-organisms  are  being  used  to  manu- 
facture substances  that  have  previously  been 
e.xtracted  from  natural  sources.  .Animal  and 
plant  breeders  are  using  the  new  techniques  to 
help  clarify  basic  questions  about  biological 
functions,  and  to  improve  the  speed  and  effi- 
ciency of  the  technologies  they  already  use. 
Other  industries— from  food  processing  and  pol- 
lution control  to  mining  and  oil  recovery— are 
considering  the  use  of  genetic  engineering  to  in- 
crease productiv  ity  and  cut  costs. 

Genetic  technologies  will  have  a broad  impact 
on  the  future.  They  may  contribute  to  filling 
some  of  the  most  fundamental  needs  of  man- 
kind—from  health  care  to  supplies  of  food  and 
energv'.  At  the  same  time,  they  arouse  concerns 
about  their  potential  effects  on  the  environment 
and  the  risks  to  health  involved  in  basic  and 
applied  scientific  research  and  development 
(R&D).  Because  genetic  technologies  are  already 
being  applied,  it  is  appropriate  to  begin  con- 
sidering their  potential  consequences. 

Congressional  concern  w ith  applied  genetics 
dates  back  to  1976,  when  30  Representatives  re- 
quested an  assessment  of  recombinant  DNA 
(rDNA)  technology.  Support  for  the  broader 
study  reported  liere  came  in  letters  to  the  Office 
of  Technology  Assessment  from  the  then  Senate 
Committee  on  Human  Resources  and  the  House 
Committee  on  Interstate  and  Foreign  Com- 
merce, Subcommittee  on  Health  and  the  Envi- 


ronment. In  addition,  specific  subtopics  are  of 
interest  to  other  committees,  notably  those  hav- 
ing jurisdiction  over  science  and  technology  and 
those  concerned  with  patents. 

This  report  describes  the  potentials  and  prob- 
lems of  applying  the  new  genetic  technologies  to 
a range  of  major  industries.  It  emphasizes  the 
present  state  of  the  art  because  that  is  what 
defines  the  basis  for  the  future  applications.  It 
then  makes  some  estimates  of  economic,  envi- 
ronmental, and  institutional  impacts— where, 
when,  and  how  some  technologies  might  be  ap- 
plied and  what  some  of  the  results  might  be. 
The  report  closes  with  the  possible  roles  that 
Government,  industry,  and  the  public  might 
play  in  determining  the  future  of  applied 
genetics. 

The  term  applied  genetics,  as  used  in  this 
report,  refers  to  two  groups  of  technologies: 

• Classical  genet/cs— natural  mating  methods 
for  the  selective  breeding  of  organisms 
for  desired  characteristics— e.g.,  breeding 
cows  for  increased  milk  production.  The 
pool  of  genes  available  for  selection  is  com- 
prised of  those  that  cause  natural  differ- 
ences among  individuals  in  a population 
and  those  obtained  by  mutation. 

• Molecular  genetics  includes  the  technologies 
of  genetic  engineering  that  involve  the 
directed  manipulation  of  the  genetic  mate- 
rial itself.  These  technologies — such  as 
rDNA  and  the  chemical  synthesis  of  genes 
—can  increase  the  size  of  the  gene  pool  for 
any  one  organism  by  making  available  ge- 
netic traits  from  many  different  popula- 
tions. Molecular  genetics  also  includes 
technologies  in  which  manipulation  occurs 
at  a level  higher  than  that  of  the  gene— at 
the  cellular  level,  e.g.,  cell  fusion  and  in 
vitro  fertilization. 

Significant  applications  of  molecular  genetics 
to  micro-organisms,  such  as  the  efforts  to  man- 
ufacture human  insulin,  are  already  underway 
in  several  industries.  Most  of  these  applications 


3 


4 • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


ciopend  on  fermentation— a technology  in  which 
substances  produced  by  micro-organisms  can 
l)e  obtained  in  large  quantities.  Applications  to 


plants  and  animals,  which  are  biologically  more 
complex  and  more  difficult  to  manipulate  suc- 
cessfully, will  take  longer  to  develop. 


Biotechnology  

Biotechnology— the  use  of  living  organisms  or 
their  components  in  industrial  processes— is 
possible  because  micro-organisms  naturally  pro- 
duce countless  substances  during  their  lives. 
Some  of  these  substances  have  proved  commer- 
cially valuable.  A number  of  different  industries 
ha\  e learned  to  use  micro-organisms  as  natural 
factories,  cultivating  populations  of  the  best 
producers  under  conditions  designed  to  en- 
hance their  abilities. 

Applied  genetics  can  play  a major  role  in  im- 
proving the  speed,  efficiency,  and  productivity 
of  these  biological  systems.  It  permits  the  ma- 
nipulation, or  engineering,  of  the  micro-orga- 
nisms’ genetic  material  to  produce  the  desired 
characteristics.  Genetic  engineering  is  not  in 
itself  an  industry,  but  a technique  used  at  the 
laboratory  level  that  allows  the  researcher  to 
modify  the  hereditary  apparatus  of  the  cell.  The 
population  of  altered  identical  cells  that  grows 
from  the  first  changed  micro-organism  is,  in 
turn,  used  for  various  industrial  processes.  (See 
figure  1.) 

The  first  major  commercial  effects  of  the  ap- 
plication of  genetic  engineering  will  be  in  the 
pharmaceutical,  chemical,  and  food  processing 
industries.  Potential  commercial  applications  of 
value  to  the  mining,  oil  recovery,  and  pollution 
control  industries— which  may  desire  to  use  ma- 
nipulated micro-organisms  in  the  open  environ- 
ment-are still  somewhat  speculative. 

The  pharmaceutical  industry 
FINDINGS 

The  pharmaceutical  industry  has  been  the 
first  to  take  advantage  of  the  potentials  of  ap- 
plied molecular  genetics.  Ultimately,  it  will 
probably  benefit  more  than  any  other,  with  the 
largest  percentage  of  its  products  depending  on 
advances  in  genetic  technologies.  Already, 


micro-organisms  have  been  engineered  to  pro- 
duce human  insulin,  interferon,  growth  hor- 
mone, urokinase  (for  the  treatment  of  blood 
clots),  thymosin-a  1 (for  controlling  the  immune 
response),  and  somatostatin  (a  brain  hormone). 
(See  figure  2.) 

The  products  most  likely  to  be  affected  by 
genetic  engineering  in  the  next  10  to  20  years 
are  nonprotein  compounds  like  most  antibiotics, 
and  protein  compounds  such  as  enzymes  and 
antibodies,  and  many  hormones  and  \ accines. 
Improvements  can  be  made  both  in  the  prod- 
ucts and  in  the  processes  by  which  they  are  pro- 
duced. Process  costs  may  be  lowered  and  even 
entirely  new  products  developed. 

The  most  advanced  applications  today  are  in 
the  field  of  hormones.  While  certain  hormones 
have  already  proved  useful,  the  testing  of 
others  has  been  hindered  by  their  scarcit\'  and 
high  cost.  Of  48  human  hormones  that  ha\c 
been  identified  so  far  as  possible  candidates  for 
production  by  genetically  engineered  mici'o- 
organisms,  only  10  are  used  in  current  medical 
practice.  The  other  38  are  not,  j)artly  hc'cause 
they  have  been  available  in  such  limited  (|uan- 
tities  that  tests  of  their  therapeutic  \alue  ha\(> 
not  been  possible. 

Genetic  technologies  also  open  up  lunv  ap- 
proaches for  vaccine  development  for  such  in- 
tractable parasitic  and  viral  diseases  as  aiiK'hic 
dysentery,  trachoma,  hepatitis,  and  malaria.  ,\t 
present,  the  vaccine  most  likely  to  h(?  produced 
is  for  foot-and-mouth  disease  in  animals.  How  - 
ever, should  any  one  of  the  \ accin(!s  foi-  liimian 
diseases  become  available,  the  social,  economic, 
and  political  consequences  of  a d(U'reas(*  in  mor- 
bidity and  mortality  would  he  significant.  .Main 
of  these  diseases  are  particularly  i)re\alcnt  in 
less  industrialized  countries;  the?  dc\ ('li)|)mcnts 
of  vaccines  for  them  may  profoundly  affect  the 
lives  of  tens  of  millions  of  people. 


Ch.  1 Summary:  Issues  and  Options  • 5 


Figure  1.— Recombinant  DNA:  The  Technique  of  Recombining  Genes 
From  One  Species  With  Those  From  Another 


Electron  micrograph  of  the  DNA,  which  is  the  plasmid  SP01 
from  Bacillus  subtilis.  This  plasmid  which  has  been 
sliced  open  is  used  for  recombinant  DNA  research 
in  this  bacterial  host 


amount  of  DNA  protein 
SOURCE:  Office  of  Technology  Assessment. 

For  some  pharmaceutical  products,  biotech- 
nology will  compete  with  chemical  synthesis 
and  extraction  from  human  and  animal  organs. 
Assessing  the  relative  worth  of  each  method 
must  be  done  on  a case-by-case  basis.  But  for 
other  products,  genetic  engineering  offers  the 
only  method  known  that  can  ensure  a plentiful 
supply;  in  some  instances,  it  has  no  competition. 

By  making  a pharmaceutical  available,  genet- 
ic engineering  may  have  two  types  of  effects: 

• Drugs  that  already  have  medical  promise 


Photo  credits:  Professor  F.  A.  Eiserling,  UCLA  Molecular  Biology  Institute 


Electron  micrograph  of  Bacillus  subtilis  in  the  process  of 
cell  division.  The  twisted  mass  in  the  center  of  each 
daughter  cell  is  the  genetic  material,  DNA 


Restriction  enzymes  recognize  certain  sites  along  the  DNA 
and  can  chemically  cut  the  DNA  at  those  sites.  This  makes 
it  possible  to  remove  selected  genes  from  donor  DNA  mole- 
cules and  insert  them  into  plasmid  DNA  molecules  to  form 
the  recombinant  DNA.  This  recombinant  DNA  can  then  be 
cloned  in  its  bacterial  host  and  large  amounts  of  a desired 
protein  can  be  produced. 


will  be  available  in  ample  amounts  for  clin- 
ical testing.  Interferon,  for  example,  can  be 
tested  for  its  efficacy  in  cancer  and  viral 
therapy,  and  human  growth  hormone  can 
be  evaluated  for  its  ability  to  heal  wounds. 

• Other  pharmacologically  active  substances 
for  which  no  apparent  use  now  exists  will 
be  available  in  sufficient  quantities  and  at 
low  enough  cost  to  enable  researchers  to 
explore  new  uses.  As  a result,  the  potential 
for  totally  new  therapies  exists.  Regulatory 
proteins,  for  example,  which  are  an  entire 


6 • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Figure  2.— The  Product  Development  Process 


Micro-organisms  such  as  E.  coli 


20.  Submit  INO 


The  development  process  begins  by  obtaining  DNA  either  through  organic  synthesis  (1)  or  derived  from  biological  sources  such  as  tissues 
(2).  The  DNA  obtained  from  one  or  both  sources  is  tailored  to  form  the  basic  “gene"  (3)  which  contains  the  genetic  Information  to  "code"  tor  a 
desired  product,  such  as  human  interferon  or  human  insulin.  Control  signals  (4)  containing  instructions  are  added  to  this  gene  (5).  Circular  DNA 
molecules  called  plasmids  (6)  are  isolated  from  micro-organisms  such  as  E.  coli-,  cut  open  (7)  and  spliced  back  (8)  together  with  genes  and  con- 
trol signals  to  form  “recombinant  DNA”  molecules.  These  molecules  are  then  introduced  into  a host  ceii  (9). 

Each  piasmid  is  copied  many  times  in  a cell  (10).  Each  cell  then  translates  the  information  contained  in  these  plasmids  Into  the  desired  prod- 
uct, a process  calied  “expression”  (11).  Cells  divide  (12)  and  pass  on  to  their  offspring  the  same  genetic  information  contained  in  the  parent 
cell. 

Fermentation  of  large  populations  of  geneticaliy  engineered  micro-organisms  is  first  done  in  shaker  fiasks  (13),  and  then  in  small  fermenters 
(14)  to  determine  growth  conditions,  and  eventually  in  larger  fermentation  tanks  (15).  Cellular  extract  obtained  from  the  fermentation  process  la 
then  separated,  purified  (16),  and  packaged  (17)  either  for  industrial  use  (18)  or  health  care  applications. 

Health  care  products  are  first  tested  in  animal  studies  (19)  to  demonstrate  a product’s  pharmacological  activity  and  safety.  In  the  United 
States,  an  investigational  new  drug  application  (20)  is  submitted  to  begin  human  clinical  trials  to  establish  safety  and  efficacy  Following 
clinical  testing  (21),  a new  drug  application  (NDA)  (22)  is  filed  with  the  Food  and  Drug  Administration  (FDA).  Vi/hen  the  NDA  has  been  reviewed 
and  approved  by  the  FDA  the  product  may  be  marketed  in  the  United  States  (23). 


SOURCE:  Genentech,  Inc, 


Ch.  1 — Summary:  Issues  and  Options  • 7 


class  of  molecules  that  control  gene  acti\  i- 
ty,  are  present  in  the  body  in  only  minute 
quantities.  Now,  for  the  first  time,  they  can 
be  recognized,  isolated,  characterized,  and 
produced  in  cjuantity. 

The  mere  a\  ailahilit\’  of  a pharmacologically 
acti\  e substance  does  not  ensure  its  adoption  in 
medical  practice.  E\en  if  it  is  shown  to  have 
therapeutic  usefulness,  it  may  not  succeed  in 
the  market{)lace. 

The  difficulty  in  predicting  the  economic  im- 
pact is  e.xemplified  by  interferon.  If  it  is  found  to 
be  broadly  effecti\e  against  both  \ iral  diseases 
and  cancers,  sales  would  he  in  the  tens  of  bil- 
lions of  dollars  annually.  If  its  clinical  effec- 
tiveness is  found  to  be  only  against  one  or  two 
\ iruses,  sales  would  be  significantly  lower. 

.At  the  very  least,  even  if  there  are  no  im- 
mediate medical  uses  for  compounds  produced 
by  genetic  engineering,  their  indirect  impact  on 
medical  research  is  assured.  For  the  first  time, 
almost  any  biological  phenomenon  of  medical 
interest  can  be  e.xplored  at  the  cellular  level. 
These  molecules  are  valuable  tools  for  under- 
standing the  anatomy  and  functions  of  cells. 
The  knowledge  gained  may  lead  to  the  develop- 
ment of  new  therapies  or  preventive  measures 
for  diseases. 

The  chemical  industry 

FI.NDI.NGS 

The  chemical  industry's  primary  raw  materi- 
al, petroleum,  is  now  in  limited  supply.  Coal  is 
one  appealing  alternative;  another  is  biomass,  a 
renewable  resource  composed  of  plant  and  ani- 
mal material. 

Biomass  has  been  transformed  by  fermenta- 
tion into  organic  chemicals  like  citric  acid,  etha- 
nol, and  amino  acids  for  decades.  Other  organic 
chemicals  such  as  acetone,  butanol,  and  fumaric 
acid  were  at  one  time  made  by  fermentation  un- 
til chemical  production  methods,  combined 
with  cheap  oil  and  gas,  proved  to  be  more  eco- 
nomical. In  theory,  most  any  industrial  organic 
chemical  can  be  produced  by  a biological  proc- 
ess. 

Commercial  fermentation  using  genetically 
engineered  micro-organisms  offers  several  ad- 


vantages over  current  chemical  production 
technic|ues. 

• The  use  of  renewable  resources:  stai’ches, 
sugars,  cellulose,  and  other  components  of 
biomass  can  serve  as  the  raw  material  for 
synthesizing  organic  chemicals.  With  prop- 
er agricultural  management,  biomass  can 
assui'e  a continuous  renewable  supply  for 
the  industry. 

• The  use  of  physically  milder  conditions: 
chemical  processes  often  reciuire  high  tem- 
peratures  and  extreme  pressures.  These 
conditions  are  energy  intensive  and  pose  a 
hazai'd  in  case  of  accidents.  Biological  proc- 
esses operate  under  milder  conditions, 
which  are  compatible  with  living  systems. 

• One-step  production  methods:  micro-orga- 
nisms can  carry  out  several  steps  in  a syn- 
thetic process,  eliminating  the  need  for  in- 
termediate steps  of  separation  and  puri- 
fication. 

• Decreased  pollution:  because  biological 

processes  are  highly  specific  in  the  reac- 
tions they  catalyze,  they  offer  control  over 
the  products  formed  and  decrease  undesir- 
able side-products.  As  a result,  they  pro- 
duce fewer  pollutants  that  require  manage- 
ment and  disposal. 

The  impact  of  this  technology  will  cut  across 
the  entire  spectrum  of  chemical  groups:  plastics 
and  resin  materials,  flavors  and  perfumes  mate- 
rials, synthetic  rubber,  medicinal  chemicals, 
pesticides,  and  the  primary  products  from  pe- 
troleum that  serve  as  the  raw  materials  for  the 
synthesis  of  organic  chemicals.  Nevertheless, 
the  specific  products  that  will  be  affected  in 
each  group  can  only  be  chosen  on  a case-by-case 
basis,  with  the  applicability  of  genetics  de- 
pending on  a variety  of  factors.  Crude  estimates 
of  the  expected  economic  impacts  are  in  the  bil- 
lions of  dollars  per  year  for  dozens  of  chemicals 
within  20  years. 

INDUSTRY  AND  MANPOWER  IMPACTS 

Although  genetic  engineering  will  develop 
new  techniques  for  synthesizing  many  sub- 
stances, the  direct  displacement  of  any  current 
industry  seems  doubtful.  Genetic  engineering 
should  be  considered  simply  another  industrial 
tool.  Industries  will  probably  use  genetic 


8 • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


engineering  to  maintain  their  positions  in  their 
respective  markets.  This  is  already  illustrated 
hy  the  variety  of  companies  in  the  pharmaceu- 
tical, chemical,  and  energy  industries  that  have 
invested  in  or  contracted  with  genetic  engineer- 
ing firms.  Some  large  companies  are  already  de- 
veloping inhouse  genetic  engineering  research 
capabilities. 

Any  predictions  of  the  number  of  workers 
that  will  be  required  in  the  production  phase  of 
biotechnology  will  depend  on  the  expected 
volume  of  chemicals  that  will  be  produced.  At 
present,  this  figure  is  unknown.  An  estimated 
$15  billion  worth  of  chemicals  may  be  manufac- 
tured by  biological  processes.  This  will  employ 
approximately  30,000  to  75,000  workers  for  su- 
pervision, services,  and  production.  Whether 
this  will  represent  a net  loss  or  gain  in  the  num- 
ber of  jobs  is  difficult  to  predict  since  new  jobs 
in  biotechnology  will  probably  displace  some  of 
those  in  traditional  chemical  production. 

Food  processing  industry 
FINDINGS 

Genetics  in  the  food  processing  industry  can 
be  used  in  two  ways:  to  design  micro-organisms 
that  transform  inedible  biomass  into  food  for 
human  consumption  or  into  feed  for  animals; 
and  to  design  organisms  that  aid  in  food  proc- 
essing, either  by  acting  directly  on  the  food 
itself  or  by  providing  materials  which  can  be 
added  to  food. 

The  use  of  genetics  to  design  organisms  with 
desired  properties  for  food  processing  is  an 
established  practice.  Fermented  foods  and 
beverages  have  been  made  by  selected  strains 
of  mutant  organisms  (e.g.,  yeasts)  for  centuries. 
Only  recently,  however,  have  molecular  tech- 
nologies opened  up  new  possibilities.  In  par- 
ticular, large-scale  availability  of  enzymes  will 
play  an  increasing  role  in  food  processing. 

The  applications  of  molecular  genetics  are 
likely  to  appear  in  the  food  processing  industry 
in  piecemeal  fashion: 

• Inedible  biomass,  human  and  animal 
wastes,  and  even  various  industrial  efflu- 
ents are  now  being  transformed  into  edible 


micro-organisms  high  in  protein  content 
(called  single-cell  protein  or  SCP).  Its  pres- 
ent cost  of  production  in  the  United  States 
is  relatively  high,  and  it  must  compete  with 
cheaper  sources  of  protein  such  as  soy- 
beans and  fishmeal,  among  others. 

• Isolated  successes  can  be  anticipated  for 
the  production  of  such  food  additives  as 
fructose  (a  sugar)  and  the  synthetic  sweet- 
ener aspartame,  and  foi'  improxements  in 
SCP  production. 

An  industrywide  impact  is  not  expected  in  the 
near  future  because  of  several  major  conflicting 
factors: 

• The  basic  knowledge  of  the  genetic  charac- 
teristics that  could  improve  food  has  not 
been  adequately  developed. 

• The  food  processing  industry  is  conserva- 
tive in  its  expenditures  for  R&.D  to  impi  ove 
processes.  Generally,  only  one-third  to  one- 
half  as  much  is  allocated  for  this  purpose  as 
in  technologically  intensive  industries. 

• Products  made  by  new  microbial  soui'ces 
must  satisfy  the  Food  and  Drug  .Adminis- 
tration's (FDA)  safety  regulations,  which  in- 
clude undergoing  tests  to  pro\i?  lack  of 
harmful  effects.  It  may  be  possihU*  to 
reduce  the  amount  of  recjuired  testing  by 
transferring  the  desired  gejie  into  mici’o- 
organisms  that  already  meet  FDA  stand- 
ards. 

The  use  of  genetically  engineered 
micro-organisms  in  the  environment 

FINDINGS 

Genetically  engineered  micro-organisms  arc 
being  designed  now  to  p(M'torm  in  three  areas 
(aside  from  agricultui’al  u.ses)  that  r('(|uirc  their 
large-scale  release  into  th(?  (mu  ironment: 

• mineral  leaching  and  i’(h:o\  (m  v, 

• enhanced  oil  recovery,  and 

• pollution  control. 

All  of  these  are  characterized  by: 

• the  use  of  large  volumes  of  micro-orga- 
nisms, 

• decreased  control  o\ei'  the  hehaxior  .ind 
fate  of  the  micro-organisms, 


Ch.  1 — Summary:  Issues  and  Options  • 9 


• the  possibility  of  ecological  disruption,  and 

• less  de\  elopment  in  basic  R&.D  (and  more 
speculation)  than  in  the  industries  in  which 
micro-organisms  are  used  in  a controlled 
enxironment. 

Ml.VEH  XL  LEACHING  AND  RECOV  ERX 

Bacteria  ha\  e been  used  to  leach  metals,  such 
as  uranium  and  copper,  from  low-grade  ores. 
•Although  there  is  reason  to  heliexe  leaching 
ability  is  under  genetic  control  in  these  orga- 
nisms, practically  nothing  is  known  about  the 
precise  mechanisms  inxoKed.  Iherefore,  the 
application  of  genetic  technologies  in  this  area 
remains  speculatixe.  Progress  has  been  slow  in 
obtaining  more  information,  partly  because 
\ ery  little  research  has  been  conducted. 

In  addition  to  leaching,  micro-organisms  can 
be  used  to  recover  valuable  metals  or  eliminate 
polluting  metals  from  dilute  solutions  such  as  in- 
dustrial waste  streams.  The  process  makes  use 
of  the  ability  of  micro-organisms  to  bind  metals 
to  their  surfaces  and  then  concentrate  them  in- 
ternally. 

The  economic  competitiveness  of  biological 
methods  is  still  unproved,  but  genetic  modifica- 
tions have  been  attempted  only  recently.  The 
cost  of  producing  the  micro-organisms  has  been 
a major  consideration.  If  it  can  be  reduced,  the 
approach  might  be  useful. 

ENHA.NCED  OIL  RECOVERY 

Many  methods  have  been  tried  in  efforts  to 
remove  oil  from  the  ground  when  natural 
e.xpulsive  forces  alone  are  no  longer  effective. 
Injecting  chemicals  into  a reservoir  has,  in  many 
cases,  aided  recovery  by  changing  the  oil’s  flow 
characteristics. 

Micro-organisms  can  produce  the  necessary 
chemicals  that  help  to  increase  flow.  Theoreti- 
cally, they  can  also  be  grown  in  the  wells 
themselves,  producing  those  same  chemicals  in 
situ.  The  currently  favored  chemical,  xanthan, 
is  far  from  ideal  for  increasing  flow.  Genetic 
engineering  should  be  able  to  produce  chem- 
icals with  more  useful  characteristics. 

The  current  research  approach,  funded  by 
the  Department  of  Energy  (DOE)  and  independ- 
ently by  various  oil  companies,  is  a two-phase 


process  to  find  micro-organisms  that  can  func- 
tion in  an  oil  reserv  oir  en\  ironment,  and  then  to 
improve  their  chai'acteristics  genetically. 

The  genetic  alteration  of  micro-organisms  to 
produce  chemicals  useful  for  enhanced  oil  re- 
covery has  been  more  successful  than  the  alter- 
ation of  micro-organisms  that  may  be  used  in 
situ.  However,  rDNA  technology  has  not  been 
ap})lied  to  either  case.  All  attempts  have  em- 
ployed artificially  induced  or  naturally  occur- 
ring mutations. 

POLLUTION  CXINTROL 

Many  micro-organisms  can  consume  various 
kinds  of  pollutants,  changing  them  into  relative- 
ly harmless  materials  before  they  die.  These 
micro-organisms  always  have  had  a role  in 
"natural”  pollution  control:  nevertheless,  cities 
have  resisted  adding  microbes  to  their  sewerage 
systems.  Although  the  Environmental  Protec- 
tion Agency  (EPA)  has  not  recommended  addi- 
tion of  bacteria  to  municipal  sewerage  systems, 
it  suggests  that  they  might  be  useful  in  smaller 
installations  and  for  specific  problems  in  large 
systems.  In  major  marine  spills,  the  bacteria, 
yeast,  and  fungi  already  present  in  the  water 
participate  in  degradation.  The  usefulness  of 
added  microbes  has  not  been  demonstrated. 

Nevertheless,  in  1978,  the  estimated  market 
of  biological  products  for  pollution  control  was 
$2  million  to  S4  million/year,  divided  among 
some  20  companies;  the  potential  market  was 
estimated  to  be  as  much  as  $200  million/year. 

To  date,  genetically  engineered  strains  have 
not  been  applied  to  pollution  problems.  Restrict- 
ing factors  include  the  problems  of  liability  in 
the  event  of  health,  economic,  or  environmental 
damage;  the  contention  that  added  organisms 
are  not  likely  to  be  a significant  improvement; 
and  the  assumption  that  selling  microbes  rather 
than  products  or  processes  is  not  likely  to  be 
profitable. 

Convincing  evidence  that  microbes  could  re- 
move or  degrade  an  intractable  pollutant  would 
encourage  their  application.  In  the  meantime, 
however,  these  restrictions  have  acted  to  inhibit 
the  research  necessary  to  produce  marked  im- 
provements. 


10  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


CONSTRAINTS  IN  USING  GENETIC  ENGINEERING 
TECHNOLOGIES  IN  OPEN  ENVIRONMENTS 

The  genetic  data  base  for  the  potentially  use- 
ful micro-organisms  is  lacking.  Only  the  sim- 
plest methods  of  mutation  and  selection  for  de- 
sirable properties  have  been  used  thus  far. 
These  are  the  only  avenues  for  improvement 
until  more  is  learned  about  the  genetic  mech- 
anisms. 

Even  when  the  scientific  knowledge  is  avail- 
able, two  other  obstacles  to  the  use  of  geneti- 
cally engineered  micro-organisms  will  remain. 
The  first  is  the  need  to  develop  engineered 


systems  on  a scale  large  enough  to  exploit  their 
biological  activity.  This  will  necessitate  a con- 
tinual dialog  among  microbial  geneticists,  geolo- 
gists, chemists,  and  engineers;  an  interdisci- 
plinary approach  is  required  that  recognizes  the 
needs  and  limitations  of  each  discipline. 

The  second  obstacle  is  ecological.  Introducing 
large  numbers  of  genetically  engineered  micro- 
organisms into  the  environment  might  lead  to 
ecological  disruption  or  detrimental  effects  on 
human  health,  and  raise  questions  of  legal  lia- 
bility. 


Issue  and  Options — Biotechnology 


ISSUE:  How  can  the  Federal  Govern- 

ment promote  advances  in  bio- 
technology and  genetic  engi- 
neering? 

The  United  States  is  a leader  in  applying 
genetic  engineering  and  biotechnology  to  in- 
dustry. One  reason  is  the  long-standing  commit- 
ment by  the  Federal  Government  to  the  funding 
of  basic  biological  research;  several  decades  of 
support  for  some  of  the  most  esoteric  basic 
research  has  unexpectedly  provided  the  foun- 
dation for  a highly  useful  technology.  A second 
is  the  availability  of  venture  capital,  which  has 
allowed  the  formation  of  small,  innovative  com- 
panies that  can  build  on  the  basic  research. 

The  chief  argument /or  Government  subsidi- 
zation for  R&D  in  biotechnology  and  genetic 
engineering  is  that  Federal  help  is  needed  in 
areas  such  as  general  (generic)  research  or  high- 
ly speculative  investigations  not  now  being  de- 
veloped by  industry.  The  argument  against  the 
need  for  this  support  is  that  industry  will  devel- 
op everything  of  commercial  value  on  its  own. 

A look  at  what  industry  is  now  attempting  in- 
dicates that  sufficient  investment  capital  is 
available  to  pursue  specific  manufacturing  ob- 
jectives. Some  high-risk  areas,  however,  that 
might  be  of  interest  to  society,  such  as  pollution 
control,  may  justify  promotion  by  the  Govern- 
ment, while  other,  such  as  enhanced  oil  recov- 
ery might  might  not  be  profitable  soon  enough 
to  attract  investment  by  industry. 


OPTIONS: 

A.  Congress  could  allocate  funds  specifically  for 
genetic  engineering  and  biotechnology  H&..D  in 
the  budget  of  appropriate  agencies. 

Congress  could  promote  two  types  of  pro- 
grams in  biotechnology:  those  with  long-range 
payoffs  (basic  research),  and  those  that  industry 
is  not  willing  to  undertake  hut  that  might  he  in 
the  national  interest. 

B.  Congress  could  establish  a separate  Institute 
of  Biotechnology  as  a funding  agency. 

The  merits  of  a separate  institute  lie  in  the 
possibility  of  coordinating  a wide  range*  of  ef- 
forts, all  related  to  biotechnology.  On  the*  other 
hand,  biotechnology  and  genetic  engineering 
cover  such  a broad  range  of  dise'ipline's  that  a 
new  funding  agency  would  o\erlap  the  man- 
dates of  existing  agencies.  Furthe'rmore!,  the 
creation  of  yet  another  agency  carries  with  it  all 
the  disadvantages  of  increased  hur(!au('racv  atid 
competition  for  funds  at  the  agemw  level. 

C.  Congress  could  establish  research  centers  in 
universities  to  foster  interdisciplinary  ap- 
proaches to  biotechnology.  In  addition,  a pro- 
gram of  grants  could  be  offered  to  train  .sr/en- 
tists  in  biological  engineering. 

The  successful  use  of  biological  t(‘chni(|ues  in 
industry  depends  on  a multidisci|)linar\  .ip- 
proach  involving  biochemists,  getK'liiisls,  mi- 
crobiologists, process  engineers,  and  chemist s 


Ch.1 — Summary:  Issues  and  Options  *11 


Little  is  now  being  done  publicly  or  pri\  ately  to 
de\  elop  the  expertise  necessary. 

D.  Congress  could  use  ta\  incentives  to  stimulate 
hiotechnologv. 

The  tax  laws  could  be  used  to  stimulate  bio- 
technolog\'  by  expanding  the  supply  of  capital 
for  small,  high-risk  firms,  which  are  generally 
considered  more  inno\ati\e  than  established 
firms  because  of  their  w illingness  to  undertake 
the  risks  of  innoxation.  In  addition  to  focusing 
on  the  supply  of  capital,  tax  policy  could  at- 
tempt to  directly  increase  the  profitability  of 
potential  growth  companies. 

A tax  incentixe  could  also  be  directed  at  in- 
creasing R&.D  expenditures.  It  has  been  sug- 
gested that  companies  be  permitted  to  take  tax 
credits:  1)  on  a certain  percentage  of  their  R&.D 
expenses:  and  2)  on  contributions  to  unix  ersities 
! for  research. 

’ E.  Congress  could  improve  the  conditions  under 
which  U.S.  companies  collaborate  with  aca- 
demic scientists  and  make  use  of  the  technol- 
ogy developed  in  universities,  which  has  been 
wholly  or  partly  supported  by  ta\  funds. 

Dexelopments  in  genetic  engineering  have 
kindled  interest  in  this  option.  Under  legislation 
that  has  recently  passed  both  Houses  of  Con- 


gress, small  businesses  and  unix  ersities  may  re- 
tain title  to  inx  entions  developed  under  federal- 
ly funded  research.  Currently,  some  Federal 
agencies  axvard  contractors  these  exclusive 
rights,  xvhile  others  insist  on  the  nonexclusive 
licensing  of  inx  entions. 

F.  Congress  could  mandate  support  for  specific 
research  tasks  such  as  pollution  control  using 
microbes. 

Microbes  may  he  useful  in  degrading  intrac- 
table xvastes  and  pollutants.  Current  research, 
hoxvex  er,  is  limited  to  isolating  organisms  from 
natural  sources  or  from  mutated  cultures.  More 
elaborate  efforts,  involving  rDNA  techniques  or 
other  forms  of  microbial  genetic  exchange,  will 
require  additional  funding. 

G.  Most  efforts  could  be  left  to  industry  and  each 
Government  agency  allowed  to  develop  pro- 
grams in  the  fields  of  genetic  engineering  and 
biotechnology  as  it  sees  fit. 

Generic  research  xvill  probably  not  be  under- 
taken by  any  one  company.  Leaving  all  R&,D  in 
industry’s  hands  would  still  produce  major  com- 
mercial successes,  but  does  not  ensure  the  de- 
x elopment  of  needed  basic  general  knowledge 
or  the  undertaking  of  high-risk  projects. 


Agriculture  

The  complexity  of  plants  and  animals  pre- 
sents a greater  challenge  to  advances  in  applied 
genetics  than  that  posed  by  micro-organisms. 
Nexertheless,  the  successful  genetic  manipula- 
tion of  microbes  has  encouraged  researchers  in 
the  agricultural  sciences.  The  nexv  tools  xvill  be 
used  to  complement,  but  not  replace,  the  well- 
established  practices  of  plant  and  animal 
breeding. 

The  applications  of  genetics  to  plants 

FIXDIIVGS 

It  is  impossible  to  exactly  determine  the  ex- 
tent to  xvhich  applied  genetics  has  directly  con- 
tributed to  increases  in  plant  yield  because  of 
simultaneous  improvements  in  farm  manage- 


ment, pest  control,  and  cropping  techniques 
using  herbicides,  irrigation,  and  fertilizers. 
Nevertheless,  the  impacts  of  breeding  technol- 
ogies have  been  extensive. 

The  plant  breeder’s  approach  is  determined 
for  the  most  part  by  the  particular  biological 
factors  of  the  crop  being  bred.  The  new  genetic 
technologies  potentially  offer  additional  tools  to 
allow  development  of  new  varieties  and  even 
species  of  plants  by  circumventing  current  bio- 
logical barriers  to  the  exchange  of  genetic 
material. 

Technologies  developed  for  classical  plant 
breeding  and  those  of  the  new  genetics  should 
not  be  viewed  as  being  compretftive;  they  are 
both  tools  for  effectively  manipulating  genetic 


12  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


information.  One  new  technology— e.g.,  proto- 
plast fusion,  or  the  artificial  fusion  of  two  cells— 
allows  breeders  to  overcome  incompatibility 
between  plants.  But  the  plant  that  may  result 
still  must  be  selected,  regenerated,  and  eval- 
uated under  field  conditions  to  ensure  that  the 
genetic  change  is  stable  and  that  the  attributes 
of  the  new  variety  meet  commercial  require- 
ments. 

In  theory,  the  new  technologies  will  expand 
the  capability  of  breeders  to  exchange  genetic 
information  by  overcoming  natural  breeding 
barriers.  To  date,  however,  they  have  not  had  a 
widespread  impact  on  the  agricultural  industry. 

As  a note  of  caution,  it  must  be  emphasized 
that  no  plant  can  possess  every  desirable  trait. 
There  will  always  have  to  be  some  tradeoff; 


A young  Douglas  fir  tree  propagated  4 years  ago  (rom  a 
small  piece  of  seedling  leaf  tissue.  Three  years  ago  this  v.as 
at  the  test-tube  stage  seen  in  the  loblolly  pine  photograph 


often  quality  for  quantity,  such  as  increased 
protein  content  but  decreased  yield. 

NEW  GENETIC  TECHNOLOGIES  FOR 
PLANT  BREEDING 

The  new  technologies  fall  into  two  categories: 
those  involving  genetic  transformations 
through  cell  fusion  and  those  involving  the  in- 
sertion or  modification  of  genetic  information 
through  the  cloning  of  DNA  and  its  vectors. 
Techniques  are  available  for  manipulating 
organs,  tissues,  cells,  or  protoplasts  in  culture; 
for  regenerating  plants;  and  for  testing  the 
genetic  basis  of  novel  traits.  So  far  these  tech- 
niques are  routine  only  in  a few  species. 

The  approach  to  exploiting  molecular  biology 
for  plant  breeding  is  similar  in  some  respects  to 
the  genetic  manipulation  of  micro-organisms. 
However,  there  is  one  major  conceptual  dif- 


Photo  Cftdif:-  -r'Co 


A plantlet  of  loblolly  pine  grown  in  Weyerhaeuser  Co.’s 
tissue  culture  laboratory.  The  next  step  in  this  procedure 
is  to  transfer  the  plantlet  from  its  sterile  and  humid 
environment  to  the  soil 


c/7.  1— Summary:  Issues  and  Options  • 13 


I ferenre.  In  miri'o-organisms.  the  dianges  madt' 
j on  tlie  cellular  le\el  are  the  goals  ot  the 
I manipulation.  With  ci'ops,  changes  made  on  the 
i cellular  le\  el  art'  meaningless  unless  they  can  he 
I reproduced  in  tiie  entirt'  plant  as  well.  There- 
! fore,  unless  single  cells  in  culture  can  he 
selected  atul  grow  n into  mature  plants  and  the 
desired  traits  e.xpressed  in  the  mature  plant— 
procedures  w hich  at  this  lime  ha\  e had  limited 
success— the  benefits  of  genetic  engineei’ing  w ill 
i not  he  w idely  felt  in  plant  hreeding. 

.Moderate  success  has  been  achie\ed  for 
growing  cells  in  tissiu'  cultui'e  into  matui'e 
plants.  I'issue  culture  programs  of  commercial 
significance  in  the  I'nited  States  include  the 
asparagus,  citrus  fruits,  pineapples,  and  straw- 
berries. Breeders  ha\e  had  little  success,  how- 
e\er,  in  regenerating  mature  plants  of  wide 
agronomic  impoi  tance,  such  as  corn  and  w heat. 

Some  success  can  he  claimeil  for  engineering 
changes  to  alter  genetic  makeup.  Both  the  stable 
integration  of  genetic  material  into  a cell  and 
the  fusion  of  genetically  different  cells  are  still 
largely  e.xperimenlal  techniques.  Technical 
i breakthroughs  ha\e  come  on  a species-hy- 
! species  basis,  hut  ke\'  disco\  eries  are  not  often 
I applicable  to  all  plants.  Initial  results  suggest 
I that  agronomically  important  traits,  such  as 
disease  resistance,  can  be  transferred  from  one 
I species  to  another.  Limited  success  has  also 
I been  shown  in  attempts  to  create  totally  new 
species  by  fusing  cells  from  different  genera. 
.Attempts  to  find  both  suitable  \ ectors  and  genes 
for  transferring  one  plant  s genes  to  another  are 
only  now  beginning  to  show  promise. 

CO\STRAI.\TS  0.\  L'SLNG  MOLECULAR 
GENETICS  FOR  PLANT  IMPRO\'EMENTS 

Molecular  engineering  has  been  impeded  by  a 
lack  of  answers  to  basic  questions  in  molecular 
biolog\'  and  plant  physiology'  owing  to  insuffi- 
cient research.  Federal  funding  for  plant  molec- 
ular genetics  in  agriculture  has  come  primarily 
from  the  U.S.  Department  of  Agriculture 
(USDA)  and  the  National  Science  Foundation 
(NSF).  In  USDA,  research  support  is  channeled 
primarily  through  the  flexible  competitive 
grants  program  (fiscal  year  1980  budget  of  $15 
million)  for  the  support  of  new’  research  direc- 
bons  in  plant  biology.  The  total  support  for  the 


plant  sciences  from  NSF  is  approximately  $25 
million,  only  $1  million  of  which  is  specifically 
designated  for  plant  genetics. 

rhe  shortage  of  a trained  workforce  is  a 
significant  constraint.  Only  a few  universities 
have  expertise  in  both  plants  and  molecular  bi- 
ology. In  addition,  there  are  only  a few  people 
w ho  have  the  ability  to  work  with  modern  mo- 
leculai’  techni(|ues  related  to  whole  plant  prob- 
lems. ,\s  a result,  a business  firm  could  easily 
de\elo|)  a capability  in  this  area  exceeding  that 
at  any  indi\  idual  U.S.  university.  However,  the 
building  of  industrial  laboratories  and  suhse- 
c|uent  hiring  from  the  universities  could  easily 
cleplete  the  expertise  at  the  university  level. 
With  the  recent  investment  activity  by  many 
bioengineering  firms,  this  trend  has  already 
begun;  in  the  long-run  it  could  have  serious  con- 
sequences for  the  ciualitv  and  quantity  of  uni- 
versity research. 

GENETIC  VARIABILITY,  CROP  VULNERABILITY, 
AM)  THE  STORAGE  OF  GERMPLASM 

Successful  plant  breeding  is  based  on  tbe 
availability  of  genetically  diverse  plants  for  the 
insertion  of  new  genes  into  plants.  The  number 
of  these  plants  has  been  diminishing  for  a varie- 
ty of  reasons.  However,  the  rate  and  extent  of 
this  trend  is  unknown;  the  data  simply  do  not 
exist.  Therefore,  it  is  essential  to  have  an  ade- 
quate scientific  understanding  of  how'  much  ge- 
netic loss  has  taken  place  and  how^  much  germ- 
plasm  (the  total  genetic  v'ariability  available  to  a 
species)  is  needed.  Neither  of  these  questions 
can  be  answered  completely  at  this  time. 

Even  if  genetic  needs  can  be  adequately  iden- 
tified, there  is  disagreement  about  the  quantity 
of  germplasm  to  collect.  Furthermore,  the  ex- 
tent to  which  the  new  genetic  technologies  will 
affect  genetic  variability,  vulnerability,  or  the 
storage  technologies  of  germplasm  has  not  been 
determined.  As  a result,  it  is  currently  difficult, 
if  not  impossible,  to  state  how  much  effort 
should  be  expended  by  the  National  Germplasm 
System  to  collect,  maintain,  and  test  new  gene 
resources  (in  this  case  as  seed). 

Finally,  even  if  an  adequate  level  of  genetic 
variability  can  be  assessed,  the  real  problem  of 
vulnerability— the  practice  of  planting  only  a 


14  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


single  variety— must  be  dealt  with  at  an  institu-  nologies  existed,  farmers  would  still  select  only 

tional  or  social  level.  Even  if  no  genetic  tech-  one  or  a few  “best”  varieties  for  planting. 


ISSUE:  Should  an  assessment  be  con- 

ducted to  determine  how  much 
diversity  in  plant  germplasm 
needs  to  be  maintained? 

An  understanding  of  how  much  germplasm 
should  be  protected  and  maintained  would 
make  the  management  of  genetic  resources 
simpler. 

OPTIONS: 

A.  Congress  could  commission  a study  of  how 
much  genetic  variability  is  necessary  or  desir- 
able to  meet  present  and  future  needs. 

A comprehensive  evaluation  of  the  National 
Germplasm  System’s  requirements  for  collec- 
ting, evaluating,  maintaining,  and  distributing 
genetic  resources  for  plant  breeding  and  re- 
search could  serve  as  a baseline  for  a further 
assessment. 

B.  Congress  could  commission  a study  on  the 
need  for  international  cooperation  to  manage 
and  preserve  genetic  resources  both  in  natural 
ecosystems  and  in  repositories. 

This  investigation  could  include  an  evaluation 
of  the  rate  at  which  genetic  diversity  is  being 
lost  from  natural  and  agricultural  systems  along 
with  an  estimate  of  the  effects  this  loss  will 
have. 

C.  Congress  could  commission  a study  on  how  to 
develop  an  early  warning  system  to  recognize 
the  potential  vulnerability  of  crops. 

Where  high  genetic  uniformity  still  exists, 
proposals  could  be  suggested  to  reduce  any 
risks  due  to  uniformity.  Alternatively,  the 
avenues  by  which  private  seed  companies  could 
be  encouraged  to  increase  the  levels  of  genetic 
diversity  could  be  investigated. 


What  are  the  most  appropriate 
approaches  in  overcoming  the 
various  technical  constraints 
that  limit  the  success  of  molec- 
ular genetics  for  plant  improve- 
ment? 

Although  genetic  information  has  been  trans- 
ferred by  vectors  and  protoplast  fusion,  1)N,\ 
transformations  of  commercial  \ alue  ha\  e not 
yet  been  performed.  Molecular  engineering  has 
been  impeded  by  the  lack  of  vectors  that  can 
transfer  novel  genetic  material  into  plants, 
by  insufficient  knowledge  about  which  genes 
would  be  useful  for  breeding  purposes,  and  by 
a lack  of  understanding  of  the  incompatibility  of 
chromosomes  from  diverse  sources.  ,\noth(M' 
impediment  has  been  the  lack  of  researc'hei's 
from  a variety  of  disciplines. 

OPTIONS: 

A.  The  level  of  funding  could  be  increased  for 
plant  molecular  genetics  research  supfiorled 
by  NSF  and  USDA. 

B.  Research  units  devoted  to  plant  molecular  ge- 
netics could  be  established  under  the  auspices 
of  the  National  Institutes  of  Health  (NIW,  with 
emphasis  on  potential  pharmaceuticals  de- 
rived from  plants. 

C.  An  institute  for  plant  molecular  genetics  could 
be  established  under  the  Science  and  educa- 
tion Administration  at  LLSDA  that  would  in- 
clude multidisciplinary  teams  to  consider  both 
basic  research  questions  and  direct  applica- 
tions of  the  technologv  to  commercial  net'ds 
and  practices. 

The  discoveries  of  mokunilar  |)lant  genetics 
will  be  used  in  conjunction  with  traditional 
breeding  programs.  Henct?,  (>ach  of  the  ihia'c 
options  could  retiuire  additional  ap|)ro[)riaiions 
for  agricultural  research. 


Issues  and  Options — Plants 
ISSUE: 


Atli  itncvs  in  repriuliictive  biolo^  nrul 
their  i'Jf'ei'ts  an  nninuil  improvement 

I i\i)i\(;s 

Much  impi'()\  ement  can  he  made  in  the  ^erm- 
plasm  of' all  major  farm  animal  species  using  ex- 
isting technologN . The  twpancled  use  of  artificial 
insemination  ( \l)  with  stored  frozen  s|)(M'm,  es- 
pecialK  in  heef  cattle,  would  benefit  both  pro- 
ducers and  consumers.  New  technicjues  for  syn- 
chronizing estrus  should  encourage  the  wider 
use  of  W.  \ arious  manipulations  of  embryos 
will  find  limited  use  in  })roducing  breeding 
stocks,  and  sex  selection  anil  twinning  tech- 
niques should  he  available  for  limiteii  applica- 
tions w ithin  the  next  10  to  20  years. 

rhe  most  important  technologv'  in  reproduc- 
the  physiology  will  continue  to  he  .\l.  Due  in 
part  to  genetic  improvement,  the  average  milk 
yield  of  cows  in  the  United  States  has  more  than 
doubled  in  the  past  30  years,  while  the  total 
number  of  milk  cow  s has  been  reduced  by  more 
than  half.  .AI  along  w ith  improv  ed  management 
and  the  av  ailahility  and  use  of  accurate  progeny 
records  on  breeding  stock  have  caused  this 
great  increase.  (See  figure  3.) 

The  improvement  lags  behind  what  is  theo- 
retically possible.  In  practice,  the  observed  in- 
j crease  is  about  100  lb  of  milk  per  cow  per  year, 

> while  a hypothetical  breeding  program  using  Al 
would  result  in  a yearly  gain  of  220  lb  of  milk 
per  cow.  The  biological  limits  to  this  rate  of  gain 
are  not  known. 

In  comparison  w ith  dairy  cattle,  the  beef  cat- 
tle industry  bas  not  applied  .AI  technology'  wide- 
ly. Only  3 to  5 percent  of  U.S.  beef  is  artificially 
inseminated,  compared  to  60  percent  of  the 
dairy  herd.  This  low  rate  for  beef  cattle  can  be 
i explained  by  sev  eral  factors,  including  manage- 
ment techniques  (range  v.  confined  housing) 
and  the  conflicting  objectives  of  individual 
breeders,  ranchers,  breed  associations,  and 
commercial  farmers. 

The  national  calf  crop— calves  aliv  e at  vv  ean- 
ing  as  a fraction  of  the  total  number  of  cows  ex- 
posed to  breeding  each  year— is  only  65  to  81 
percent.  An  improv'ement  of  only  a few  percent- 
age points  through  AI  would  result  in  savings  of 


C/7.  7 — Summary:  Issues  and  Options  • 15 


hundreds  of  millions  of  dollars  (o  proilucers  and 
consumers. 

tx)upled  with  a technology  for  estrus-cycle 
regulation,  the  use  of  ,Al  could  he  expanded  for 
both  dairy  and  heef  breeding.  Kmbi’vo  ti'ansfer 
technology,  ali'eady  well-developed  hut  still 
costly,  can  he  used  to  produce  valuable  breed- 
ing stock.  Sexing  technology,  which  is  not  yet 
perfected,  would  he  of  enormous  benefit  to  the 
beef  inilustry  because  bulls  grow  faster  than 
heifers. 

In  the  case  of  animals  other  than  cows: 

• K.\[)anded  use  of  AI  for  swine  proiluction 
will  he  encouraged  by  the  stiong  trend  to 
confinement  housing,  although  the  poor 
ability  of  boar  sperm  to  withstand  freezing 
vv  ill  continue  to  be  a handicap. 

• The  benefits  of  applied  genetics  have  not 
been  realized  in  sheep  production  because 
neilbei'  AI  nor  performance  testing  bas 
been  used.  As  long  as  the  use  of  AI  con- 
tinues to  be  limited  by  tbe  inability  to 
freeze  semen  and  by  a lack  of  agents  on  the 
market  for  synchronizing  estrus,  no  rapid 
major  gains  can  he  expected. 

• Increasing  interest  in  goats  in  the  United 
States  and  the  demand  for  goat  products 
throughout  the  world,  should  encourage 
attention  to  the  genetic  gains  that  the  use 
of  AI  and  other  technologies  make  possible. 

• Poultry  breeders  will  continue  to  concen- 
trate on  improved  egg  production,  growth 
rate,  feed  efficiency,  and  reduced  body  fat 
and  diseases.  The  use  of  frozen  semen 
should  increase  as  will  the  use  of  AI  and 
dwarf  broiler  breeders. 

• Genetics  applied  to  production  of  fish, 
mollusks,  and  crustaceans  in  either  natural 
environments  or  manmade  culture  systems 
is  only  at  the  rudimentary  stage. 

Breeders  must  have  reliable  information 
about  the  genetic  value  of  the  germplasm  they 
are  considering  introducing.  Since  farmers  do 
not  have  the  resources  to  collect  and  process 
data  on  the  performance  of  animals  other  than 
those  in  their  owm  herds,  they  must  turn  to  out- 
side sources.  The  National  Cooperative  Dairy 
Herd  Improvement  Program  (NCDHIP)  is  a mod- 


16  • The  Impacts  of  Genetics:  Applications  to  Micro-Organisms,  Plants,  and  Animals 


Figure  3.— The  Way  the  Reproductive  Technologies  Interrelate 


Bull 


Sperm 


Superovulated 

cow 


% 


Recovered 

embryos 


Sexed? 


Photo  Credit  r-  . ’•  r 


These  10  calves  from  Colorado  State  University  were  the 
result  of  superovulation,  in  vitro  culture,  and  transfer  to 
the  surrogate  mother  cows  on  the  left.  The  genetic 
mother  of  all  10  calves  is  at  upper  right 


SOURCE:  Office  of  Technology  Assessment. 


Ch.  1— Summary:  Issues  and  Options  • 17 


el  information  system  and  coolcl  be  adapted  to 
other  species. 

Selection— deciding  which  animals  to  mate 
—is  the  breeder’s  most  basic  tool.  When  going 
outside  his  herd  to  purchase  new  germplasm, 
the  breeder  needs  im[)artial  information  about 
the  quality  of  the  a\  ailahle  germplasm.  \CDHlP 
had  recorded  2.8  million  of  the  10.8  million  U.S. 
dairy  cattle  in  1979.  In  1978,  cows  enrolled  in 
the  official  plans  of  NCDHIP  outproduced  cow  s 
not  enrolled  by  vT.OOO  Ih  of  milk  per  cow , re[)re- 
senting  52  percent  more  milk  per  lactation. 

\o  comparable  information  system  e.xists  for 
other  types  of  li\  estock.  Beef  hulls,  for  e.xample, 
continue  to  be  sold  to  a large  extent  on  the  basis 
of  pedigrees,  hut  with  relatixely  little  objectiv  e 
information  on  their  genetic  merit.  Data  on 
dairy  goats  in  the  L’nited  States  became  avail- 
able through  \CDHIP  for  the  first  time  in  late 
1980.  No  nationwide  information  systems  exist 


foi'  other  species,  although  pork  production  in 
the  United  States  would  greatly  benefit  from  a 
national  swine  testing  program. 

The  more  esotei'ic  methods  of  genetic  manip- 
ulation will  probably  have  little  impact  on  the 
production  of  animals  or  animal  products  with- 
in the  tiext  10  years.  Other  in  vitro  manij)ula- 
tions,  such  as  cloning,  cell  fusion,  the  produc- 
tion of  chimeras,  and  the  use  of  rDNA  tech- 
ni(|ues,  will  continue  to  he  of  intense  interest, 
especially  for  research  purposes.  It  is  less  likely, 
however,  that  they  will  have  widespread  prac- 
tical effects  on  farm  production  in  this  century. 

Each  lechni(iue  requires  more  research  and 
refinement.  Lhitil  specific  genes  of  farm  animals 
can  he  identified  and  located,  no  direct  gene 
manipulation  will  he  practicable.  In  addition 
this  will  he  difficult  because  most  traits  of  im- 
portance are  due  to  multiple  genes. 


Issue  and  Options — Animals 


ISSUE:  How  can  the  Federal  Govern- 

ment improv  e the  germplasm  of 
major  farm  animal  species? 

OPTIONS: 

A.  Programs  like  the  i\CDHIP  could  have  in- 
creased governmental  participation  and  fund- 
ing. The  efforts  of  the  Beef  Cattle  Improve- 
ment Federation  to  standardize  procedures 
could  receive  active  support,  and  a similar  in- 
formation system  for  swine  could  be  estab- 
lished. 

The  fastest  and  least  expensive  way  to  up- 
grade breeding  stock  in  the  United  States  is 
through  effective  use  of  information.  Computer 
technologv,  along  with  a network  of  local  repre- 

The  wide  variety  of  applications  for  genetic 
engineering  is  summarized  in  figure  4.  Genetics 
can  be  used  to  improve  or  increase  the  quality 
and  output  of  plants  and  animals  for  direct  use 
by  man.  Alternatively,  materials  can  be  ex- 
tracted from  plants  and  animals  for  use  in  food, 
chemical,  and  pharmaceutical  industries. 


sentatives  for  data  collecting,  can  provide  the 
indiv  idual  farmer  or  breeder  with  accurate  in- 
formation on  the  available  germplasm  so  that  he 
can  make  his  own  breeding  decisions. 

This  option  implies  that  the  Federal  Govern- 
ment would  play  such  a role  in  new  programs, 
and  expand  its  role  in  existing  ones. 

B.  Federal  funding  could  be  increased  for  basic 
research  in  total  animal  improvement. 

This  option,  in  contrast  to  option  A,  assumes 
that  it  is  necessary  to  maintain  or  expand  basic 
R&D  to  generate  new  knowledge  that  can  be 
applied  to  the  production  of  improved  animals 
and  animal  products. 

♦ 

Biological  materials  can  also  be  converted  to 
useful  products.  In  this  process,  genetic  engi- 
neering can  be  used  to  develop  micro-organisms 
that  will  carry  out  the  conversions.  Therefore, 
genetic  manipulation  cannot  only  provide  more 
or  better  biological  raw  materials  but  can  also 
aid  in  their  conversion  to  useful  products. 


18  • The  Impacts  of  Genetics:  Applications  to  Micro-Organisms,  Plants,  and  Animals 


Figure  4.— Applications  of  Genetics 


AGRICULTURAL 

INDUSTRY 


Plants 


* t 

Animals 


Genetics 


I [ 


Direct  use 
as  food 


Convert  to 
food 


(Increase/Improve  Output) 


f 


Direct  extraction  of 
chemicals 


Production  of 
chemicals 


Direct  extraction  of 
pharmaceuticals 


Production  of 
pharmaceuticals 


Micro-organisms 


Genetics 


Micro-organisms 


FOOD 

INDUSTRY 


Micro-organisms 
{Genetics 


Micro-organisms 


CHEMICAL 

INDUSTRY 


Micro-organisms 


Genetics 


Micro-organisms 


PHARMACEUTICAL 

INDUSTRY 


SOURCE:  Office  of  Technology  Assessment. 


Institutions  and  society 

Regulation  of  genetic  engineering 
FINDINGS 

No  evidence  exists  that  any  unexpectedly 
harmful  genetically  engineered  organism  has 
been  created.  Yet  few  experts  believe  that  mo- 
lecular genetic  techniques  are  totally  without 
risk  to  health  and  the  environment.  Information 
that  has  proved  useful  in  assessing  the  risks 
from  these  techniques  has  come  from  three 
sources:  experiments  designed  specifically  to 
test  the  consequences  of  working  with  rDNA, 
experiments  designed  for  other  purposes  but 


relevant  to  rDNA,  and  scientific  meetings  and 
workshops. 

A program  of  risk  assessment  was  (fstahlished 
at  NIH  in  1979  to  conduct  exj)eriments  and  col- 
late relevant  information.  It  assesses  one  form 
of  genetic  engineering,  rDNA.  On  the  basis  of 
these  data,  conjectured,  inadvertant  risk  is 
generally  regarded  as  less  likely  today  than 
originally  suspected.  Risk  due  to  the  mani[)ula- 
tion  of  genes  from  organisms  known  to  he  haz- 
ardous is  considered  to  he  more  realistic.  T here- 
fore,  microbiological  safety  precautions  that  are 


Ch.  1— Summary:  Issues  and  Options  • 19 


appropriate  to  the  use  of  the  micro-organisms 
serx  ing  as  the  source  of  n\A  are  reciuirecl.  Nev- 
ertheless. it  has  not  been  demonstrated  that 
comhining  those  genes  in  the  form  of  rDNA  is 
anv  more  hazardous  than  tlie  original  source  of 
the  DNA. 

Perceptions  of  the  nature,  magnitude,  and  ac- 
ceptability of  the  I’isk  differ.  In  addition,  public 
concern  has  been  e.xpressed  about  possible 
long-range  im[)acts  of  genetic  engineering.  In 
this  conte.xt,  the  problem  facing  the  policy- 
maker is  how  to  address  the  risk  in  a way  that 
accommodates  the  perceptions  and  \alues  of 
those  who  hear  it. 

The  N'lH  (iuidelines  for  Research  Inxohing 
Recombinant  DN'.A  .Molecules  and  existing  Fed- 
eral laws  appear  adequate  in  most  cases  to  deal 
v\ith  the  risks  to  health  and  the  enxironment 
presented  by  genetic  engineering.  Howex  er,  the 
(iuidelines  are  not  legally  binding  on  industry, 
and  no  singU’  statute  oi’  combination  xx  ill  clearly 
cox  er  all  foreseeable  commercial  applications  of 
genetic  engineering. 

The  Guidelines  are  a flexible  exolx  ing  oxer- 
sight  mechanism  that  combines  technical  exper- 
tise xvith  public  participation.  They  coxer  the 
most  xvidely  used  and  possibly  risky  molecular 
genetic  technique— rDN'A— prohibiting  experi- 
ments using  dangerous  toxins  or  pathogens  and 
setting  containment  standards  for  other  poten- 
tially hazardous  experiments.  .Although  compli- 
ance is  mandatory  only  for  those  receix  ing  NIH 
funds,  other  Federal  agencies  folloxv  them,  and 
industry  has  proclaimed  voluntary  compliance. 
Rare  cases  of  noncompliance  have  occurred  in 
universities  but  have  not  posed  risks  to  health 
or  the  environment.  As  scientists  hax  e learned 
more  about  rDNA  and  molecular  genetics,  the 
restrictions  have  been  progressively  and  sub- 
stantially relaxed  to  the  point  xvhere  85  percent 
of  the  experiments  can  noxv  be  done  at  the 
loxvest  containment  levels,  and  virtually  all 
monitoring  for  compliance  noxv  rests  xvith  ap- 
proximately 200  local  self-regulatory  commit- 
tees called  institutional  biosafety  committees 
(IBCs).  (See  table  1.) 

Under  the  Guidelines,  NIH  serx'es  an  impor- 
tant oversight  role  by  sponsoring  risk  assess- 


Table  1.— Containment  Recommended  by  the 
National  Institutes  of  Health 


Biological — Any  connbination  of  vector  and  host  must  be 
chosen  to  minimize  both  the  survival  of  the  system 
outside  of  the  laboratory  and  the  transmission  of  the 
vector  to  nonlaboratory  hosts.  There  are  three  levels 
of  biological  containment: 

HV1—  Requires  the  use  of  Escherichia  coli  K12  or 

other  weakened  strains  of  micro-organisms  that 
are  less  able  to  live  outside  the  laboratory. 

HV2—  Requires  the  use  of  specially  engineered  strains 
that  are  especially  sensitive  to  ultraviolet  light, 
detergents,  and  the  absence  of  certain 
uncommon  chemical  compounds. 

HV3—  No  organism  has  yet  been  developed  that  can 
qualify  as  HV3. 

Physical — Special  laboratories  (P1-P4) 

PI—  Good  laboratory  procedures,  trained  personnel, 
wastes  decontaminated 

P2—  Biohazards  sign,  no  public  access,  autoclave  in 
building,  hand-washing  facility 

P3—  Negative  pressure,  filters  in  vacuum  line,  class  II 
safety  cabinets 

P4—  Monolithic  construction,  air  locks,  all  air 
decontaminated,  autoclave  in  room,  all 
experiments  in  class  III  safety  cabinets  (glove 
box),  shower  room 


SOURCE:  Office  of  Technology  Assessment. 

ment  programs,  certifying  nexv  host-vector  sys- 
tems, serx  ing  as  an  information  clearinghouse, 
and  coordinating  Federal  and  local  activities. 
Limitations  in  NIH’s  oversight  are  that:  it  lacks 
legal  authority  ox  er  industry;  its  procedures  for 
adx’ising  industry  on  large-scale  projects  have 
not  incorporated  sufficient  expertise  on  large- 
scale  fermentation  technology;  its  monitoring 
for  either  compliance  or  consistent  application 
of  the  Guidelines  by  individuals  or  institutions  is 
x'irtually  nonexistent;  and  it  has  not  systemati- 
cally ex  aluated  other  techniques,  such  as  cell  fu- 
sion, that  might  present  risks. 

Federal  laws  on  health  and  environment  will 
coxier  most  commercial  applications  of  genetic 
engineering.  Products  such  as  drugs,  chemicals, 
and  foods  can  be  regulated  by  existing  laws. 
However,  uncertainty  exists  about  the  regula- 
tion of  either  production  methods  using  engi- 
neered micro-organisms  or  their  intentional 
release  into  the  environment,  when  the  risk  has 
not  been  clearly  demonstrated.  While  a broad 
interpretation  of  certain  statutes,  such  as  the 
Occupational  Safety  and  Health  Act  and  the 
Toxic  Substances  Control  Act,  might  cover  these 


20  • Impacts  of  Applied  Genetics— Micro-Organisms,  Piants,  and  Animals 


situations,  regulatory  actions  based  on  such  in- 
terpretations could  be  challenged  in  court.  In 
anv  e\ent,  those  agencies  that  could  have 


substantial  regulatory  authority  over  commer- 
cial genetic  engineering  have  not  yet  officially 
acted  to  assert  that  authority. 


Issue  and  Options — Regulation 


ISSUE:  How  could  Congress  address  the 

risks  presented  by  genetic  engi- 
neering? 

OPTIONS: 

A.  Congress  could  maintain  the  status  quo  by  let- 
ting NIH  and  the  regulatory  agencies  set  the 
Federal  policy. 

Congress  might  determine  that  legislation  to 
remedy  the  limitations  in  current  Federal  over- 
sight would  result  in  unnecessary  and  burden- 
some regulation.  No  known  harm  to  health  or 
the  en\'ironment  has  occurred  under  current 
regulation.  Also  the  agencies  generally  have  the 
legal  authority  and  expertise  to  adapt  to  most 
new  problems  posed  by  genetic  engineering. 

The  disadvantages  are  the  lack  of  a central- 
ized, uniform  Federal  response  to  the  problem, 
and  the  possibility  that  risks  associated  with 
commercial  applications  will  not  be  adequately 
addressed.  Conflicting  or  redundant  regulations 
of  different  agencies  would  result  in  unneces- 
sary burdens  on  those  regulated. 

B.  Congress  could  require  that  the  Federal  Inter- 
agency Advisory  Committee  on  Recombinant 
DNA  Research  prepare  a comprehensive  re- 
port on  its  members'  collective  authority  to 
regulate  rDNA  and  on  their  regulatory  inten- 
tions. 

The  Industrial  Practices  Subcommittee  of  this 
Committee  has  been  studying  agency  authority 
over  commercial  rDNA  activities.  Presently, 
there  is  little  official  guidance  on  regulatory  re- 
quirements for  companies  that  may  soon  mar- 
ket products  made  by  rDNA  methods.  A con- 
gressionally  mandated  report  would  ensure  full 
consideration  of  these  issues  by  the  agencies 
and  expedite  the  process.  On  the  other  hand, 
the  agencies  are  studying  the  situation,  which 
must  be  done  before  they  can  act.  Also,  it  is 
often  easier  and  more  efficient  to  act  on  each 
case  as  it  arises,  rather  than  on  a hypothetical 
basis  before  the  fact. 


C.  Congress  could  require  that  all  recombinant 
DNA  activity  be  monitored  for  a limited  num- 
ber of  years. 

This  represents  a "wait  and  see"  |K)silion  by 
Congress  and  the  middle  ground  between  the 
status  quo  and  full  regulation.  It  recognizes  and 
balances  the  following  factors:  1)  the  absence  of 
demonstrated  harm  to  human  health  or  the  en- 
vironment from  genetic  engineering;  2)  the  con- 
tinuing concern  that  genetic  engineering  pre- 
sents risks;  3)  the  lack  of  sufficient  knowledge 
and  experience  from  which  to  make  a final  judg- 
ment; 4)  the  existence  of  an  oversight  mech- 
anism that  seems  to  be  working  well,  hut  that 
has  clear  limitations  with  respect  to  commercial 
activities;  5)  the  virtual  abolition  of  Federal 
monitoring  of  rDNA  acti\  ities  by  recent  amend- 
ments to  the  Guidelines;  and  6)  the  expected  in- 
crease in  commercial  genetic  engineering. 

This  option  would  pro\  ide  a data  ha.se  that 
could  be  used  for:  1)  determining  the  effec- 
tiveness of  voluntary  compliance  with  the 
Guidelines  by  industry,  and  mandatory  com[)li- 
ance  by  Federal  grantees;  2)  determining  the 
quality  and  consistency  of  the  local  self-regu- 
latory actions;  3)  continuing  a formal  risk  ass('ss- 
ment  program;  4)  identifying  \ ague  oi'  conflict- 
ing provisions  of  the  Guidelines  for  rev  ision;  .3) 
identifying  emerging  trends  or  problems;  and  (i) 
tracing  any  long-term  adverse  im[)acts  on  health 
or  the  environment  to  their  soui'ces. 

The  obvious  disadvantage  of  this  option 
would  be  the  required  paperwork  and  (dfort  by 
scientists,  universities,  corporations,  and  the 
Federal  Government. 

D.  Congress  could  make  the  NIH  Cuidelines  ap- 
plicable to  all  rDNA  work  done  in  the  I 'idled 
States. 

This  option  would  eliminate  any  concern 
about  the  effectiveness  of  voluntary  ('ompliance 
with  the  Guidelines,  and  it  has  the  ad\  antage  ot 


Ch.  1— Summary:  Issues  and  Options  • 21 


using  an  existing  o\  ersight  mechanism.  I'he  ma- 
jor changes  that  uDukl  have  to  he  made  in  the 
area  ot  enforcement.  I’rest'iit  penalties  for  non- 
compliance— suspension  or  termination  of  re- 
search funtls— ai'('  ohv  iously  inapplic  able  to  in- 
dustry. In  addition,  procedui'c's  for  monitoi'ing 
compliance'  would  ha\  e to  he  strengthenc'd. 

I'lie  main  disadvantage  of  this  option  is  that 
MH  is  not  a regulatory  agency.  Since  \'IH  has 
traditionally  viewed  its  mission  as  promoting 
biomedical  research,  it  would  have  a conflict  of 
interest  between  regulation  and  promotion. 
One  of  the  regulatory  agencies  could  he  given 
the  authority  to  enforce  the  (lUidelines. 

£.  Congress  could  require  an  environmental  im- 
pact statenient  and  agency  approval  before 
any  genetically  engineered  organism  is  inten- 
tionally released  into  the  environnwnt. 

There  have  been  numerous  cases  where  an 
animal  or  plant  species  has  been  introduced  into 
a new  env  ironment  and  has  spread  in  an  uncon- 
trolled and  undesirable  fashion.  Vet  in  pollution 
control,  mineral  leaching,  and  enhanced  oil 
recov  erv,  it  might  be  desirable  to  release  large 
numbers  of  engineered  micro-organisms  into 
the  environment. 

The  Guidelines  currently  prohibit  deliberate 
release  of  any  organism  containing  rDNA  with- 
out approval  of  NIH.  One  disadvantage  of  this 
prohibition  is  that  it  lacks  the  force  of  law. 
.Another  is  that  approval  may  be  granted  on  a 
finding  that  the  release  would  present  "no  sig- 
nificant risk  to  health  or  the  environment;”  a 
tougher  or  more  specific  standard  may  be  de- 
sirable. 

A required  study  of  the  possible  conse- 
quences of  releasing  a genetically  engineered 
organism  w'ould  be  an  important  step  in  ensur- 
ing safety.  An  impact  statement  could  be  filed 
before  permission  is  granted  to  release  the 
organism.  How'ever,  companies  and  individuals 
might  be  discouraged  from  developing  useful 
organisms  if  this  process  became  too  burden- 
some and  costly. 

F.  Congress  could  pass  legislation  regulating  all 
types  and  phases  of  genetic  engineering  from 
research  through  commercial  production. 


This  option  would  deal  comprehensively  and 
directly  with  the  risks  of  novel  molecular 
genetic  techniciues.  A s()ecific  statute  would 
eliminate  the  uncertainties  over  the  extent  to 
which  present  law  covers  particular  applica- 
tions of  genetic  engineering  and  any  concerns 
about  the  effectiveness  of  voluntary  compliance 
with  the  Guidelines.  Alternatively,  the  legisla- 
tion couki  take  the  form  of  amending  existing 
laws  to  clarify  their  applicability  to  genetic 
engineering. 

Other  molecular  genetic  technicjues,  wliile 
not  as  widely  used  and  effective  as  rDNA,  raise 
similar  concerns.  Of  the  current  techni(|ues,  cell 
fusion  is  the  prime  candidate  for  being  treated 
like  I'DNA  in  any  regulatory  framework.  No  risk 
assessment  of  this  technique  has  been  done,  and 
no  Federal  oversight  exists. 

The  principal  argument  against  this  option  is 
that  the  current  system  appears  to  be  working 
fairly  well,  and  the  limited  risks  of  the  tech- 
niques may  not  warrant  the  significantly  in- 
creased regulatory  burden  that  would  result 
from  such  legislation. 

G.  Congress  could  require  NIH  to  rescind  the 
Guidelines. 

Deregulation  w ould  have  the  adv'antage  of  al- 
lowing money  and  personnel  currently  involved 
in  implementing  the  Guidelines  at  the  Federal 
and  local  levels  to  be  used  for  other  purposes. 

There  are  several  reasons  for  retaining  the 
Guidelines.  Sufficient  scientific  concern  exists 
for  the  Guidelines  to  prohibit  certain  experi- 
ments and  to  require  containment  for  others. 
Most  experiments  can  be  done  at  the  lowest, 
least  burdensome  containment  levels.  NIH  is 
serving  an  important  role  as  a centralized  over- 
sight and  information  coordinating  body,  and 
the  system  has  been  flexible  enough  in  the  past 
to  liberalize  the  restrictions  as  evidence  in- 
dicated lower  risk  than  originally  thought. 

H.  Congress  could  consider  the  need  for  regulat- 
ing work  with  all  hazardous  micro-organisms 
and  viruses,  whether  or  not  they  are  genet- 
ically engineered. 

It  was  not  w ithin  the  scope  of  this  study  to  ex- 
amine this  issue,  but  it  is  an  emerging  one  that 
Congress  may  wish  to  consider. 


22  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Patenting  living  organisms 

On  June  16,  1980,  in  a 5-to-4  decision,  the  Su- 
preme Court  ruled  that  a human-made  micro- 
organism was  patentable  under  Federal  patent 
statutes.  The  decision  while  hailed  by  some  as 
assuring  this  country’s  technological  future  was 
at  the  same  time  denounced  by  others  as  creat- 
ing Aldous  Huxley’s  Brave  New  World.  It  will  do 
neither. 

FINDINGS 

1.  Meaning  and  Scope  of  the  Decision.— The 
decision  held  that  a patent  could  not  be  denied 
on  a genetically  engineered  micro-organism  that 
otherwise  met  the  legal  requirements  for  pat- 
entability solely  because  it  was  alive.  It  was 
based  on  the  Court’s  interpretation  of  a provi- 
sion of  the  patent  law  which  states  that  a patent 
may  be  granted  on  “.  . . any  new  and  useful  . . . 
manufacture,  or  composition  of  matter.  ...”  (35 
U.S.C.  §101) 

It  is  uncertain  whether  the  case  will  serve  as 
a legal  precedent  for  patenting  more  complex 
organisms.  Such  organisms,  however,  will  prob- 
ably not  meet  other  legal  prerequisites  to  paten- 
tability that  were  not  at  issue  here.  In  any  event, 
fears  that  the  case  would  be  legal  precedent 
sometime  in  the  distant  future  for  patenting  hu- 
man beings  are  unfounded  because  the  13th 
amendment  to  the  Constitution  absolutely  pro- 
hibits ownership  of  humans. 

2.  Impact  on  the  Biotechnology  Industry.— The 
decision  is  not  crucial  to  the  development  of  the 
industry.  It  will  stimulate  innovation  by  encour- 
aging the  dissemination  of  technical  informa- 
tion that  otherwise  would  have  been  main- 
tained as  trade  secrets  because  patents  are  pub- 
lic documents  that  fully  describe  the  inventions. 
In  addition,  the  ability  to  patent  genetically  engi- 
neered micro-organisms  will  reduce  the  risks 
and  uncertainties  facing  individual  companies 
in  the  commercial  development  of  those  orga- 
nisms and  their  products,  but  only  to  a limited 
degree  because  reasonably  effective  alterna- 
tives exist.  These  are:  1)  maintaining  the  orga- 


nisms as  trade  secrets;  2)  patenting  microbio- 
logical processes  and  their  products;  and  3)  pat- 
enting inanimate  components  of  micro-orga- 
nisms, such  as  genetically  engineered  plasmids. 

3.  Impact  on  the  Patent  Law  and  the  Patent  and 
Trademark  Office.— Because  of  the  complexity, 
reproducibility,  and  mutability  of  living  orga- 
nisms, the  decision  may  cause  some  problems 
for  a body  of  law  designed  more  for  inanimate 
objects  than  for  living  organisms.  It  raises  ques- 
tions about  the  proper  interpretation  and  appli- 
cation of  the  patent  law  requirements  of  no\  el- 
ty,  nonobviousness,  and  enablement.  In  addi- 
tion, it  raises  questions  about  how  broad  the 
scope  of  patent  coverage  on  important  micro- 
organisms should  be,  and  about  the  continuing 
need  for  two  statutes,  the  Plant  Patent  Act  of 
1930  and  the  Plant  V'arietv  Protection  Act  of 
1970.  These  uncertainties  could  result  in  in- 
creased litigation,  making  it  more  difficult  and 
costly  for  owners  of  patents  on  li\  ing  oi'ganisms 
to  enforce  their  rights. 

The  impact  on  the  Patent  and  Trademark  Of- 
fice is  not  expected  to  he  significant  in  the  luvxt 
few  years.  Although  the  number  of  patent  ap- 
plications on  micro-organisms  ha\e  almost 
doubled  during  1980,  the  approximately  200 
pending  applications  represent  less  than  0.2 
percent  of  those  processed  each  year  by  th('  Of- 
fice. While  the  number  of  such  applications  is 
expected  to  increase  in  the  next  few  yeai’s 
because  of  of  the  decision  and  de\  elopm(Mits  in 
the  field,  the  Office  should  he  ahU?  to  a(’- 
commodate  the  increase.  A few  additional  ex- 
aminers may  he  needed. 

4.  Impact  on  Academic  Research.— Because  th(' 
decision  may  encourage  academic  scientists  to 
commercialize  the  results  of  their  ix'search,  it 
may  inhibit  the  free  exchange  of  information, 
but  only  if  scientists  rely  on  track'  secrecy 
rather  than  patents  to  protect  thc'ir  iincntions 
from  competitors  in  the  marketplace*.  In  this  re- 
spect, it  is  not  clear  how  molecular  biology  dif- 
fers from  other  research  fields  w ith  commercial 
potential. 


Ch.  1 — Summary:  Issues  and  Options  • 23 


Issue  and  Options — Patenting  Living  Organisms 


ISSl'E:  I'o  what  extent  could  (]ongress 

pro\  ide  for  or  prohibit  the  pat- 
enting of  fix  ing  organisms? 

OPTIONS: 

The  SuprtMiu'  (\)uii  slated  that  it  was  under- 
taking only  the  narrow  task  of  detei'mining 
w hether  or  not  Congress,  in  enacting  the  patent 
statutes,  had  intended  a manmade  micro-orga- 
nism to  l)e  e.xcluded  from  patentahilitv  soleh' 
because  it  was  ali\e.  Moreoxer,  the  opinion 
specifically  in\  ited  Congress  to  ox  errule  the 
decision  if  it  disagreed  with  the  Cxnirt's  inter- 
pretation. Congress  can  act  to  resoh  e the  ques- 
tions left  unanswered  hy  the  Court,  oxerrule 
the  decision,  or  de\  elop  a comprehensive  statu- 
tory approach.  .Most  importantly,  Congress  can 
draw  lines;  it  can  decide  which  organisms,  if 
any,  should  he  patentable. 

A.  Congress  could  maintain  the  status  quo. 

Congress  could  choose  not  to  address  the 
issue  of  patentability  and  allow  the  law  to  he 
developed  by  the  courts.  The  adv  antage  of  this 
option  is  that  issues  will  be  addressed  as  they 
arise,  in  the  conte.xt  of  a tangible,  nonhypo- 
thetical  case. 

There  are  two  disadv  antages  to  this  option:  a 
uniform  body  of  law  may  take  time  to  develop; 
and  the  Federal  judiciary  is  not  designed  to  take 
sufficient  account  of  the  broader  political  and 
social  interests  involved. 

B.  Congress  could  pass  legislation  dealing  with 
the  specific  legal  issues  raised  by  the  Court's 
decision. 

Many  of  the  legal  questions  are  so  broad  and 
v aried  that  they  do  not  readily  lend  themselves 
to  statutory  resolution.  The  precise  meaning  of 
the  requirements  for  novelty,  nonobviousness, 
and  enablement  as  applied  to  biological  inven- 
tions will  be  most  readily  dev  eloped  on  a case- 
by-case  basis  by  the  Patent  Office  and  the 
Federal  courts.  On  the  other  hand,  some  ques- 
tions are  fairly  narrow  and  well-defined;  thus, 
they  could  be  better  resolved  by  statute.  The 
most  important  question  is  whether  there  is  a 
continuing  need  for  the  two  plant  protection 


acts  that  grant  ownership  I’ights  to  plant 
breeders  who  develop  new  and  distinct 
V arieties  of  plants. 

C.  Congress  could  mandate  a study  of  the  Plant 
Patent  Act  of  1930  and  the  Plant  Variety  Pro- 
tection Act  of  1970. 

rhese  ,'\cts  could  sei've  as  models  for  studying 
the  broader,  long-term  potential  impacts  of 
patenting  liv  ing  organisms.  Such  a study  would 
lie  timely  not  only  because  of  the  C'ourt's  deci- 
sion, hut  also  because  of  allegations  that  the 
.Acts  have  encouraged  the  planting  of  uniform 
v arieties,  loss  of  genetic  diversity,  and  increased 
concenti’ation  in  the  plant  hi'eeding  industi'y. 

D.  Congress  could  prohibit  patents  either  on  any 
living  organism  or  on  organisms  other  than 
those  already  subject  to  the  plant  protection 
Acts. 

Hy  pi’ohihiting  patents  on  any  living  or- 
ganisms, tk)ngi’ess  would  he  accepting  the 
arguments  of  those  who  consider  ownership 
rights  in  liv  ing  organisms  to  he  immoral,  or  who 
ar'e  concerned  ahoirt  other  potentially  adverse 
impacts  of  sirch  jjatents.  A total  pr'ohibition 
vvoirld  slow  hirt  not  stop  the  development  of 
molecular  genetic  techniques  and  the  biotech- 
nologv'  industr'v  becairse  there  ar^e  sever^al  good 
alternatives  for  maintaining  exclusive  contr’ol  of 
biological  inventions.  Development  would  be 
slowed  primarily  because  information  that 
might  otherwise  become  public  would  be 
withheld  as  trade  secrets.  A major  consequence 
would  be  that  desirable  products  would  take 
longer  to  reach  the  market. 

Alternatively,  Congress  could  overrule  the 
Supreme  Court’s  decision  by  amending  the  pat- 
ent law  to  prohibit  patents  on  organisms  other 
than  the  plants  covered  by  the  two  statutes 
mentioned  in  option  C.  This  would  demonstrate 
congressional  intent  that  living  organisms  could 
be  patented  only  by  specific  statute. 

E.  Congress  could  pass  a comprehensive  law  cov- 
ering any  or  all  organisms  (except  humans). 

This  option  recognizes  that  Congress  can 
draw  lines  where  it  sees  fit  in  this  area.  It  could 
specifically  limit  patenting  to  micro-organisms, 


24  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


or  it  could  (Micourage  the  breeding  of  agricul- 
turally important  animals  by  granting  patent 
i-igbts  to  brcHulers  of  new  and  distinct  breeds.  In 
the  interest  of  comprehensiveness  and  uniform- 
ity, one  statute  could  cover  plants  and  all  other 
organisms  that  Congress  desires  to  be  patent- 
able. 

Genetics  and  society 

FINDINGS 

Continued  advances  in  science  and  technol- 
ogy are  beginning  to  provide  choices  that  strain 
human  value  systems  in  areas  where  previously 
no  choice  was  possible.  Existing  ethical  and 
moral  systems  do  not  provide  clear  guidelines 
and  directions  for  those  choices.  New  programs, 
both  in  public  institutions  and  in  the  popular 
media,  have  been  established  to  explore  the 
relationships  among  science,  technology,  socie- 
ty, and  \ alue  systems,  but  more  work  needs  to 
he  done. 

Genetics— and  other  areas  of  the  biological 
sciences— have  in  common  a much  closer  rela- 
tionship to  certain  ethical  questions  than  do 
most  advances  in  the  physical  sciences  or 
engineering.  The  increasing  control  over  the 


Issues  and  Options 

Issue;  How  should  the  public  he  in- 
volved In  determining  policy  re- 
lated to  new  applications  of  ge- 
netics? 

Because  public  demands  for  involvement  are 
unlikely  to  diminish,  ways  to  accommodate 
these  demands  must  be  considered. 

OPTIONS: 

A.  Congress  could  specify  that  public  opinion 
must  be  sought  in  formulating  all  major  pol- 
icies concerning  new  applications  of  genetics, 
including  decisions  on  the  funding  of  specific 
research  projects.  A "Public  Participation 
Statement"  could  be  mandated  for  all  such 
decisions. 

B.  Congress  could  maintain  the  status  quo,  allow- 


characteristics of  organisms  and  the  potential 
for  altering  inheritance  in  a directed  fashion 
raise  again  questions  about  the  relationship  of 
humans  to  each  other  and  to  other  living  things. 
People  respond  in  different  ways  to  this  poten- 
tial; some  see  it  (like  many  predecessor  develop- 
ments in  science)  as  a challenging  opportunity, 
others  as  a threat,  and  still  others  respond  with 
vague  unease.  Although  many  people  cannot  ar- 
ticulate fully  the  basis  for  their  concern,  ethical, 
moral,  and  religious  reasons  are  often  cited. 

The  public’s  increasing  concern  about  the  ad- 
vance of  science  and  impacts  of  technology  has 
led  to  demands  for  greater  participation  in  deci- 
sions concerned  with  scientific  and  technologi- 
cal issues,  not  only  in  the  United  States  but 
throughout  the  world.  The  demands  imply  new 
challenges  to  systems  of  representative  govern- 
ment. In  every  Western  country,  new  mecha- 
nisms have  been  devised  for  increasing  citizen 
participation. 

The  public  has  already  become  in\’ol\ed  in 
decisionmaking  with  regard  to  genetics.  As  the 
science  develops,  additional  issues  in  which  the 
public  will  demand  involvement  can  he  antici- 
pated for  the  years  ahead.  The  question  then  be- 
comes one  of  how  best  to  invoke  the  public  in 
decisionmaking. 


ing  the  public  to  participate  only  when  it 
decides  to  do  so  on  its  own  initiative. 

If  option  A were  followed,  there  would  h(>  no 
cause  for  claiming  that  public  involvenu'iit  was 
inadequate  (as  occurred  after  the  first  set  of 
Guidelines  for  Recombinant  DNA  Reseai'ch  was 
promulgated).  Option  A poses  certain  [)i’ohlems: 
How  to  identify  a major  {)olicy  and  at  what  stage 
public  involvement  would  be  re(|uir(‘d.  Should 
it  take  place  only  when  technological  de\(>lop- 
ment  and  application  are  imminent,  or  at  th(> 
basic  research  stage? 

Option  B would  he  less  cumbersome  to  effect 
It  would  permit  the  estahlishiiKMit  of  ad  hoc 
mechanisms  when  necessarv. 


— Genetics  and  Society 


Ch.  1 — Summary:  Issues  and  Options  • 25 


ISSl'E:  Hou  can  I he  level  of  public 

knou  lecljj^e  concemin^J  jjenetics 
and  its  potential  be  raised? 

riiere  ai'e  some  ecliieators  wlio  beliexe  that 
too  little  time  is  sptMit  on  gtMieties  within  the 
traditional  educational  system.  Outside  the 
traditional  school  s\  stem,  a niimhei-  of  sources 
may  contrihute  to  increased  puhlic  understand- 
ing of  science  and  the  relationship  between 
science  and  societw 

Efforts  to  increase  puhlic  understanding 
should,  of  course,  he  combined  w ith  carefully 
designed  exaluation  programs  so  that  the  effec- 
ti\  eness  of  a pi'ogi'am  can  he  assessed. 

OP'nOXS: 

Proii,nims  could  h(’  dcvcloiwd  to  increase 
iniblic  underslaiuiin^  of  science  and  the  rela- 
tionship lyelween  science,  lechnoloi\\ , and 
society. 

Puhlic  und»*rstanding  ol  science  in  today's 
world  is  ('ssenlial.  and  th('re  is  concern  about 
th('  ad<*{|uacy  of  the  public's  know  U‘dg('. 

B.  Programs  could  he  established  to  monitor  the 
level  of  public  understanding  of  genetics  and 
of  science  in  general,  and  to  determine  wheth- 
er public  concern  with  decisionmaking  in 
science  and  technology'  is  increasing. 

Selecting  this  option  would  indicate  that 
there  is  need  for  additional  information,  and 
that  Congress  is  interested  in  invoking  the 
public  index  eloping  science  policy. 

C.  The  copyright  laws  could  be  amended  to  per- 
mit schools  to  videotape  television  programs 
for  educational  purposes. 

Under  current  copyright  law,  x ideotaping  tel- 
evision programs  as  they  are  being  broadcast 
may  infringe  on  the  rights  of  the  program’s 
owner,  generally  its  producer.  The  legal  status 
of  such  tapes  is  presently  the  subject  of  litiga- 
tion. 

In  favor  of  this  option,  it  should  be  noted  that 
many  of  the  programs  are  made  at  least  in  part 
with  public  funds.  Removing  the  copyright  con- 
straint on  schools  would  make  these  programs 
more  available  for  another  public  good,  educa- 


tion. On  the  other  hand,  this  option  could  have 
significant  economic  conseciuences  to  the  copy- 
right owner,  whose  market  is  often  limited  to 
educational  institutions. 

ISSUE:  Sboiild  Congress  begin  prepar- 

ing nou'  to  resolve  issues  tbat 
bave  not  yet  aroused  mucb  pub- 
lic debate  but  wbicb  may  in  tbe 
future? 

.As  scientific  understanding  of  genetics  and 
the  ability  to  manipulate  inherited  character- 
istics develo[)s,  society  may  face  some  difficult 
c|uestions  that  could  involve  tradeoffs  between 
individual  freedom  and  the  needs  of  society. 
I'his  will  he  increasingly  the  case  as  genetic 
technologies  are  a|)plied  to  humans.  Develop- 
ments are  occurring  rapidly.  Recombinant  DNA 
technologx’  was  develoj)ed  in  the  1970’s.  In  the 
spring  of  1980,  investigators  succeeded  in  the 
first  gene  replacement  in  mammals;  in  the  fall 
of  1980,  the  first  gene  substitution  in  humans 
was  attempted. 

Although  this  study  was  restricted  to  nonhu- 
man applications,  many  people  assume  from 
these  and  other  examples  that  what  can  be  done 
with  lower  animals  can  be  done  with  humans 
and  will  he.  Therefore,  some  action  might  be 
taken  to  better  prepare  society  for  decisions  on 
the  application  of  genetic  technologies  to 
humans. 

OPTIONS: 

A.  A commission  could  be  established  to  identify 
central  issues,  the  probable  time  frame  for  ap- 
plication of  various  genetic  technologies  to 
humans,  and  the  probable  effects  on  society, 
and  to  suggest  courses  of  action.  The  commis- 
sion might  also  consider  the  related  area  of 
how  participatory  democracy  might  be  com- 
bined with  representative  democracy  in  deci- 
sionmaking. 

B.  The  life  of  the  President's  Commission  could 
be  extended  for  the  study  of  Ethical  Problems 
in  Medicine  and  Biomedical  and  Behavioral  Re- 
search, for  the  purpose  of  addressing  these 
issues. 


26  • Impacts  of  Afiplied  Genetics — Micro-Organisms,  Plants,  and  Animals 


I his  1 1 -member  Commission  was  established 
in  Novembei'  1978  and  terminates  on  December 
91,  1982.  It  could  be  asked  to  broaden  its  cover- 
age to  additional  areas.  This  would  require  that 
the  life  span  of  the  commission  be  extended  and 
additional  funds  be  appropriated. 

A potential  disadvantage  to  using  the  existing 
commission  to  address  societal  issues  associated 
with  genetic  engineering  is  that  a number  of 


issues  already  exist,  and  more  are  likely  to  arise 
in  tbe  years  ahead.  Yet  there  are  also  other 
issues  in  medicine  and  biomedical  and  beba\  - 
ioral  research  not  associated  with  genetic  engi- 
neering that  also  need  review.  Whether  all 
these  issues  can  be  addressed  by  one  commis- 
sion should  be  considered.  Comments  from  tbe 
existing  commission  would  assist  in  deciding  tbe 
most  appropriate  course  of  action. 


o 


Chapter  2, 

Introduction 


Chapter  2 


Page 


The  Origins  of  Genetics 29 

Genetics  in  the  20th  Century 33 

The  Riddle  of  the  Gene 33 

The  Genetic  Code 37 

Developing  Genetic  Technologies 39 

The  Basic  Issues ' 43 

How  Will  Applied  Genetics  Be  Used? 43 

What  Are  the  Dangers? 43 


Figures 


Figure  No.  Page 

5.  The  Inheritance  Pattern  of  Pea  Color 30 

6.  Chromosomes 32 


7.  The  Griffith  Experiment 34 

8.  The  Structure  of  DNA 36 

9.  Replication  of  DNA 37 

10.  The  Genetic  Code 38 

11.  The  Expression  of  Genetic  Information  in 

the  Cell 39 

12.  Transduction;  The  Transfer  of  Genetic 
Material  in  Bacteria  by  Means  of  Viruses  ....  39 

13.  Conjugation:  The  Transfer  of  Genetic 

Material  in  Bacteria  by  Mating 40 

14.  Recombinant  DNA:  The  Technique  of 

Recombining  Genes  From  One  Species  With 
Those  From  Another 41 

15.  An  Example  of  How  the  Recombinant  DNA 
Technique  May  Be  Used  To  Insert  New 

Genes  Into  Bacterial  Cells 42 


Chapter  2 

Introduction 


Humankiiul  is  gaining  an  increasing  under- 
standing ot  heredity  and  \ ariation  among  Ii\  ing 
tilings— the  science  of  genetics.  I his  report  e.\- 
amines  hotli  the  critical  issues  arising  from  the 
science  and  technologies  that  spring  trom  ge- 
netics, and  the  potential  impacts  of  these  ad- 
vances on  society.  Ihey  ai'e  the  most  rapidly 
progressing  areas  of  human  know  ledge  in  the 
world  today. 

(lenetic  technologies  e.xist  onl\  within  the 
largei'  conte.xt  of  a maturing  science.  The  key  to 
planning  for  their  potential  is  understanding 
not  simjih  a [larticulai’  technologv',  oi'  breeding 
[iiogram,  or  new  opportunity  foi'  investment, 
hut  how  the  field  of  genetics  works  and  how  it 
intei'acts  with  society  as  a vv  hole. 

The  technologies  that  this  I'eport  assesses  can 
he  expected  to  hav  e pervasiv  e effects  on  life  in 
the  future.  They  touch  on  the  most  fundamen- 
tal and  intimate  needs  of  mankind:  health  care, 
supplies  of  food  and  enei'gv , and  reproduction. 
.\t  the  same  time,  they  trigger  concerns  in  areas 


The  origins  of  genetics 

For  the  past  10,000  years,  a period  encom- 
passing less  than  one-half  of  1 percent  of  man’s 
time  on  Earth,  the  human  race  has  developed 
under  the  impetus  of  applied  genetics.  As  tech- 
niques for  planning,  cultivating,  and  storing 
crops  replaced  subsistence  hunting  and  forag- 
ing, the  character  of  humanity  changed  as  well. 
From  the  domestication  of  animals  to  the  devel- 
opment of  permanent  settlements,  from  the  rise 
of  modern  science  to  the  dawn  of  biotech- 
nology, the  genetic  changes  that  mankind  has 
directed  have,  in  turn,  affected  the  nature  of  his 
society. 

Applied  genetics  depends  on  a fundamental 
principle— that  organisms  both  resemble  and 
differ  from  their  parents.  It  must  have  required 
great  faith  on  the  part  of  Neolithic  man  to  bury 


etiually  as  important:  the  dwindling  su|)|ilies  of 
natural  resources,  the  risks  involved  in  basic 
and  applied  scientific  research  and  develop- 
ment, and  the  nature  of  innovation  itself. 

•As  always,  some  decisions  concerning  the  use 
of  the  new  technologies  will  he  made  by  the 
marketplace,  while  others  will  he  made  by  var- 
ious institutions,  both  public  and  pi’ivate.  In  the 
coming  years,  the  public  and  its  rei)resentatives 
in  (Congress  and  other  gov  ernmental  bodies  will 
be  called  on  to  make  difficult  decisions  because 
of  society’s  knowledge  about  genetics  and  its 
capabilities. 

Fhis  report  does  not  make  recommendations 
noi'  does  it  attempt  to  resolve  conflicts.  Kather, 
it  clarifies  the  bases  for  making  judgments  by 
defining  the  likely  impacts  of  a group  of  technol- 
ogies and  tracing  their  economic,  societal,  legal, 
and  ethical  implications.  The  new  genetics  will 
be  influential  for  a long  time  to  come.  Although 
it  will  continue  to  change,  it  is  not  too  early  to 
begin  to  monitor  its  course. 


perfectly  good  grain  during  one  season  in  the 
hope  of  growing  a new  crop  several  months 
later— faith  not  only  that  the  seed  would  indeed 
return,  but  that  it  w ould  do  so  in  the  form  of  the 
same  grain-producing  crop  from  which  it  had 
sprung.  This  permanence  of  form  from  one 
generation  to  the  next  has  been  scientifically 
understood  only  within  the  past  century,  but 
the  understanding  has  transformed  vague  be- 
liefs in  the  inheritance  of  traits  into  the  science 
of  genetics,  and  rule-of-thumb  animal  and  plant 
breeding  into  the  modern  manipulations  of 
genetic  engineering. 

The  major  conceptual  boost  for  the  science 
of  genetics  required  a shift  in  perspective, 
from  the  simple  observation  that  characteristics 


29 


30  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


passed  from  parents  to  offspring,  to  a study  of 
the  underlying  agent  by  which  this  transmission 
is  accomplished.  That  shift  began  in  the  garden 
of  Gregor  Mendel,  an  obscure  monk  in  mid-19th 
century  Austria.  By  analyzing  generations  of 
controlled  crosses  between  sweet  pea  plants, 
Mendel  was  able  to  identify  the  rudimentary 
characteristics  of  what  was  later  termed  the 
gene. 

Mendel  reasoned  that  genes  were  the  vehicle 
and  repository  of  the  hereditary  mechanism, 
and  that  each  inherited  trait  or  function  of  an 
organism  had  a specific  gene  directing  its  devel- 
opment and  appearance.  An  organism’s  observ- 
able characteristics,  functions,  and  measurable 
properties  taken  together  had  to  be  based  some- 
how on  the  total  assemblage  of  its  genes. 

Mendel’s  analysis  showed  that  the  genes  of 
his  pea  plants  remained  constant  from  one  gen- 
eration to  the  next,  but  more  importantly,  he 
found  that  genes  and  observable  traits  were  not 
simply  matched  one-for-one.  There  were,  in 
fact,  two  genes  involved  in  each  trait,  with  a 
single  gene  contributed  by  each  parent.  When 
the  genes  controlling  a particular  trait  are  iden- 
tical, the  organism  is  homozygous  for  that  trait; 
if  they  are  not,  it  is  heterozygous. 

In  the  Mendelian  crosses,  homozygous  plants 
always  retained  the  expected  characteristics. 
But  heterozygous  plants  did  not  simply  display  a 
mixture  of  their  different  genes;  one  of  the  two 
tended  to  predominate.  Thus,  when  homozy- 
gous yellow-seed  peas  were  crossed  with  homo- 
zygous green-seed  plants,  all  the  offspring  were 
now  heterozygous  for  seed  color,  possessing  a 
“green”  gene  from  one  parent  and  a "yellow” 
from  the  other.  Yet  all  of  them  turned  out  to  be 
indistinguishable  from  the  yellow-seed  parent: 
Yellow-seed  color  in  peas  was  dominant  to 
green. 

But  even  though  the  offspring  resembled 
their  dominant  parent,  they  could  be  shown  to 
contain  a genetic  difference.  For  when  the  het- 
erozygotes were  now  crossed  with  each  other,  a 
certain  number  of  recessive  green-seed  plant 
again  appeared  among  the  offspring.  This  oc- 
curred whenever  an  offspring  was  endowed 
with  a pair  of  genes  that  was  homozygous  for 


the  green-seed  trait— and  it  occurred  at  a rate 
consistent  with  the  random  selection  of  one  of 
two  genes  from  each  parent  for  passage  to  the 
new  generation.  (See  figure  5.) 

Genes  were  real— Mendel’s  work  made  that 
clear.  But  where  were  they  located,  and  what 
were  they?  The  answer,  lay  within  the  nucleus 
of  the  cell.  Unfortunately,  most  of  the  contents 
of  the  nucleus  were  unobtainable  by  biologists 
in  Mendel’s  time,  so  his  published  findings  were 
ignored.  Only  during  the  last  decades  of  the 
19th  century  did  improved  microscopes  and 
new  dyes  permit  cells  to  be  observed  with  an 
acuity  never  before  possible.  And  only  by  the 


Figure  5.— The  Inheritance  Pattern  of  Pea  Color 

Y = yellow  gene  g = green  gene 

Homozygous  yellow-seed  peas  have  the  genetic  compost- 
tion;  YY. 

|N^WS|fgous  green-seed  peas  have  the  genetic  carspoBt' 
ion:  gg. 

Each  parent  contributes  only  one  seed-color  gene  to  the  off- 
spring. When  the  two  YY  and  gg  homozygotes  are  crossed, 
the  genetic  composition  of  all  offspring  is  Yg: 


All  Yg  offspring  are  heterozygous,  and  all  have  yellow 
seeds,  indicating  that  the  Y yellow  gene  is  dominant  over 
the  g green  gene. 

When  these  Yg  heterozygotes  are  crossed  with  each  other: 


Vi  of  the  total  are  homozygous  YY,  having  yejlow  seeds 
V4  ofjhe  total  are  homozygous  gg,'  having  srmh  aa«^  | 
Vi  of  the  total  are  heterozygous  Yg,  having  yellow  seeds 

Thus,  % of  these  offspring  will  have  yellow  seeds,  but  their 
individual  genetic  composition,  YY  of  Yg,  may  be  different 

SOURCE:  Office  of  Technology  Assessment. 


Ch.  2 — Introduction  • 31 


beginning  ot  the  20tli  centiirv  did  scientists 
rediscover  Mendel’s  work  and  begin  to  ap[)re- 
ciate  fully  the  significance  of  the  cell  nucleus 
and  its  contents. 

K\en  in  the  earliest  microscopic  studies, 
boue\er,  certain  cellular  com[)onents  stood 
out;  they  were  deeply  stained  by  added  dye.  As 
a result,  they  were  dubbed  “coloretl  bodies,”  oi' 
chromosomes.  Chromosomes  v\ere  seen  rela- 
ti\ely  rarely  in  cells,  with  most  cells  showing 
just  a central  tlark  nucleus  surrounded  by  an 
e.xtensive  light  grainy  cytoplasm.  But  periodi- 
cally the  nucleus  seemed  to  disappear,  leaving 
in  its  place  long  thready  material  that  con- 
solidated to  form  the  chromosomal  bodies.  (See 
figure  6a.)  Once  formed,  the  chromosomes 
assembled  along  the  middle  of  the  cell,  copied 
themsek  es,  and  then  mo\  ed  apart  w bile  the  cell 
pinched  itself  in  half,  trapping  one  set  of 
chromosomes  in  each  of  the  two  hakes.  I hen 
the  chromosomes  themsekes  seemed  to  dis- 
soke  as  two  new  nuclei  appeared,  one  in  each 
of  the  tw  o newly  formed  cells.  (See  figure  6h.) 

Thus,  the  same  number  of  chromosomes  ap- 
peared in  precisely  the  same  form  in  e\ery  cell 
of  an  organism  e.xcept  the  germ,  or  sex,  cells. 
Furthermore,  the  chromosomes  not  only  re- 
mained constant  in  form  and  number  from  one 
generation  to  the  next,  hut  were  inherited  in 
pairs.  They  were,  in  short,  manifesting  all  the 


traits  that  Mendel  had  prescribed  for  genes 
almost  three  decades  earlier.  By  the  beginning 
of  the  20th  century,  it  was  clear  that  chromo- 
somes w'ere  of  central  importance  to  the  life  his- 
tory of  the  cell,  acting  in  some  unspecified  man- 
ner as  the  vehicle  for  the  Mendelian  gene. 

If  this  conclusion  was  strongly  implied  by  the 
e\  ents  of  cell  di\  ision,  it  became  obvious  when 
I'eproduction  in  whole  organisms  was  analyzed. 
It  had  been  established  by  the  latter  part  of  the 
19th  century  that  the  germ  cells  of  plants  and 
animals— |)ollen  and  o\  um,  sperm  and  egg— ac- 
tually fuse  in  the  [process  of  fertilizaton.  Germ 
cells  differ  fi’om  other  body  cells  in  one  impor- 
tant resj)ect— they  contain  only  half  the  usual 
number  of  chromosomes.  This  chromosome 
baking  within  the  cell  was  apparently  done 
\'ery  precisely,  for  e\'ery  sperm  and  egg  con- 
tained exactly  one  representative  from  each 
chromosome  pair.  When  the  two  germ  cells 
then  fused  during  fertilization,  the  offspring 
were  supplied  with  a fully  I’econstituted  chro- 
mosome complement,  half  from  each  parent. 
C^learly,  chromosomes  were  the  material  link 
from  one  generation  to  the  next.  Somewhere 
locked  within  them  was  the  substance  of  both 
heredity— the  fidelity  of  traits  between  genera- 
tions; and  diversity— the  potential  for  genetic 
\ ariation  and  change. 


32  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Figure  6.— Chromosomes 


<*«• 


: HI'S* 


J 


Photo  credit:  Professor  Judith  Lengyei,  Molecular  Biology  Institute,  UCLA 


Optical  micrograph  of  chromosomal  material  from  the  salivary  gland  of  the  larva  of  the 
common  fruit  fly,  Drosophila  rnelanogaster 


6b.  In  Step  1,  the  chromosome  bodies  are  still  uncondensed. 

In  Steps  2 and  3,  the  chromosomes  condense  into  thread-like  bodies  and  align  themselves  near  the  center  of  the  cell. 

In  Steps  4 and  5,  the  chromosomes  begin  to  separate  and  are  pulled  to  the  opposite  poles  of  the  cell. 

In  Step  6,  the  chromosomes  return  to  an  uncondensed  state  and  the  cell  begins  to  constrict  about  the  middle  to  form 
two  new  cells. 


SOURCE:  Office  of  Technology  Assessment. 


Ch.  2 — Introduction  • 33 


Genetics  in  the  20th  century 


During  the  first  few  decades  of  the  2()tli  cen- 
tury. scientists  seardied  for  progressi\ely 
simpler  experimental  organisms  to  clai'ity  pro- 
gressi\  ely  more  complex  genetic  concepts.  First 
was  Thomas  Hunt  .Morgan’s  Drosop/j;7a— gnat- 
sized fruit  flies  v\  ith  hulhous  eyes.  These  insects 
ha\e  a simple  array  of  four  easily  distinguish- 
able chromosome  paii's  per  cell.  They  repro- 
duce rapidly  and  in  large  numbers  under  the 
simplest  of  laboratory  conditions,  supplying  a 
new  generation  e\ery  month  or  so.  Thus,  re- 
searchers could  carry  out  an  enormous  number 
of  crosses  employing  a whole  catalog  of  dif- 
ferent fruit  tlv  traits  in  a relativ  ely  brief  time. 

It  became  ohxious  from  the  extensi\e  Dros- 
ophila data  that  certain  traits  were  more  likely 
to  be  inherited  together  than  others.  \'ellow 
bodies  and  ruby  eyes,  for  instance,  almost  al- 
ways went  together,  w ith  both  in  turn,  appear- 
ing more  frequently  than  expected  with  the 
trait  known  as  "forked  bristles. " .All  three  traits, 
however,  showed  up  onlv  randomly  with 
curved  wings.  Certain  genes  thus  seemed  to  be 
linked  to  one  another.  The  entire  Drosophila 
genome,  in  fact,  fell  into  four  distinct  linkage 
groups.  The  physical  basis  for  these  groups,  not 
surprisingly,  consisted  of  the  four  fruit  fly 
chromosomes.  Linked  genes  behaved  as  they 
did  because  they  were  located  on  the  same 
chromosome. 

Soon,  scientists  learned  that  they  could  not 
only  assign  particular  genes  to  particular  Droso- 
phila chromosomes  but  could  identify  tbe  rela- 
tive  locations  of  different  genes  on  a given 
chromosome.  This  gene  mapping  was  possible 


The  riddle  of  the  gene 

W ith  all  this  research,  nobody  yet  knew  what 
the  gene  was  made  of.  The  first  evidence  that 
it  consisted  of  deoxyribonucleic  acid  (DNA) 
emerged  from  the  work  of  Oswald  Avery,  Colin 
MacLeod,  and  Maclyn  McCarty  at  the  Rockefel- 
ler Institute  in  New  York  in  the  early  1940’s. 
Avery’s  group  took  as  its  starting  point  some  in- 


hecause linkage  itself  was  not  permanent, 
linked  genes  sometimes  separated.  For  instance, 
w hile  yellow  bodies,  ruby  eyes,  and  forked  bris- 
tles were  all  linked  traits,  tbe  first  two  stayed 
together  far  more  frequently  than  either  did 
with  the  third. 

The  degree  of  linkage  between  two  genes  was 
hypothesized  to  be  directly  proportional  to  the 
distance  between  them  on  the  chromosome, 
mainly  because  of  a unic|ue  event  that  occurs 
during  the  development  of  germ  cells.  Before 
the  normal  chromo.some  number  is  halved,  the 
chromosomes  crowd  together  in  the  center  of 
the  cell,  coiling  tightly  around  each  other,  prac- 
tically fusing  along  their  entire  length.  It  is  in 
this  state  that  crossing-over  (or  natural  recombi- 
nation)—the  actual  physical  exchange  of  parts 
between  chromosomes— occurs.  No  chromo- 
some emerges  from  the  exchange  in  the  same 
condition  as  before;  the  lengths  of  chromo- 
somes are  reshuffled  before  being  transferred 
to  the  next  generation. 

The  idea  of  linkage  meant  that  Mendel’s  for- 
mulations had  to  be  modified.  Clearly,  genes 
were  not  completely  independent  units.  Further 
work  with  Drosophila  in  the  1920’s  showed  that 
genes  were  also  not"  permanent  and  could 
change  over  time.  Although  natural  mutations 
occurred  at  a very  slow  rate,  exposing  fruit  flies 
to  X-rays  accelerated  their  frequency  enor- 
mously. Exposure  of  a parental  fly  population 
led  to  an  array  of  new  traits  among  their  off- 
spring-traits which,  if  they  w'ere  neither  lethal 
nor  sterilizing,  could  be  passed  from  one  gen- 
eration to  the  next. 


triguing  observations  made  a decade  earlier  by 
a British  physician,  Fred  Griffith.  He  had 
worked  wdth  two  types  of  pneumococcus  (the 
bacteria  responsible  for  pneumonia)  and  with 
two  different  bacteria  within  each  type.  One 
bacterium  in  each  type  was  coated  in  a polysac- 
charide capsule;  the  other  was  bare.  Bare  bac- 


34  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


teria  gave  rise  only  to  bare  progeny,  while  those 
with  capsules  produced  only  encapsulated 
forms.  Only  the  encapsulated  forms  of  both 
types  II  and  III  could  cause  disease;  bare  bac- 
teria were  benign.  (See  figure  7a.)  But  when 
Griffith  took  some  encapsulated  type  III  bacteria 
that  had  been  killed  and  rendered  harmless  and 
mixed  them  with  bare  bacteria  of  type  II,  the 
presumably  safe  mixture  became  virulent:  Mice 
injected  with  it  died  of  a massive  pneumonia  in- 
fection. Bacteria  recovered  from  these  animals 
were  found  to  be  of  type  II— the  only  living  bac- 
teria the  mice  had  received— now  wrapped  in 
type  III  capsules.  (See  figure  7b.) 

Avery’s  group  recognized  Griffith’s  finding  as 
a genetic  phenomenon;  the  dead  type  III  bacte- 
ria must  have  delivered  the  gene  for  making 
capsules  into  the  genetic  complement  of  the 
living  type  II  recipients.  By  meticulous  research, 
Avery’s  group  found  that  the  substance  which 
caused  the  genetic  transformation  was  DNA. 

It  had  been  in  1868,  just  3 years  after  Mendel 
had  published  his  findings,  that  DNA  was  dis- 
covered by  Friedrich  Miescher.  It  is  an  extreme- 
ly simple  molecule  composed  of  a small  sugar 
molecule,  a phosphate  group  (a  phosphorous 
atom  surrounded  by  four  oxygen  atoms),  and 
four  kinds  of  simple  organic  chemicals  known 
as  nitrogenous  (nitrogen-containing)  bases.  To- 
gether, one  sugar,  one  phosphate,  and  one  base 
form  a nucleotide— the  basic  structural  unit  of 
the  large  DNA  molecule.  Because  it  is  so  simple, 
DNA  had  appeared  to  be  little  more  than  a 
monotonous  conglomeration  of  simple  nucleo- 
tides to  scientists  in  the  early  20th  century.  It 
seemed  unlikely  that  such  a prosaic  molecule 
could  direct  the  appearance  of  genetic  traits 
while  faithfully  reproducing  itself  so  that  in- 
formation could  be  transferred  between  gen- 
erations. Although  Avery’s  results  seemed  clear 
enough,  many  were  reluctant  to  accept  them. 

Those  doubts  were  finally  laid  to  rest  in  a 
brief  report  published  in  1953  by  James  Watson 
and  Francis  Crick.  By  using  X-ray  crystallo- 
graphic techniques  and  building  complex  mod- 
els—and  without  ever  having  actually  seen  the 
molecule  itself— Watson  and  Crick  reported  that 
they  had  discovered  a consistent  scientifically 
sound  structure  for  DNA. 


Figure  7.— The  Griffith  Experiment 

7a.  There  are  two  types  of  pneumococcus,  each  of  which 
can  exist  in  two  forms: 

Type  II  Type  III 

y\ 

®ii  *^iii  ®ni 

where  R represents  the  rough,  nonencapsulated,  benign 
form; and 

S represents  the  smooth,  encapsulated,  virulent 
form. 

7b.  The  experiment  consists  of  four  steps: 


Virulent  strain  (1) 

Mice  injected  with  the  virulent  Sm  die. 


Living 


Nonvirulent 

strain 


(2) 


Mice  injected  with  nonvirulent  Rn  do  not  become  infected. 


S|ii 

Virulent 

strain, 

heat-killed 


Living 


The  virulent  Sm  is  heat-killed.  Mice  injected  with  it  do  not 
die. 


When  mice  are  injected  with  the  nonvirulent  R,  and  thp 
heat-killed  Sm,  they  die.  Type  II  bacteria  wrapped  m type  III 
capsules  are  recovered  from  these  mice 

SOURCE:  Office  of  Technology  Assessment 


Ch.  2 — Introduction  • 35 


I'he  structure  that  Cirick  and  V\'atson  uncov- 
ered sohed  part  of  the  genetic  puzzle.  Accord- 
ing to  them,  the  phosphates  and  sugars  formed 
two  long  chains,  or  backbones,  with  one  nitrog- 
enous base  attached  to  each  sugar.  The  two 
backbones  were  held  together  like  the  supports 
of  a ladder  by  weak  attractions  between  tbe 
bases  protruding  from  the  sugar  molecules.  Of 
the  four  different  nitrogenous  bases— adenine, 
thymine,  guanine,  and  cytosine— attractions  e.\- 
isted  only  between  adenine(.A)  aiid  thymine(T), 
and  between  guanineKi)  and  cytosine(C').  (See 
figure  8a)  Thus,  if  a stretch  of  nucleotides  on 
one  backbone  ran: 

.\-T-(.-c:-T-r-.\ -.\ 

the  other  backbone  had  to  contain  the  directly 
opposite  complementary  setjuence: 

T-.-\-C  (;-.\  A-  r-  r. . 

The  complementary  pairing  between  bases  run- 
ning down  the  center  of  the  long  molecule  was 
responsible  for  holding  together  the  two  other- 
wise independent  chains.  (See  figure  8b.)  Thus, 
the  Di\A  molecule  was  rather  like  a zipper,  with 
the  bases  as  the  teeth  and  the  sugar-phosphate 
chains  as  the  strands  of  cloth  to  which  each  zip- 
per half  was  sewn.  Crick  and  \\  atson  also  found 
that  in  the  presence  of  water,  the  two  poly- 
nucleotide chains  did  not  stretch  out  to  full 
length,  but  twisted  around  each  other,  forming 
what  has  undoubtedly  become  the  most  glori- 
fied structure  in  the  history  of  biology— the  dou- 
ble helix.  (See  figure  8c.) 

The  structure  was  scientifically  elegant.  But  it 
was  received  enthusiastically  also  because  it  im- 
plied how  DNA  worked.  As  Crick  and  Watson 
themselves  noted: 


If  the  actual  order  of  the  bases  on  one  of  the 
pair  of  chains  were  given,  one  could  write  down 
the  exact  order  of  the  bases  on  the  other  one, 
because  of  the  specific  pairing.  Thus  one  cliain 
is,  as  it  were,  the  complement  of  the  other,  and 
it  is  this  feature  which  suggests  how  the  desoxy- 
ribonucleic acid  molecule  might  duplicate 
itself.' 

V\'hen  a double-stranded  DNA  molecule  is  un- 
zipped, it  consists  of  two  separate  nucleotide 
chains,  each  with  a long  stretch  of  unpaired 
bases.  In  the  presence  of  a mixture  of  nucleo- 
tides, each  base  attracts  its  complementary 
match  in  accordance  with  the  inherent  affinities 
of  adenine  for  thymine,  thymine  for  adenine, 
guanine  for  cytosine,  and  cytosine  for  guanine. 
The  result  of  this  re[)lication  is  two  DNA  mole- 
cules, both  precisely  identical  to  each  other  and 
to  the  original  molecule— which  explains  the 
faithful  duplication  of  the  gene  for  passage  from 
one  generation  to  the  next.  (See  figure  9.) 

Crick  and  Watson’s  work  solved  a major  rid- 
dle in  genetic  research.  Because  George  Beadle 
and  Edward  Tatum  had  recently  discovered 
that  genes  control  the  appearance  of  specific 
proteins,  and  that  one  gene  is  responsible  for 
producing  one  specific  protein,  scientists  now 
knew  what  the  genetic  material  was,  how  it  rep- 
licated, and  what  it  produced.  But  they  had  yet 
to  determine  how  genes  expressed  themselves 
and  produced  proteins. 


'James  D.  Watson  and  Francis  Crick,  "Genetical  Implications  of 
the  Structures  of  Deoxyribose  Nucleic  Acid,"  Nature  171,  1953.  pp. 
737-8. 


36  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


Figure  8.— The  Structure  of  DNA 


8b.  The  four  bases  form  the  four  letters  in  the  alphabet  of 
the  genetic  code.  The  sequence  of  the  bases  along  the 
sugar-phosphate  backbone  encodes  the  genetic  in- 
formation. 


A schematic  diagram  of  the  DNA  double  helix.  A three-dimensional  representation  of  the  DNA  double  helix 

8c.  The  DNA  molecule  is  a double  helix  composed  of  two  chains.  The  sugar-phosphate  backbones  twist  around  the  out 
side,  with  the  paired  bases  on  the  inside  serving  to  hold  the  chains  together. 

SOURCE;  Office  of  Technology  Assessnnent. 


c/7.  2 — Introduction  • 37 


Figure  9.— Replication  of  DNA 

Old  Old 


When  DNA  replicates,  the  original  strands  unwind  and 
serve  as  templates  for  the  building  of  new  complementary 
strands.  The  daughter  molecules  are  exact  copies  of  the 
parent,  with  each  having  one  of  the  parent  strands. 


SOURCE:  Office  of  Technology  Assessment. 


The  genetic  code  

Proteins  are  the  basic  materials  of  cells.  Some 
proteins  are  enzymes,  which  catalyze  reactions 
within  a cell.  In  general,  for  every  chemical  re- 
action in  a lix’ing  organism,  a specific  enzyme  is 
required  to  trigger  the  process.  Other  proteins 
are  structural,  comprising  most  of  the  raw  ma- 
terial that  forms  cells. 


Ironically,  proteins  are  far  more  complex  and 
diverse  than  the  four  nucleotides  that  help 
create  them.  Proteins,  too,  are  long  chains  made 
up  of  small  units  strung  together.  In  this  case, 
however,  the  units  are  amino  acids  rather  than 
nucleotides— and  there  are  20  different  kinds  of 
amino  acids.  Since  an  average  protein  is  a few 


38  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


hundred  amino  acids  in  length,  and  since  any 
one  ot  20  amino  acids  can  fill  each  slot,  the  num- 
her  of  possible  proteins  is  enormous.  Neverthe- 
less, each  protein  requires  the  strictest  ordering 
of  amino  acids  in  its  structure.  Changing  a 
single  amino  acid  in  the  entire  sequence  can 
drastically  change  the  protein's  character. 

It  was  now  possible  for  scientists  to  move 
nearer  to  an  appreciation  of  how  genes  func- 
tioned. First  had  come  the  recognition  that  DNA 
determined  protein;  now  it  was  evident  that  the 
sec|uence  of  nucleotides  in  DNA  determined  a 
linear  sequence  of  amino  acids  in  proteins. 

By  the  early  1980’s,  the  way  proteins  were 


manufactured,  how  their  synthesis  was  regu- 
lated, and  the  role  of  DNA  in  both  processes 
were  understood  in  considerable  detail.  The 
process  of  transcribing  DNA’s  message— carry- 
ing the  message  to  the  cell’s  miniature  pi'otein 
factories  and  building  proteins— took  place 
through  a complex  set  of  reactions.  Kach  amino 
acid  in  the  protein  chain  was  represented  by 
three  nucleotides  from  the  DNA.  That  thi'ee- 
hase  unit  acted  as  a word  in  a DNA  sentc'nce 
that  spelled  out  each  |)rotein— the  genetic  codcv 
(See  figure  10.) 

Thiough  the  genetic  ('ode,  an  entire*  gene— a 
linear  assemblage  of  nuclen)tides— could  now  he 


Figure  10.— The  Genetic  Code 


SECOND 

BASE 


THIRD  BASE 


ser 


pro 


thr 


ala 


SECOND 

BASE 

T 


THIRD  BASE 


A 


E 


his 


asn 


asp 


tyr 


his 


asn 


asp 


och’ 


gin 


lys 


glu 


I 1 

amb* 

I I 


gin 


lys 


glu 


SECOND 

BASE 


G 


cys 


arg 


ser 


giy 


ser 


pro 


thr 


ala 


ser 


pro 


thr 


ala 


ser 


pro 


thr 


ala 


THIRD  BASE 


cys 


arg 


ser 


giy 


end* 


arg 


arg 


giy 


trp 


arg 


arg 


giy 


*och  (ochre);  amb  (amber),  and  end  are  stop  signal  for  translation,  i.e., 
signal  the  end  of  synthesis  of  the  protein  chain. 


Amino  acid 

Three-letter 

symbol 

alanine 

ala 

arginine 

arg 

asparagine 

asn 

aspartic  acid 

asp 

asn  and/or  asp 

asx 

cysteine 

cys 

glutamine 

gin 

glutamic  acid 

glu 

gin  and/orglu 

glx 

glycine 

giy 

histidine 

his 

isoleucine 

ileu 

leucine 

leu 

lysine 

lys 

methionine 

met 

phenylalanine 

phe 

proline 

pro 

serine 

ser 

threonine 

thr 

tryptophan 

trp 

tyrosine 

tyr 

valine 

val 

Each  amino  acid  is  determined  by  a 
three  letter  code  (A,  G,  T,  or  C)  along 
the  DNA.  If  the  first  letter  in  the  code 
is  A,  the  second  is  T.  and  third  is  A, 
the  amino  acid  will  be  tyrosine  (or  tyr) 
in  the  complete  protein  molecule.  For 
leucine  (or  leu),  the  code  is  GAT,  and 
so  forth.  The  dictionary  above  gives 
the  entire  code. 


ATA  GAT  AGA  TAG  ATAG 

V V V V V 

tyr  - leu  • ser  • ileu  • tyr 


SOURCE:  Office  of  Technology  Assessment. 


Ch.  2— Introduction  • 39 


read  like  a hook.  By  tlie  ld7()'s,  l•(>seal'ehers  liad 
leai'ned  to  read  the  code  of  certain  |)rot(*ins, 
sMithesize  tlieir  1).\  \,  and  insei't  the  l)\  \ into 
hactei'ia  so  that  the  protein  couUi  he*  pioduced. 
(See  figure  11.) 

Meanwhile,  othei’  scientists  were  studying 
the  genetics  ot  xii'iises  and  hactei'ia.  The  com- 
hination  ot  these  studi(\s  with  those  iinestigat- 
ing  the  gent'tic  code  led  to  the  inno\ations  ol 
genetic  engineei'ing. 


Figure  11.— The  Expression  of  Genetic  Information 
in  the  Cell 

DNA  m mRNA  ■ » Protein 

(Transcription)  (Translation) 

process  process 

The  "central  dogma”  of  molecular  biology:  DNA  in  the 
genes  is  transcribed  into  messenger  RNA  (mRNA)  which  is 
then  translated  by  reactions  in  the  cell  into  protein.  Each 
gene  contains  the  information  for  a specific  protein. 

SOURCE:  Otlice  of  Technology  Assessment. 


Developing  genetic  technologies 


In  the  each  196()'s.  scientists  disco\ered  e.\- 
actly  how  geties  mo\e  Irom  one  hacterium  to 
atiother.  One  such  mechanism  irses  hacttM'io- 
phages— viruses  that  inlect  bacteria— as  int(M'- 
tuediaries.  F’hages  act  like  In  podertnic  needles, 
injecting  their  1),\.\  into  hactenial  hosts,  where 
it  resides  hel'ore  being  passed  along  from  one 
generation  to  the  next  as  part  of  the  bacterium's 
own  D.\,\.  Sotiietimes,  however,  the  injected 
ON  \ enters  an  activ  e phase  and  produces  a crop 
of  new  V irus  pai'ticles  that  can  then  hurst  out  of 
their  host.  Often  during  this  [jrocess,  the  viral 
0.\.\  inadv  ertently  takes  a piece  of  the  bacterial 


l)\.\  along  w ith  it.  I hus,  vv  Ikmi  the  nrnv  virus 
particUxs  now  infect  other  bacteria,  they  bring 
along  srneral  genes  from  their  pi'evious  host. 
This  viral  transduction— the  transfer  of  genes 
by  an  intei'mediate  viral  vector  or  vehicle— 
could  he  used  to  confer  new  genetic  traits  on 
recipient  bacteria.  (See  figure  12.) 

hactei'ia  also  transfer  genes  directly  in  a proc- 
ess called  conjugation,  in  which  one  hacterium 
attaches  small  projections  to  the  surface  of  a 
nearby  hacterium.  DN',\  from  the  donor  hacte- 
rium is  then  |)assed  to  the  recipient  through  the 


Figure  12.— Transduction:  The  Transfer  of  Genetic  Material  in  Bacteria  by  Means  of  Viruses 


Bacterium 


Bacterial 


In  step  1 of  viral  transduction,  the  infecting  virus  injects  its  DNA  into  the  cell.  In  step  2 when  the  new  viral  particles  are 
formed,  some  of  the  bacterial  chromosomal  fragments,  such  as  gene  A,  may  be  accidently  incorporated  into  these  progeny 
viruses  instead  of  the  viral  DNA.  In  step  3 when  these  particles  infect  a new  cell,  the  genetic  elements  incorporated  from  the 
first  bacterium  can  recombine  with  homologous  segments  in  the  second,  thus  exchanging  gene  A for  gene  a. 


SOURCE:  Office  of  Technology  Assessment. 


40  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


projections.  The  ability  to  form  projections  and 
donate  genes  to  neighbors  is  a genetically  con- 
trolled trait.  The  genes  controlling  this  trait, 
however,  are  not  located  on  the  bacterial  chro- 
mosomes. Instead,  they  are  located  on  separate 
genetic  elements  called  plasmids— relatively 
small  molecules  of  double-stranded  DNA,  ar- 
ranged as  closed  circles  and  existing  autono- 
mously within  the  bacterial  cytoplasm.  (See 
figure  13.) 

Plasmids  and  phages  are  two  vehicles— or 
vectors— for  carrying  genes  into  bacteria.  As 
such,  they  became  tools  of  genetic  engineering; 
for  if  a specifically  selected  DNA  could  he  intro- 
duced into  these  vectors,  it  would  then  he  pos- 
sible to  transfer  into  bacteria  the  hluepi'ints  for 
proteins— the  building  blocks  of  genetic  charac- 
teristics. 

But  bacteria  had  been  confronting  the  inva- 
sion of  foreign  DNA  for  millennia,  and  they  had 
evolved  protective  mechanisms  that  preserved 
their  own  DNA  while  destroying  the  DNA  that 
did  not  belong.  Bacteria  survive  by  producing 
restriction  enzymes.  These  cut  DNA  molecules 
in  places  where  specific  sequences  of  nucleo- 
tides occur— snipping  the  foreign  DNA,  yet  leav- 
ing the  bacteria’s  own  genetic  complement 
alone.  The  first  restriction  enzyme  that  was  iso- 


lated, for  instance,  would  cut  DNA  only  when  it 
located  the  sequence: 

G-A-A-r-r-c 

C-T-T-A-A-G 

If  the  sequence  occurred  once  in  a circular  plas- 
mid, the  effect  would  simply  he  to  open  the 
circle.  If  the  sequence  were  repeated  se\(M’al 
times  along  a length  of  DNA,  the  DN.\  would  he 
chopped  into  se\  eral  small  pieces. 

By  the  late  1970's,  scores  of  different  i’(\stric- 
tion  enzymes  had  been  isolated  fi’om  a \ ai'iety 
of  bacteria,  with  each  enzyme  ha\  ing  a uni(|ue 
specificity  for  one  specific  nucleotide  se(|uence. 
These  enzymes  were  another  key  to  g(‘netic  en- 
gineering: they  not  only  allow  cul  plasmids  to  he 
opened  up  so  that  new  DNA  could  he*  in.serti'd, 
hut  offered  a way  of  obtaining  manageahU* 
pieces  of  new'  DNA  as  w(dl,  (See  figui'e  14.) 
Using  restriction  enzynu\s,  almost  any  DNA 
molecule  could  he  snipped,  shapiul,  and 
trimmed  with  |)recision. 

Cloning  DNA— that  is  obtaining  a large  (|uanti- 
ty  of  exact  copies  of  any  chosen  DNA  molecule 
by  inserting  it  into  a host  bacterium- became 
technically  almost  simpU;.  The  |)iece  in  {|ue.stion 
was  merely  snipped  from  th('  oi’iginal  molecule, 
inserted  into  the  \ ector  DN,\,  and  pro\  ided  w ilh 


Figure  13.— Conjugation:  The  Transfer  of  Genetic  Material  in  Bacteria  by  Mating 


In  conjugation,  a plasmid  inhabiting  a bacterium  can  transfer  the  bacterial  chromosome  to  a second  cell  where  homologous 
segments  of  DNA  can  recombine,  thus  exchanging  gene  B from  the  first  bacterium  for  gene  b from  the  second. 


SOURCE:  Office  of  Technology  Assessment. 


Ch.  2— Introduction  • 41 


Figure  14.— Recombinant  DNA:  The  Technique  of 
Recombining  Genes  From  One  Species 
With  Those  From  Another 


amount  of  DNA  protein 


Restriction  enzymes  recognize  certain  sites  along  the  DNA 
and  can  chemically  cut  the  DNA  at  those  sites.  This  makes 
it  possible  to  remove  selected  genes  from  donor  DNA  mole- 
cules and  insert  them  into  plasmid  DNA  molecules  to  form 
the  recombinant  DNA.  This  recombinant  DNA  can  then  be 
cloned  in  its  bacterial  host  and  large  amounts  of  a desired 
protein  can  be  produced. 

SOURCE;  Office  of  Technology  Assessment. 

a bacterial  host  as  a suitable  en\  ironnient  for 
replication.  The  desired  piece  of  D\,A  could  be 
recombined  \\  ith  a plasmid  \ ector,  a procedure 
that  ga\  e rise  to  recombinant  D.\.A  (rDX.A),  also 
known  as  gene  splicing.  Since  bacteria  can  be 
grown  in  \ast  quantities,  this  process  could 
result  in  large-scale  production  of  otherwise 
scarce  and  e.\pensi\  e proteins. 

.Although  placing  genes  inside  of  bacteria  is 
now  a relati\  ely  straightforward  procedure,  ob- 
taining precisely  the  right  gene  can  be  difficult. 
Three  techniques  are  currently  ax  ailable: 

• Ribonucleic  acid— R\A— is  the  \ehicle 
through  which  the  message  of  D\A  is  read 
and  transcribed  to  form  proteins.  The  Ri\A 
that  carries  the  message  for  the  desired 
protein  is  first  isolated.  An  enzyme,  called 
‘reverse  transcriptase/  is  then  added  to  the 
RNA.  The  enzyme  triggers  the  formation  of 
D\.A— rex  ersing  the  normal  process  of  pro- 
tein production.  The  DNA  is  then  inserted 


into  an  ap|)ro[)riate  \eclor.  This  was  the 
procedure  used  to  obtain  the  gene  for  hu- 
man insulin  in  1979.  (See  figure  15.) 

• The  gene  can  also  he  synthesized,  or 
created,  directly,  since  the  nucleotide  se- 
(|uence  of  the  gene  can  he  deduced  from 
the  amino  acid  seciuence  of  its  protein 
product.  This  procedure  has  worketl  well 
foi'  small  protein.s— like  the  growth  regu- 
latory hormone  somatostatin— which  ha\e 
relatixely  short  stretches  of  DNA  coding. 
Rut  somatostatin  is  a tiny  protein,  only  14 
amino  acids  long.  With  three  nucleotides 
coding  for  each  amino  acid,  scientists  had 
to  synthesize  a DNA  chain  42  nucleotides 
long  to  [)i'otluce  the  coni|)lete  hormone.  For 
larger  proteins,  the  gene-synthesis  ap- 
|)i'oach  rapidly  becomes  highly  impractical. 

• The  third  method  is  also  the  most  con- 
troversial. In  this  "shotgun”  approach,  the 
entire  genetic  complement  of  a cell  is 
chopped  up  by  restriction  enzymes.  Each 
of  the  DNA  fragments  is  attached  next  to 
vectors  and  transferred  into  a bacterium; 
the  bacteria  are  then  screened  to  find  those 
making  the  desired  product.  Screening 
thousands  of  bacterial  cultures  was  part  of 
the  technique  that  enabled  the  isolation  of 
the  human  interferon  gene.* 

At  present,  these  techniques  of  recombina- 
tion work  mainly  with  simple  micro-organisms. 
Scientists  have  only  recently  learned  how  to  in- 
troduce novel  genetic  material  into  cells  of 
higher  plants  and  animals.  These  higher  cells 
are  being  ‘engineered’  in  totally  different  ways, 
by  grow  ing  plant  or  animal  cells  in  ‘tissue  cul- 
ture’ systems,  in  vitro. 

Tissue  culture  systems  work  with  isolated 
cells,  with  entire  pieces  of  tissue,  and  to  a far 
more  limited  extent,  with  whole  organs  or  ev  en 
early  embryos.  The  techniques  make  it  possible 
to  manipulate  cells  experimentally  and  under 
controlled  conditions.  Several  techniques  are 
available.  For  example,  in  one  set  of  experi- 
ments, complete  plants  have  been  grown  from 
single  cells— a breakthrough  that  may  permit 


'Strictly  speaking,  R\A  was  transcribed  using  the  shotgun 
approach  into  DNA,  which  was  then  cloned  into  bacteria  and 
screened. 


42  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Fiqure  15.— An  Example  of  How  the  Recombinant  DNA  Technique  May  Be  Used 
^ To  Insert  New  Genes  Into  Bacterial  Cells 


The  first  part  of  the  technique  invoives  the  manipuia- 
tions  necessary  to  isoiate  and  reconstruct  the  desired 
gene  from  the  donor: 

a)  The  RNA  that  carries  the  message  (mRNA)  for  the 
desired  protein  product  is  isoiated. 

b)  The  doubie-stranded  DNA  is  reconstructed  from  the 
mRNA. 

c)  In  the  finai  step  of  this  sequence,  the  enzyme  ter- 
minai  transferase  acts  to  extend  the  ends  of  the 
DNA  strands  with  short  sequences  of  identical 
bases  (in  this  case  four  guanines). 


a) 


b) 


I 

Messenger  RNA 
from  animal  cell 


Double-strand  DNA 


Enzymatic 

reconstruction 


c) 


Terminai 

transferase 


GGG(5i 


III.  The  final  product,  a bacterial  plasmid  containing  the 
new  gone,  is  obtained.  This  piasmid  can  then  be  in- 
serted into  a bacterium  where  it  can  be  repiicated  and 
produce  the  desired  protein  product: 

a)  The  gene  obtained  in  part  I and  the  plasmid  DNA 
from  part  II  are  mixed  together  and  anneal 
because  of  the  complementary  base-pairing  be- 
tween them. 

b)  Bacterial  enzymes  fill  in  any  gaps  in  the  circle, 
sealing  the  connection  between  the  plasmid  DNA 
and  the  inserted  DNA  to  generate  an  intact  cir- 
cular plasmid  now  containing  a new  gene. 


II.  A bacterial  plasmid,  which  is  a small  piece  of  circular 
DNA,  serves  as  the  vehicle  for  introducing  the  new  gene 
(obtained  in  part  I above)  into  the  bacterium: 

a)  The  circular  plasmid  is  cleaved  by  the  appropriate 
restriction  enzyme. 

b)  The  enzyme  terminal  transferase  extends  the  DNA 
strands  of  the  broken  circle  with  identical  bases 
(four  cytosines  in  this  case,  to  allow  complemen- 
tary base  pairing  with  the  guanines  added  to  the 
gene  obtained  in  part  I). 

II 


Bacterial  plasmid  DNA 


Uptake  by  cell; 
repair  by 


SOURCE:  Office  of  Technology  Assessment. 


Ch.  2— Introduction  • 43 


luinclreds  ot  |)lants  lo  he  ^rown  asexiialK’  from 
a small  sam|)le  of  plant  material.  Just  as  with 
tiaeteria,  the  cells  can  he  itKlucecI  lo  lake  u|) 
pieces  of  n\'A  in  a process  called  transforma- 
tion. rhey  can  also  he  e.xposed  to  mutation- 
causing  agents  so  that  they  produce  mutants 
with  desired  propei'ties.  In  another  set  of  exper- 
iments, two  different  cells  ha\e  been  fused  to 
form  a new,  single-cell  “Inhrid”  that  contains 
the  genetic  complements  of  both  antecedents. 
In  both  cases,  the  success  of  tissue  culture  and 


The  basic  issues  

.Applied  genetics  is  like  no  other  lechnologx’. 
B\  itself,  it  may  enable  ti’emendous  ad\  ances  in 
conc|uering  diseases,  increasing  food  pi'oduc- 
tion,  producing  new  and  cheaper  industrial  sub- 
stances, cleaning  up  pollution,  and  understand- 
ing the  fundamental  processes  of  life.  B('cause 
the  technologN’  is  so  |K)\\  erful,  and  because  it  in- 
\ ol\  es  the  basic  loots  of  life  itself,  it  carries  w ith 
it  potential  hazards,  some  of  w hich  might  arise 
from  basic  research,  others  of  w hich  ma\'  stem 
from  its  applications. 

As  the  impacts  of  genetic  technologies  are  dis- 
cussed, two  fundamental  (]uestions  must  he 
kept  in  mind: 

How  will  applied  genetics  be  used? 

Interest  in  the  industrial  use  of  biological 
processes  stems  from  a merging  of  two  paths: 
the  re\  olution  in  scientific  understanding  of  the 
nature  of  genetics:  and  the  accelerated  search 
for  a sustainable  society  in  which  most  indus- 
trial processes  are  based  on  the  use  of  renew- 
able resources.  The  new  genetic  technologies 
will  spur  that  search  in  three  ways:  they  will 
pro\  ide  a means  of  doing  something  biolog- 
ically—with  renewable  raw  materials — that  pre- 
\ iously  required  chemical  processes  using  non- 
renewable  resources;  they  will  offer  more  ef- 
ficient, more  economical,  less  polluting  ways  for 
producing  both  old  and  new  products;  and  they 
will  increase  the  yield  of  the  plant  and  animal 
resources  that  are  responsible  for  providing  the 
world's  supplies  of  food,  fibers,  and  some  fuels. 


cell  fusion*  can  he  used  to  direct  efficient,  fast 
genetic  changes  in  plants.  (See  ch.  8.) 

(!ell  culture  lechni(|ues,  while  not  sti'ictly  g(v 
netic  manipulation,  form  a majoi’  aspi'ct  ot  mod- 
ern biotechnology,  ('omhined  with  genetic  ap- 
proaches, their  |)otential  is  only  on  th(^  \ (M'g(^  ot 
being  realized. 

'.A  related  leehni(|ue  is  protoplast  fusioo,  or  the  fusion  ol  cells 
whose  walls  have  been  renun ed  to  leave  only  minuhrane-hoiind 
cells.  The  cells  of  hacteria.  funf'i.  and  plants  must  all  he  freed  of 
their  walls  Itefore  they  can  he  fused. 


ll'hat  are  the  dangers? 

K\en  before  scicMitists  recognized  the  jjoten- 
tial  power  of  applied  genetics,  some  c|uestioned 
its  conseciuences;  for  w ith  its  benefits,  ap[)eared 
hypothetical  risks.  .Although  most  exptfrts  today 
agree  that  the  immediate  hazards  of  the  basic 
research  itself  appear  to  he  minimal,  nobody 
can  he  certain  about  all  the  conseciuences  of 
placing  genetic  characteristics  in  micro-orga- 
nisms, plants,  and  animals  that  ha\e  nev  er  car- 
ried them  before.  There  are  at  least  three  sepa- 
rate areas  of  concern: 

First,  genetically  engineered  micro-organisms 
might  have  potentially  deleterious  effects  on  hu- 
man health,  other  living  organisms,  or  the  envi- 
ronment in  general.  Unlike  toxic  chemicals,  or- 
ganisms may  reproduce  and  spread  of  their 
own  accord:  if  they  are  released  into  the  envi- 
ronment, they  may  be  impossible  to  control. 

Second,  some  observers  c[uestion  whether 
sufficient  knowledge  exists  to  allow'  the  extinc- 
tion of  diverse  species  of  “genetically  inferior’’ 
plants  and  animals  in  favor  of  a few  strains  of 
"superior”  ones.  Evolution  thus  far  has  de- 
pended, in  part,  on  genetic  diversity;  replacing 
in  nature  div  erse  inferior  strains  by  genetically 
engineered  superior  strains  may  increase  the 
susceptibility  of  living  things  to  disease  and  en- 
vironmental insults. 

Finally,  this  new  knowledge  affects  the  un- 
derstanding of  life  itself.  It  is  tied  to  the  ultimate 


44  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


c|uestions  of  how  humans  view  themselves  and 
what  they  legitimately  control  in  the  world. 

Because  of  the  significant  and  wide-ranging 
scope  of  applied  genetics,  society  as  whole  must 
begin  to  debate  the  issues  with  a view  toward  al- 


locating and  monitoring  its  benefits  and  bur- 
dens. That  process  requires  knowledge.  The  fol- 
lowing sections  of  the  report  describe  the  im- 
pacts of  applied  genetics  on  specific  industries, 
and  assess  many  of  their  consequences. 


Part  I 


Biotechnology 


! Chapter  3— Genetic  Engineering  in  the  Fermentation  Technologies 49 

I Chapter  4— The  Pharmaceutical  Industry 59 

i Chapter  5— The  Chemical  Industry 85 

' Chapter  6— The  Food  Processing  Industry 107 

i Chapter  7— The  Use  of  Genetically  Engineered  Micro-Organisms  in  the  Environment  . . 117 


I 

I 


chapter  3 

Genetic  Engineering  and  the 
Fermentation  Technologies 


chapter  3 


Page 


Biotechnology— An  Introduction 49 

Fermentation 49 

Fermentation  Industries 50 

Fermentation  Using  Whole  Living  Cells 51 

The  Process  of  Enzyme  Technology 53 

Comparative  Advantages  of  Fermentations 
Using  Whole  Cells  and  Isolated  Enzymes ....  54 
The  Relationship  of  Genetics  to  Fermentation  . 54 
Fermentation  and  Industry 55 


Figures 


Figure  No.  Page 

16.  Diagram  of  Products  Available  From  Cells  ...  50 

17.  Features  of  a Standard  Fermenter 52 

18.  Immobilized  Cell  System 52 

19.  Diagram  of  Conversion  of  Raw  Material 

to  Product 53 


Table 

Table  No. 

2.  Enzyme  Products 


Page 

54 


Chapter  3 

Genetic  Engineering  and 
the  Fermentation  Technologies 

Biotechnology — an  introduction  


Biotechnology  imoKes  the  use  in  industry  of 
Ii\  ing  organisms  or  tlieir  compotients  (such  as 
enzymes).  It  includes  the  introduction  of  geneti- 
calh'  engineered  micro-organisms  into  a \ ariety 
of  industrial  [)rocesses. 

rhe  [)harmaceutical,  chemical,  and  food  proc- 
essing industries,  in  that  order,  are  most  likely 
to  take  ad\  antage  of  ad\  ances  in  molecular  ge- 
netics. Others  that  might  also  he  affected,  al- 
though not  as  immediateh',  are  the  mining, 
crude  oil  recoxery,  and  pollution  control  in- 
dustries. 

Because  nearly  all  the  products  of  hiotechnol- 
og\'  are  manufactured  hy  micro-organisms,  fer- 
mentation is  an  indispensihle  element  of  hio- 
technology's  suppoi't  system.  'I'he  pharmaceuti- 
cal industry,  the  earliest  beneficiary  of  the  new 
knowledge,  is  already  producing  pharmaceu- 
ticals derixed  from  geneticallx'  engineei'ed 
micro-organisms.  The  chemical  industry  xx  ill 
take  longer  to  make  use  of  biotechnology,  hut 
the  ultimate  impact  max’  he  enormous.  The  food 
processing  industry  xxill  probably  he  affected 
last. 

This  report  e.xamines  many  of  the  pharma- 
ceutical industry’s  products  in  detail,  as  xxell  as 


Fermentation  

There  are  sex  eral  xx  ays  that  D\A  can  be  cut, 
spliced,  or  otherwise  altered.  But  engineered 
D\,A  by  itself  is  a static  molecule.  To  be  any- 
thing more  than  the  end  of  a laboratory  exer- 
cise, the  molecule  must  be  integrated  into  a sys- 
tem of  production;  to  hax  e an  impact  on  society 
at  large,  it  must  become  a component  of  an  in- 
dustrial or  otherxvise  useful  process. 

The  process  that  is  central  to  the  economic 


some  of  the  secondary  impacts  that  the  technol- 
ogies might  haxe.  Because  the  chemical  and 
food  industries  xx  ill  feel  the  major  impact  of  bio- 
technology later,  specific  impacts  are  less  cer- 
tain and  particular  pi'oducts  ai’e  less  identifi- 
able. The  mining,  oil  recoxery,  and  pollution 
control  industries  are  also  candidates  for  the 
use  of  genetic  technologies.  Hoxx  ex  er,  because 
of  technical,  scientific,  legal,  and  economic  un- 
certainties, the  success  of  apjjlications  in  these 
industries  is  more  speculatix  e. 

The  generalizations  made  xvith  respect  to 
each  of  the  industries  should  be  x iexxed  as  just 
that— generalizations.  Because  a xvide  array  of 
products  can  be  made  biologically,  and  because 
different  factors  influence  each  instance  of  pro- 
duction, isolated  examples  of  success  may  ap- 
pear throughout  the  industries  at  approximate- 
ly the  same  time.  In  almost  ex'ery  case,  specific 
predictions  can  only  be  made  on  a product-by- 
product basis;  for  xx  hile  it  may  be  true  that  bio- 
technologx'’s  oxerall  impact  will  be  profound, 
identifying  many  of  the  products  most  likely  to 
be  affected  remains  speculatix  e. 


success  of  biotechnology  has  been  around  for 
centuries.  It  is  fermentation,  essentially  the 
process  used  to  make  xvine  and  beer.  It  can  also 
produce  organic  chemical  compounds  using 
micro-organisms  or  their  enzymes. 

Ox  er  the  years,  the  scope  and  efficiency  of 
the  fermentation  process  has  been  gradually  im- 
proxed  and  refined.  Txvo  processes  now  exist, 
both  of  xvhich  xvill  beneft  from  genetic  engi- 


49 


50  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


neering.  In  fermentation  technology,  living  or- 
ganisms serve  as  miniature  factories,  convert- 
ing raw  materials  into  end  products.  In  enzyme 
technology,  biological  catalysts  extracted  from 
those  living  organisms  are  used  to  make  the 
products. 

Fermentation  industries 

The  food  processing,  chemical,  and  pharma- 
ceutical industries  are  the  three  major  users  of 
fermentation  today.  The  food  industry  was  the 
first  to  exploit  micro-organisms  to  produce 
alcoholic  beverages  and  fermented  foods.  Mid- 
16th  century  records  describe  highly  sophisti- 
cated methods  of  fermentation  technology.  Heat 
processing  techniques,  for  example,  anticipated 
pasteurization  by  several  centuries. 

In  the  early  20th  century,  the  chemical  in- 
dustry began  to  use  the  technology  to  produce 
organic  solvents  like  ethanol,  and  enzymes  like 
amylase,  used  at  the  time  to  treat  textiles.  The 
chemical  industry’s  interest  in  fermentation 
arose  as  the  field  of  biochemistry  took  shape 
around  the  turn  of  the  century.  But  it  was  not 
until  World  War  I that  wartime  needs  for  the 
organic  solvent  acetone— to  produce  the  cor- 
dite used  in  explosives— substantially  increased 
research  into  the  potential  of  fermentation. 
Thirty  years  later  after  World  War  II,  the  phar- 
maceutical industry  followed  the  chemical  in- 
dustry’s lead,  applying  fermentation  to  the  pro- 
duction of  vitamins  and  new  antibiotics. 

Today,  approximately  200  companies  in  the 
United  States  and  over  500  worldwide  use 
fermentation  technologies  to  produce  a wide 
variety  of  products.  Most  use  them  as  part  of 
production  processes,  usually  in  food  process- 
ing. But  others  manufacture  either  proteins, 
which  can  be  considered  primary  products,  or  a 
host  of  secondary  products,  which  these  pro- 
teins help  produce.  For  genes  can  make  en- 
zymes, which  are  proteins;  and  the  enzymes 
can  help  make  alcohol,  methane,  antibiotics, 
and  many  other  substances. 

Proteins,  the  primary  products,  function  as: 

• enzymes  such  as  asparaginase  which  are 
used  in  the  treatment  of  leukemia; 


• structural  components,  such  as  collagen, 
used  in  skin  transplants  following  burn 
trauma; 

• certain  hormones,  such  as  insulin  and 
human  growth  hormone; 

• substances  in  the  immune  system,  such  as 
antibodies  and  interferon:  and 

• specialized  functional  components,  such  as 
hemoglobin. 

Fermentation  technologies  are  so  useful  for  pro- 
ducing proteins  partly  because  these  are  the 
direct  products  of  genes.  But  proteins  (as  en- 
zymes) can  also  be  used  in  thousands  of  addi- 
tional conversions  to  produce  practically  any 
organic  chemical  and  many  inorganic  ones  as 
well:  (See  figure  16.) 


Figure  16.— Diagram  of  Products  Available 
From  Cells 


In  (A)  DNA  directs  the  formation  of  a protein,  such  as  in- 
sulin, which  is  itself  the  desired  product.  In  (B),  DNA  directs 
the  formation  of  an  enzyme  which,  in  turn,  converts  some 
raw  material,  such  as  sugar,  to  a product,  such  as  ethanol. 

SOURCE:  Office  of  Technology  Assessment. 


Ch.  3— Genetic  Engineering  and  the  Fermentation  Technologies  • 51 


• rai'holndrates,  such  as  fructose  sweeten- 
ei's: 

• lipids,  such  as  \ itamins  A,  E,  and  K; 

• alcohols,  such  as  ethanol; 

• other  oi'ganic  compounds,  such  as  acetone: 
and 

• inorganic  chemicals,  such  as  ammonia,  for 
use  in  fertilizers. 

Fermentation  is  not  the  onh'  \\a\  to  manufac- 
ture or  isolate  these  products.  Some  are  tradi- 
tionalK  produced  hy  other  methods.  If  a change 
from  one  pi'oeess  to  anothei'  is  to  occur,  both 
economic  and  societal  pressures  \\  ill  help  deter- 
mine whether  an  inno\ati\e  a[)proach  will  he 
used  to  [)i'oduce  a [)ai  ticular  product.  .Alan  Bull 
has  identified  four  stimuli  for  change  and  in- 
no\  ation:' 

1.  abundance  of  a [jotentially  useful  raw 
material: 

2.  scarcitx’  of  an  established  product; 

3.  disco\  ery  of  a new  product:  and 

4.  en\  ironmental  concei'iis. 

.And  conditions  e.xisting  toda\  ha\  e added  a fifth 
stimulus: 

5.  scarcity  of  a currently  used  raw  material. 

Each  of  these  factors  has  tended  to  accelerate 
the  application  of  fermentation. 

1.  Abundance  of  a potentially  useful  raw  ma- 
terial.—The  use  of  a raw  material  can  be 
the  dri\  ing  force  in  dex  eloping  a process. 
When  straight  chain  hydrocarbons  (n-al- 
kanes)  were  produced  on  a large  scale  as 
petroleum  refinery  byproducts,  fermenta- 
tion processes  were  developed  to  conxert 
them  to  single-cell  proteins  for  use  in  ani- 
mal feed. 

2.  Scarcity  of  an  established  product.— The 
new-found  potential  for  producing  human 
hormones  through  fermentation  technol- 
og\'  is  a major  impetus  to  the  industry  to- 
day. Similarly,  many  organic  compounds 
once  obtained  by  other  processes— like 
citric  acid,  which  was  extracted  directly 


'.A.  T.  Bull,  D.  C.  Elluood,  and  C.  Ralledge,  Microbial  Technology: 
Current  State,  Future  Prospects,  29th  Symposium  of  the  Society  tor 
(ieneral  .Microbiologx'  at  University  of  Cambridge.  .April  1979 
(Cambridge.  England:  Cambridge  University  Press.  1979).  pp.  4-8. 


from  citrus  fruits— are  now  made  hy  fer- 
mentation. .As  a result  of  more  efficient 
technology,  pi’oducts  from  \itamin  B,,  to 
steroids  ha\  e come  into  w ider  use. 

3.  Discovery  of  a new  product.— The  discox  erv 
that  antibiotics  were  produced  hy  micro- 
organisms sparked  searches  for  an  entirely 
new  group  of  jii'otlucts.  Several  thousand 
antibiotics  have  been  discovered  to  date,  of 
w hich  over  a hundred  have  proved  to  be 
clinically  useful. 

4.  Environmental  concerns.— I'he  problems  of 
sewage  treatment  and  tbe  need  for  new 
sources  of  energy  have  triggered  a search 
foi'  methods  to  convert  sewage  and  munici- 
pal wastes  to  methane,  the  principal  com- 
ponent of  natural  gas.  Because  micro-orga- 
nisms play  a major  role  in  the  natural  cy- 
cling of  organic  compounds,  fermentation 
has  been  one  method  usetl  for  the  conver- 
sion. 

5.  Scarcity  of  a currently  used  raw  materi- 
al.—Because  the  Earth’s  supplies  of  fossil 
fuels  are  rapidly  dwindling,  there  is  intense 
interest  in  finding  methods  for  converting 
other  raw  materials  to  fuel.  Fermentation 
offers  a major  approach  to  such  conver- 
sions. 

Fermentation  technologies  can  be  effective  in 
each  of  these  situations  because  of  their  out- 
standing versatility  and  relative  simplicity.  The 
processes  of  fermentation  are  basically  identi- 
cal, no  matter  what  organism  is  selected,  what 
medium  used,  or  what  product  formed.  The 
same  apparatus,  with  minor  modifications,  can 
be  used  to  produce  a drug,  an  agricultural  prod- 
uct, a chemical,  or  an  animal  feed  supplement. 

Fermentation  using  whole  living  cells 

Originally,  fermentation  used  some  of  the 
most  primitive  forms  of  plant  life  as  cell  fac- 
tories. Bacteria  were  used  to  make  yogurt  and 
antibiotics,  yeasts  to  ferment  wine,  and  the 
filamentous  fungi  or  molds  to  produce  organic 
acids.  More  recently,  fermentation  technology 
has  begun  to  use  cells  derived  from  higher 
plants  and  animals  under  growdh  conditions 
known  as  cell  or  tissue  culture.  In  all  cases, 
large  quantities  of  cells  with  uniform  character- 


52  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


islics  are  grown  under  defined,  controlled  con- 
ditions. 

In  its  simplest  form,  fermentation  consists  of 
mixing  a micro-organism  with  a liquid  broth 
and  allowing  the  components  to  react.  More  so- 
phisticated large-scale  processes  require  control 
of  the  entire  environment  so  that  fermentation 
proceeds  efficiently  and,  more  importantly,  so 
that  it  can  be  repeated  exactly,  with  the  same 
amounts  of  raw  materials,  broth,  and  micro- 
organisms producing  the  same  amount  of  prod- 
uct. Strict  control  is  maintained  of  such  vari- 
ables as  pH  (acidity/alkalinity),  temperature,  and 
oxygen  supply.  (See  figure  17.)  The  newest  mod- 
els are  regulated  by  sensors  that  are  monitored 
by  computers.  The  capacity  of  industrial-sized 
fermenters  can  reach  50,000  gal  or  more.  The 
one-shot  system  of  fermentation  is  called  batch 
fermentation— i.e.,  fermentation  in  which  a 
single  batch  of  material  is  processed  from  start 
to  finish. 

In  continuous  fermentation,  an  improvement 
on  the  batch  process,  fermentation  goes  on 
without  interruption,  with  a constant  input  of 

Figure  17.— Features  of  a Standard  Fermenter 

Exhaust 


raw  materials  and  other  nutrients  and  an  at- 
tendant output  of  fermented  material.  The  most 
recent  approaches  use  micro-organisms  that 
have  been  immobilized  in  a supporting  struc- 
ture. (See  figure  18.)  As  the  solution  containing 
the  raw  material  passes  over  the  cells,  the 
micro-organisms  process  the  material  and  re- 
lease the  products  into  the  solution  flowing  out 
of  the  fermenter. 

In  general,  products  obtained  by  fermenta- 
tion also  can  be  produced  by  chemical  synthe- 
sis, and  to  a lesser  extent  can  he  isolated  by  ex- 
traction from  whole  organs  or  oi'ganisms.  A 
fermentation  process  is  usually  most  competi- 
tive when  the  chemical  process  retjuires  sex  eral 

Figure  18.— Immobilized  Cell  System 


Solution  with  product  out 

f ■ 


) 

Raw  material  solution  in 


Typically,  a solution  of  raw  materials  is  pumped  through  a 
bed  of  immobilized  micro-organisms  which  convert  the 
materials  to  the  desired  product. 

SOURCE:  Office  of  Technology  AssessmenI 


Ch.  3— Genetic  Engineering  and  the  Fermentation  Technologies  • 53 


incli\  iilual  steps  to  c'omj)lete  the  com  ersion.  In  a 
cliemical  synthesis,  the  I'aw  mattM’ial  (shown  in 
t'if^ure  19  as  a)  might  have  to  he  transtbnned  to 
an  intermediate  h.  w liich,  in  tiii’ii,  might  lia\  e to 
he  comerted  to  intermediates  c and  d het'ore 
final  comei'sion  to  the  [)rodiict  e— eacli  step 
necessitating  the  recovery  of  its  products  before 
tlie  next  con\ersion.  In  fei'mentation  technol- 
og\',  all  steps  take  place  within  those  miniature 
chemical  factories,  the  micro-oi'ganisms;  the 
microbial  chemist  merely  adds  the  I'aw  material 
a and  reco\  ers  the  pioduct  e. 

A v\  ide  \ ariety  of  cai'holndrate  raw  materials 
can  be  used  in  fermentation.  These  can  he  pure 
substances  (sucrose  or  table  sugar,  glucose,  or 
fructose)  or  complex  mixtures  still  in  their 
original  form  (cornstalks,  potato  mash,  sugar- 
cane, sugar  beets,  orcellulose).  They  can  he  of 
recent  biological  origin  (biomass)  oi'  derived 
from  fossil  fuels  (methane  or  oil).  The  availabili- 
ty of  raw'  materials  varies  from  country  to  coun- 


Figure  19.— Diagram  of  Conversion  of 
Raw  Material  to  Product 


a)  Chemical  conversion 


a 

-►b 

V 



-►d 

J 

►e 

Raw 

material 

Intermediate 

products 

Final 

product 

b)  Biological  conversion 


material  product 


3)  In  the  chemical  conversion  of  raw  material  a to  final 
product  e,  intermediates  b,  c,  and  d must  be  synthe- 
sized. Each  intermediate  must  be  recovered  and  purified 
before  it  can  be  used  in  the  next  step  of  the  conversion. 

b)  A cell  can  perform  the  same  conversion  of  a to  e,  but 
with  the  advantage  that  the  chemist  does  not  have  to 
deal  with  the  intermediates:  the  raw  material  a is  simply 
added  and  the  final  product  e,  recovered. 


SOURCE:  Office  of  Technology  Assessment. 


try  and  even  from  region  to  region  within  a 
country;  the  economics  r>f  the  production  proc- 
ess varies  accordingly. 

The  cost  of  the  raw  material  can  contribute 
significantly  to  the  cost  of  [troduction.  Usually, 
the  most  useful  micro-organisms  are  those  that 
consume  reatlily  available  inexpensive  raw'  ma- 
terials. For  large  volume,  low-priced  products 
(such  as  commodity  chemicals),  the  relationshi|) 
between  the  cost  of  the  i'aw  material  and  the 
cost  of  the  end  product  is  significant.  For  low 
volume,  high-priced  products  (such  as  certain 
pharmaceuticals),  the  relationship  is  negligible. 


The  process  of  enzyme  technology 


.Although  live  yeast  had  been  used  for  several 
thousand  years  in  the  production  of  fermented 
foods  and  beverages,  it  was  not  until  1878  that 
the  active  agents  of  the  fermentation  process 
were  given  the  name  "enzymes”  (from  the 
Greek,  meaning  "in  yeast”).  The  inanimate 
nature  of  enzymes  was  demonstrated  less  than 
two  decades  later  when  it  was  shown  that  ex- 
tracts from  yeast  cells  could  effect  the  conver- 
sion of  glucose  to  ethanol.  Finally,  their  actual 
chemical  nature  was  established  in  1926  with 
the  purification  and  crystallization  of  the 
enzvme  urease. 


Fermentation  carried  out  by  live  cells  pro- 
vided the  conceptual  basis  for  designing  fer- 
mentation processes  based  on  isolated  enzymes. 
A single  enzyme  situated  within  a living  cell  is 
needed  to  convert  a raw  material  into  a prod- 
uct. A lactose-fermenting  organism,  e.g.,  can  be 
used  to  convert  the  sugar  lactose,  which  is 
found  in  milk,  to  glucose  (and  galactose).  But  if 
the  actual  enzyme  responsible  for  the  conver- 
sion is  identified,  it  can  be  extracted  from  the 
cell  and  used  in  place  of  a living  cell.  The 
purified  enzyme  carries  out  the  same  conver- 
sion as  the  cell,  breaking  down  the  raw  material 
in  the  absence  of  any  viable  micro-organism.  An 
enzyme  that  acts  inside  a cell  to  convert  a raw 
material  to  a product  can  also  do  this  outside  of 
the  cell. 

Both  batch  and  continuous  methods  are  used 
in  enzyme  technology.  However,  in  the  batch 
method,  the  enzymes  cannot  be  recovered  eco- 


i'if 


54  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


nomically,  and  new  enzymes  must  be  added  for 
each  production  cycle.  Furthermore,  the  en- 
zymes are  difficult  to  separate  from  the  end 
product  and  constitute  a potential  contaminant. 
Because  enzymes  used  in  the  continuous  meth- 
od are  reusable  and  tend  not  to  be  found  in  the 
product,  the  continuous  method  is  the  method 
of  choice  for  most  processes.  Depending  on  the 
desired  conversion,  the  immobilized  micro- 
organisms of  figure  18  could  be  replaced  by  an 
appropriate  immobilized  enzyme. 

Although  more  than  2,000  enzymes  have 
been  discovered,  fewer  than  50  are  currently  of 
industrial  importance.  Nevertheless,  two  major 
features  of  enzymes  make  them  so  desirable: 
their  specificity  and  their  ability  to  operate 
under  relatively  mild  conditions  of  temperature 
and  pressure.  (The  most  frequently  used  en- 
zymes are  listed  in  table  2.) 

Comparative  advantages  of 
fermentations  using  whole  cells 
and  isolated  enzymes 

At  present,  it  is  still  uncertain  whether  the 
use  of  whole  cells  or  isolated  enzymes  will  be 
more  useful  in  the  long  run.  There  are  advan- 
tages and  disadvantages  to  each.  The  role  of  ge- 
netic engineering  in  the  future  of  the  industry. 


Table  2.— Enzyme  Products 


Source/name 

Commercially 
available  before: 

Current 

production 

tons/yr 

1900 

1950 

1980 

Animal 

Rennet 

X 

2 

Trypsin 

X 

15 

Pepsin 

X 

5 

Plant 

Malt  amylase 

X 

10,000 

Papain 

X 

100 

Microbial 

Koji 

X 

? 

Fungal  protease 

X 

10 

Bacillus  protease  .... 

X 

500 

Amyloglucosidase  . . . 

X 

300 

Fungal  amylase 

X 

10 

Bacterial  amylase .... 

X 

300 

Pectinase 

X 

10 

Glucose  isomerase.  . . 

X 

50 

Microbial  rennet 

X 

10 

however,  will  be  partly  determined  by  which 
method  is  chosen.  With  isolated  enzymes,  ge- 
netic manipulation  can  readily  increase  the  sup- 
ply of  enzymes,  while  with  whole  organisms,  a 
wide  variety  of  manipulations  is  possible  in  con- 
structing more  productive  strains. 

The  relationship  of  genetics 
to  fermentation 

Applied  genetics  is  intimately  tied  to  fermen- 
tation technology,  since  finding  a suitable  spe- 
cies of  micro-organism  is  usually  the  first  step  in 
developing  a fermentation  technique.  Until  re- 
cently, geneticists  have  had  to  search  for  an 
organism  that  already  produced  the  needed 
product.  However,  through  genetic  manipula- 
tion a totally  new  capability  can  be  engineered; 
micro-organisms  can  be  made  to  produce  sub- 
stances beyond  their  natural  capacities.  The 
most  striking  successes  have  been  in  the  phar- 
maceutical industry,  where  human  genes  have 
been  transferred  to  bacteria  to  produce  insulin, 
growth  hormone,  interferon,  thymosin  a-1,  and 
somatostatin.  (See  ch.  4.) 

In  general,  once  a species  is  found,  coinen- 
tional  methods  have  been  used  to  intluce  muta- 
tions that  can  produce  even  more  of  the  d(\sired 
compound.  The  geneticist  searches  fi-om  among 
hundreds  of  mutants  for  the  one  micro-orga- 
nism that  produces  most  efficiently.  Most  of  th(’ 
many  methods  at  the  microbiologist’s  disposal 
involve  trial-and-error.  Newer  g(Mi(!ti('  t('('h- 
nologies,  such  as  the  use  of  recombinant  DNA 
(rDNA),  allow  approaches  in  which  us(’ful  genet- 
ic traits  can  be  inserted  dir(u;lly  into  the?  micro- 
organism. 

The  current  industrial  approach  to  lermenta- 
tion  technologies  therefore  consid(>i's  two  prob- 
lems: First,  whether  a biological  process  can 
produce  a particular  product:  and  second,  w hat 
micro-organism  has  the  gr(;aU‘st  potential  lor 
production  and  how  the  cUisircnl  characlei  islies 
can  be  engineered  for  it.  Finding  the  desii-ed 
micro-organism  and  improving  its  capability  is 
so  fundamental  to  the  lernu'ntation  industry, 
that  geneticists  have  hec'onu^  im|)oi  tant  mem- 
bers of  fermentation  i'(!S(Nirch  teams. 


SOURCE:  Office  of  Technology  Assessmerrt. 


Ch.  3— Genetic  Engineering  and  the  Fermentation  Technologies  • 55 


(lenetic  engineei'ing  can  increase  an  orga- 
nism's proclnetixe  eapal)ility  (a  change  that  can 
make  a process  economically  competitix  e);  hot  it 
can  also  be  used  to  construct  sti'ains  w ith  char- 
acteristics other  than  higher  [)roclucli\  ity.  Prop- 
erties such  as  objectionable  coloi',  odor,  or  slime 
can  he  I'emoved.  Ihe  formation  of  spores  that 
could  lead  to  airborne  spread  of  the  micro- 
organism can  he  su[)pressed.  The  formation  of 
harmful  hv})roducts  can  he  eliminated  oi'  re- 
duced. Other  pi’opei'ties,  such  as  I’esistance  to 
bacterial  \ iruses  and  increased  genetic  stability, 
can  he  gix  en  to  micro-organisms  that  lack  them. 

.-\p[)lying  recent  genetic  engineering  tech- 
niques to  the  production  of  industrially  \ aluahle 
enzymes  may  also  prove  useful  in  the  tuture. 
For  e.xample,  a strain  of  micro-organism  that 
carries  the  genes  for  a desired  enzyme  may  he 
pathogenic.  If  the  genes  that  e.xpress  ([jroduce) 
the  enzyme  can  he  transferred  to  an  innocuous 
micro-organism,  the  enzyme  can  he  produced 
safely. 

Cl  RREiN'T  TECH.MC.XL  LIVRTS  (),\ 
GENETIC  ENGINEERING 

Despite  the  many  genetic  manipulations  that 
are  theoretically  possible,  there  are  several 
notable  technical  limitations; 

• Genetic  maps— the  identification  of  the  lo- 
cation of  desired  genes  on  various  chromo- 
somes have  not  been  constructed  for  most 
industrially  useful  micro-organisms. 

• Genetic  systems  for  industrially  useful 
micro-organisms,  such  as  the  availability  of 
useful  vectors,  are  at  an  early  stage  of 
development. 

• Physiological  pathways— the  sequence  of 
enzymatic  steps  leading  from  a raw  mate- 
rial to  the  desired  product,  are  not  known 
for  many  chemicals.  Much  basic  research 
will  be  necessary  to  identify  all  the  steps. 
The  number  of  genes  necessary  for  the  con- 
version is  a major  limitation.  Currently, 
rDNA  is  most  useful  when  only  a single 
easily  identifiable  gene  is  needed.  It  is  more 
difficult  to  use  when  several  genes  must  be 
transferred.  Finally,  the  problems  are  for- 
midable, if  not  impossible,  when  the  genes 
have  not  yet  been  identified.  This  is  the 


case  with  many  traits  of  agronomic  impor- 
tance, such  as  plant  height. 

Fven  if  the  genes  are  identified  and  suc- 
cessfully transferred,  methods  must  he  de- 
veloped to  recognize  the  bacteria  that  re- 
ceived them.  Fhei'efore,  the  need  to  devel- 
op appropi'iate  selection  methods  has  im- 
peded the  application  of  molecular  ge- 
netics. 

,\s  a conse(|uence  of  these  limitations,  genetic 
engineei'ing  will  he  applied  to  the  development 
of  capabilities  that  re(|uii’e  the  transfer  of  only 
one  or  a few  identified  genes. 

Fermentation  and  industry 

Genetic  engineei'ing  is  not  in  itself  an  indus- 
try, but  a technology  used  at  the  laboratory 
level.  It  allows  the  researcher  to  alter  the  hered- 
itary apparatus  of  a living  cell  so  that  the  cell 
can  produce  more  or  diffei’ent  chemicals,  or 
perform  completely  new  functions.  Fhe  altered 
cell,  or  more  appropriately  the  population  of 
altered  identical  cells  is,  in  turn,  used  in  indus- 
trial production.  It  is  within  this  framework  that 
the  impacts  of  applied  genetics  in  the  various  in- 
dustries is  examined. 

Regardless  of  the  industry,  the  same  three 
criteria  must  be  met  before  genetic  technologies 
can  become  commercially  feasible.  These  cri- 
teria represent  major  constraints  that  industry 
must  overcome  before  genetic  engineering  can 
play  a part  in  bringing  a product  to  market. 
They  include  the  need  for: 

1.  a useful  biochemical  product; 

2.  a useful  biological  fermentation  approach 
to  commercial  production;  and 

3.  a useful  genetic  approach  to  increase  the 
efficiency  of  production. 

The  three  criteria  interrelate  and  can  be  met 
in  any  order;  the  demonstration  of  usefulness 
can  begin  with  any  of  the  three.  Insulin,  e.g., 
was  first  found  to  have  value  in  therapy; 
fermentation  was  then  shown  to  be  useful  in 
its  production;  and,  now  genetic  engineering 
promises  to  make  the  fermentation  process  eco- 
nomically competitive.  In  contrast,  the  value  of 
thvmosin  a-l,  has  not  vet  been  proved,  although 


56  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


the  usefulness  of  genetic  engineering  and  fer- 
mentation in  its  production  have  been  demon- 
strated. 

As  these  examples  indicate,  the  limits  on  a 
product’s  commercial  potential  vary  with  the 
product.  In  some  cases,  the  usefulness  of  the 
j)i'oduct  has  already  been  shown,  and  the  use- 


fulness of  genetic  technologies  must  be  proved. 
In  others,  the  genetic  technologies  make  pro- 
duction at  the  industrial  level  possible,  hut  their 
market  has  not  yet  been  established.  In  still 
others,  the  feasibility  of  fermentation  is  the  ma- 
jor problem. 


chapter  4 

The  Pharmaceutical  Industry 


chapter  4 


Page 

Background 59 

Past  Uses  of  Genetics 59 

Potential  Uses  of  Molecular  Genetic  Technologies  61 

Hormones 61 

Insulin 65 

Growth  Hormone 67 

Other  Hormones 67 

Immunoproteins 68 

Antigens  (Vaccines) 68 

Interferons 70 

Lymphokines  and  Cytokines 7l 

Antibodies 71 

Enzymes  and  Other  Proteins 72 

Enzymes 72 

Other  Proteins 74 

Antibiotics 75 

Nonprotein  Pharmaceuticals 76 

Impacts 77 

Technical  Notes 80 


Tables 


Table  No.  Page 

3.  Large  Human  Polypeptides  Potentially 

Attractive  for  Biosynthesis 61 


Table  No.  Page 

4.  Naturally  Occurring  Small  Peptides  of 

Potential  Medical  Interest 64 

5.  Summary  of  Potential  Methods  for 

Interferon  Production 70 

6.  Immunoassays 73 

7.  Diseases  Amenable  to  Drugs  Produced  by 
Genetic  Engineering  in  the  Pharmaceutical 

Industry 78 

8.  Major  Diseases  for  Which  Vaccines  Need  To 

Be  Developed 78 


Figures 


. Figure  No.  Page 

20.  The  Development  of  a High  Penicillin- 
Producing  Strain  via  Genetic  Manipulation  . . 60 

21.  The  Product  Development  Process  for 
Genetically  Engineered  Pharmaceuticals  ....  63 

22.  The  Amino  Acid  Sequence  of  Proinsulin  ....  65 

23.  Recombinant  DNA  Strategy  for  Making 

Foot-and-Mouth  Disease  Vaccine 69 


Chapter  4 

The  Pharmaceutical  Industry 


Background 


The  domestic  sales  of  prescription  drugs  by 
L’.S.  pharmaceutical  companies  exceeded  S7.5 
billion  in  1979.  Of  these,  approximately  20  per- 
cent were  products  for  which  fermentation 
processes  played  a significant  role.  They  in- 
cluded anti-infective  agents,  vitamins,  and  bio- 
logicals,  such  as  \ accines  and  hormones,  (ienet- 
ics  is  expected  to  he  particularly  useful  in  the 
production  of  these  pharmaceuticals  and  bio- 
logicals,  which  can  only  be  obtained  by  extrac- 
tion from  human  or  animal  tissues  and  fluids. 

.Although  the  pharmaceutical  industry  was 
the  last  to  adopt  traditional  fermentation  tech- 
nologies, it  has  been  the  first  industry  to  make 
widespread  use  of  such  advanced  genetic  tech- 
nologies as  recombinant  DN'A  (rDN’A)  and  cell 
fusion.  Two  major  factors  triggered  the  use  of 
genetics  in  the  pharmaceutical  industry: 

• The  biological  sources  of  many  pharmaco- 
logically active  products  are  micro-orga- 
nisms, which  are  readily  amenable  to  ge- 
netic engineering. 

• The  major  advances  in  molecular  genetic 
engineering  have  been  made  under  an  in- 
stitutional structure  that  allocates  funds 
largely  to  biomedical  research.  Hence,  the 
Federal  support  system  has  tended  to  fos- 
ter studies  that  hav^e  as  their  ostensible  goal 
the  improvement  of  health. 

Two  factors,  however,  have  tended  to  dis- 
courage the  application  of  genetics  in  the  chem- 
ical and  food  industries.  In  the  former,  econom- 
ic considerations  have  not  allowed  biological 
production  systems  to  be  competitive  with  the 
existing  forms  of  chemical  conv'ersion,  with 
rare  exceptions.  And  in  the  latter,  social  and  in- 
stitutional considerations  hav'e  not  fav'ored  the 
development  of  foods  to  which  genetic  engi- 
neering might  make  a contribution. 


Past  uses  of  genetics 

Genetic  manipulation  of  biological  systems 
for  the  production  of  pharmaceuticals  has  two 
general  goals: 

1.  to  increase  the  lev  el  or  efficiency  of  the 
production  of  pharmaceuticals  with  prov- 
en or  potential  value;  and 

2.  to  produce  totally  new  pharmaceuticals 
and  compounds  not  found  in  nature. 

The  first  goal  has  had  the  strongest  influence 
on  the  industry.  It  has  been  almost  axiomatic 
that  if  a naturally  occurring  organism  can  pro- 
duce a pharmacologically  valuable  substance, 
genetic  manipulation  can  increase  the  output. 
The  following  are  three  classic  examples. 

• The  genetic  improvement  of  penicillin  pro- 
duction is  an  example  of  the  elaborate  long- 
term efforts  that  can  lead  to  dramatic 
increases.  The  original  strains  of  Penicilli- 
um  chrysogenum,  NRRL-1951,  were  treated 
w'ith  chemicals  and  irradiation  through 
successive  stages,  as  shown  in  figure  20, 
until  the  strain  E-15.1  was  developed.  This 
strain  had  a 55-fold  improvement  in  pro- 
ductivity over  the  fungus  in  which  penicil- 
lin was  originally  recognized— the  Fleming 
strain. 

• Chemically  induced  mutations  improved  a 
strain  of  Escherichia  coli  to  the  point  where 
it  produced  over  100  times  more  L-asparag- 
inase  (which  is  used  to  fight  leukemia)  than 
the  original  strain.  This  increase  made  the 
task  of  isolating  and  purifying  the  pharma- 
ceutical much  easier,  and  resulted  in  low- 
ering the  cost  of  a course  of  therapy  from 
nearly  $15,000  to  approximately  $300. 

• Genetic  manipulation  sufficiently  improved 
the  production  of  the  antibiotic,  gentami- 


59 


60  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Figure  20.— The  Development  of  a High  Penicillin- 
Producing  Strain  via  Genetic  Manipulation 


E-15.1  — ► Final  strain 


An  illustration  of  the  extensive  use  of  genetics  to  increase 
the  yield  of  a commercially  valuable  substance.  A variety 
of  laboratories  and  methods  were  responsible  for  the  suc- 
cessful outcome. 

SOURCE:  Adapted  by  Office  of  Technology  Assessment  from  R.  P.  Blander  in 
Genetics  of  Industrial  Microorganisms,  O.  K.  Sebek  and  A.  I.  Laskin 
(eds.)  (Washington,  D.C.:  American  Society  for  Microbiology,  1979), 
p.23. 


cin,  so  that  Schering-Plough,  its  producer, 
did  not  have  to  build  a scheduled  manufac- 
turing plant,  thereby  saving  $50  million. 

Most  industry  analysts  agree  that,  overall, 
genetic  manipulation  has  been  highly  significant 
in  increasing  the  availability  of  many  pharma- 
ceuticals or  in  reducing  their  production  costs. 

The  second  major  goal  of  genetic  manipula- 
tion, the  production  of  new  compounds,  has 
been  achieved  to  a lesser  degree.  A recent  new 
antibiotic,  deoxygentamicin,  was  obtained  by 
mutation  and  will  soon  be  clinically  tested  in 
man.  Earlier,  an  important  new  antibiotic, 
amikacin,  was  produced  through  classical  mo- 
lecular genetic  techniques.  And  before  that,  the 
well-known  antibiotic,  tetracycline,  which  is 
normally  not  found  in  nature,  was  produced  by 
a strain  of  the  bacterium,  Streptomyces,  after 
appropriate  genetic  changes  had  been  carried 
out  in  that  bacterium. 


Ch.  4— The  Pharmaceutical  Industry  • 61 


Potential  uses  of  molecular  genetic  technologies 


l\)lypeptities— proteins— are  the  tirst  abun- 
dant end  prodiiets  of  genes.  Thev  inelude  pep- 
tide hormones,  enzymes,  antibodies,  and  cer- 
tain \aeeines.  Producing  tliem  is  the  goal  of 
most  current  efforts  to  harness  genetically 
directed  processes.  Houe\er,  it  is  just  a matter 
of  time  and  the  exolution  of  technolog\'  before 
complex  non[)roteins  like  antibiotics  can  also  he 
manufactured  through  rI)>J,\  techni(|ues. 

Hormones 

The  most  ad\anced  apf)lications  of  genetics 
today,  in  terms  of  technological  sophistication 
and  commercial  de\  elopment,  are  in  the  field  of 
hormones,  the  potent  messenger  molecules  that 
help  the  body  cooi  dinate  the  actions  of  \ arious 
tissues.  (See  Tech.  Note  1,  p.  80.)  The  capacity  to 
synthesize  proteins  through  genetic  engineer- 
ing has  stemmed  in  large  part  from  attempts  to 
prepare  human  peptide  hormones  (like  insulin 
and  growth  hormone).  The  diseases  caused  by 
their  deficiencies  are  presently  treated  with  ex- 
tracts made  from  animal  or  human  glands. 

The  merits  of  engineering  other  peptide  hor- 
mones depend  on  understanding  their  actions 
and  those  of  their  deri\ati\es  and  analogs. 
E\idence  that  they  might  be  used  to  improxe 
the  treatment  of  diabetes,  to  promote  wound 
healing,  or  to  stimulate  the  regrowth  of  nerv'es 
will  stimulate  new  scientific  investigations. 
Other  relati\ely  small  polypeptides  that  influ- 
ence the  sensation  of  pain,  appetite  suppression, 
and  cognition  and  memory  enhancement  are 
also  being  tested.  If  they  prove  useful,  they  will 
unquestionably  be  evaluated  for  production  via 
fermentation. 

VV'hile  certain  hormones  have  already  at- 
tained a place  in  pharmacology,  their  testing 
and  use  has  been  hindered  to  some  extent  by 
tbeir  scarcity  and  high  cost.  Until  recently, 
animal  glands,  human-cadaver  glands,  and 
urine  were  the  only  sources  from  which  they 
could  be  drawn.  Their  use  is  also  limited 
because  polypeptide  hormones  must  be  ad- 
ministered bv  injection.  Thev  are  digested  if 


they  are  taken  orally,  a [)rocess  that  curtails 
their  usefulness  and  causes  side-effects. 

Thei'e  are  four  technologies  for  producing 
[)oly peptide  hormones  and  polypeptides: 

• extraction  from  human  or  animal  organs, 
sei'um,  or  urine; 

• chemical  synthesis; 

• |)i'oduction  by  cells  in  tissue  culture;  and 

• production  by  microbial  fermentation  after 
genetic  engineering. 

One  major  factor  in  deciding  which  technol- 
ogy is  best  for  which  hormone  is  the  length  of 
the  hormone’s  amino  acid  chains.  (See  table  3.) 
Modern  methods  of  chemical  synthesis  have 
made  the  preparation  of  low-molecular  weight 
polypeptides  a fairly  straightforward  task,  and 
chemically  synthesized  hormones  up  to  at  least 
32  amino  acids  (AA)  in  length— like  calcitonin 


Table  3.— Large  Human  Polypeptides  Potentially 
Attractive  for  Biosynthesis 


Amino  acid 
residues 

Molecular 

weight 

Prolactin 

. . . 198 

Placental  lactogen 

. . . 192 

'Growth  hormone 

. ..  191 

22,005 

Nerve  growth  factor 

...  118 

13,000 

Parathyroid  hormone  (PTH) . . . 

. . . 84 

9,562  bovine 

Proinsulin 

. . . 82 

Insulin-like  growth  factors 
(IGF-I  &IGF-2) 

. . . 70,  67 

7,649,  7471 

Epidermal  growth  factor 

6,100 

'Insulin 

. . . 51 

5,734 

Thymopoietin 

. . . 49 

Gastric  inhibitory  polypeptide 
(GIP) 

. . . 43 

5,104  porcine 

'Corticotropin  (ACTH) 

. . . 39 

4,567  porcine 

Cholecystokinin  (CCK-39)  . . . . 

. . . 39 

Big  gastrin  (BG) 

. . . 34 

Active  fragment  of  PTH 

. . . 34 

4,109  bovine 

Cholecystokinin  (CCK-33)  . . . . 

. . . 33 

3,918  porcine 

'Calcitonin 

. . . 32 

3,421  human 

Endorphins 

. . . 31 

3,435  salmon 
3,465 

'Glucagon 

. . . 29 

3,483  porcine 

Thymosin-<yt 

. . . 28 

3,108 

Vasoactive  intestinal  peptide  (VIP)  28 

3,326  porcine 

'Secretin 

. . . 27 

'Active  fragment  of  ACTH .... 

. . . 24 

Motilin 

. . . 22 

2,698 

'Currently  used  in  medical  practice. 
SOURCE:  Office  of  Technology  Assessment. 


62  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


—have  become  competitive  with  those  derived 
from  current  biological  sources.  Since  frag- 
ments of  peptide  hormones  often  express  activ- 
ities comparable  or  sometimes  superior  to  the 
intact  hormone,  a significant  advantage  of 
chemical  synthesis  for  research  purposes  is  that 
analogs  having  slight  pharmacological  differ- 
ences from  natural  hormones  can  be  prepared 
by  incorporating  different  amino  acids  into 
their  structures.  In  principle  however,  geneti- 
cally engineered  biosynthetic  schemes  can  be 
devised  for  most  desirable  peptide  hormones 
and  their  analogs,  although  the  practicality  of 
doing  so  must  be  assessed  on  a case-by-case 
basis.  Ultimately,  tbe  principal  factors  bearing 
on  the  practicality  of  the  competing  alternatives 
are: 

• The  cost  of  raw  materials.  For  genetically 
engineered  biosynthesis,  this  includes  the 
cost  of  the  nutrient  broth  plus  some  amor- 
tization of  the  cost  of  developing  the  syn- 
thetic organism.  In  the  case  of  chemical 
synthesis,  it  includes  the  cost  of  the  pure 
amino  acid  subunits  plus  the  chemicals 
used  as  activating,  protecting,  coupling,  lib- 
erating, and  supporting  agents  in  the  proc- 
ess. 

• The  different  costs  of  separating  the  de- 
sired product  from  the  cellular  debris  and 
tbe  culture  medium  in  biological  produc- 
tion, and  from  tbe  supporting  resin,  by- 
products, and  excess  reagents  in  chemical 
synthesis. 

• The  cost  of  purification  and  freedom  from 
toxic  contaminants.  The  process  is  more 
expensive  for  biologically  produced  materi- 
al than  for  materials  produced  by  conven- 
tional chemistry,  although  hormones  from 
any  source  can  be  contaminated. 

• Differences  in  the  costs  of  labor  and  equip- 
ment. Chemical  synthesis  involves  a se- 
quence of  similar  (but  different)  operations 
during  a time  period  roughly  proportional 
to  the  length  of  the  amino  acid  chain  (three 
AA  per  day)  in  an  apparatus  large  enough 
to  produce  100  grams  (g)  to  1 kilogram  (kg) 
per  batch;  biological  fermentations  use  vats 
—with  capacities  of  several  thousand  gal- 
lons—for  a few  days,  regardless  of  the 
length  of  the  amino  acid  chain. 


• The  cost  and  suitability  of  comparable 
materials  gathered  from  organs  or  fluids 
obtained  from  animals  or  people. 

In  the  past  decade,  some  simpler  hormones 
have  been  chemically  synthesized  and  a few  are 
being  marketed.  However,  synthesizing  glyco- 
proteins—proteins  bound  to  carbohydrates— is 
still  beyond  the  capabilities  of  chemists.  Data 
obtained  from  companies  directly  inxolved  in 
the  production  of  peptides  by  chemical  synthe- 
sis indicate  that  the  cost  of  chemically  preparing 
polypeptides  of  up  to  50  AA  in  length  is  ex- 
tremely sensitive  to  volume  (see  Tech.  Note  2,  p. 
80.);  although  the  costs  are  high,  the  production 
of  large  quantities  by  chemical  synthesis  offers 
a competitive  production  method. 

Nevertheless,  rDNA  production,  also  known 
as  molecular  cloning,  has  already  been  used  to 
produce  low-molecular  weight  polypeptides.  In 
1977,  researchers  at  Genentech,  Inc.,  a small 
biotechnology  company  in  California,  inserted  a 
totally  synthetic  DNA  sequence  into  an  E.  coli 
plasmid  and  demonstrated  that  it  led  to  the  pro- 
duction of  the  14  AA  polypeptide  seciuence  cor- 
responding to  somatostatin,  a hoi'inone  found  in 
the  brain.  The  knowledge  of  somatostatin’s 
amino  acid  sequence  made  the  experiment  pt)s- 
sible,  and  the  existence  of  sensiti\e  assays  al- 
lowed the  hormone’s  expression  to  be  detect('d. 
Although  the  primary  motive  foi’  using  this  par- 
ticular hormone  for  the  first  demonstration  was 
simply  to  show  that  it  could  he  cIoik?,  (ien(Mit('ch 
has  announced  that  it  plans  to  mark(>t  its 
genetically  engineered  molecule  foi’  r(\s(^ar('h 
purposes.  (See  figure  21.) 

Somatostatin  is  one  of  about  20  i'ecogniz('d 
small  human  polypeptides  that  can  he  made 
without  difficulty  hy  chemical  synthesis.  (Se(* 
table  4.)  Unless  a sizable  market  is  found  foi"  one 
of  them,  it  is  unlikely  that  fei  inentation  meth- 
ods will  be  developed  in  tbe  foreseeable  luture. 
Some  small  peptides  that  may  justify  tlie  dewl- 
opment  of  a biosynthetic  process  of  production 
are: 

• The  seven  AA  seciuenci*  known  as  MSN 

ACTH  4-10,  w'hich  is  reputed  to  influence 
memory,  concentration,  and  other  p.sycho- 
logical-hehavioral  ('fleets:  should  such 


Ch.  4— The  Pharmaceutical  Industry  • 63 


Figure  21.— The  Product  Development  Process  for  Genetically  Engineered  Pharmaceuticals 

Micro-organisms  such  as  E.  coll 

© «) 


19.  Submit  IND 


The  development  process  begins  by  obtaining  DNA  either  through  organic  synthesis  (1)  or  derived  from  biological  sources  such  as  tissues 
(2).  The  DNA  obtained  from  one  or  both  sources  is  tailored  to  form  the  basic  “gene"  (3)  which  contains  the  genetic  information  to  “code"  for  a 
desired  product,  such  as  human  interferon  or  human  insulin.  Control  signals  (4)  containing  plasmids  (6)  are  isolated  from  micro-organisms  such 
as  E.  coir,  cut  open  (7)  and  spliced  back  (8)  together  with  genes  and  control  signals  to  form  "recombinant  DNA"  molecules.  These  molecules  are 
then  introduced  into  a host  cell  (9). 

Each  plasmid  is  copied  many  times  in  a cell  (10).  Each  cell  then  translates  the  information  contained  in  these  plasmids  into  the  desired  pro- 
duct. a process  called  "expression"  (11).  Cells  divide  (12)  and  pass  on  to  their  offspring  the  same  genetic  information  contained  in  the  parent 
cell. 

Fermentation  of  large  populations  of  genetically  engineered  micro-organisms  is  first  done  in  shaker  flasks  (13),  and  then  in  small  fermenters 
(14)  to  determine  growth  conditions,  and  eventually  in  larger  fermentation  tanks  (15).  Cellular  extract  obtained  from  the  fermentation  process  is 
then  separated,  purified  (16),  and  packaged  (17)  for  health  care  applications. 

Health  care  products  are  first  tested  in  animal  studies  (18)  to  demonstrate  a product’s  pharmacological  activity  and  safety.  In  the  United 
States,  an  investigational  new  drug  application  (19)  is  submitted  to  begin  human  clinical  trials  to  establish  safety  and  efficacy.  Following 
clinical  testing  (20),  a new  drug  application  (NDA)  (21)  is  filed  with  the  Food  and  Drug  Administration  (FDA).  When  the  NDA  has  been  reviewed 
and  approved  by  the  FDA  the  product  may  be  marketed  in  the  United  States  (22). 


SOURCE;  Genentech.  Inc. 


64  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


Table  4.— Naturally  Occurring  Small  Peptides  of 
Potential  Medical  Interest 


Number  of 
amino  acids 

Molecular 

weight 

Dynorphin 

17 

Little  gastrin  (LG) 

17 

2,178 

Somatostatin 

14 

1,639 

Bombesin 

14 

1,620 

Melanocyte  stimulating  hormone. 

13 

1,655 

Active  dynorphin  fragment 

13 

Neurotensin 

13 

Mini-gastrin  (G13) 

13 

Substance?  

Luteinizing  hormone-releasing 

11 

1,347  bovine 

hormone  (LNRH) 

10 

1,183 

Active  fragment  of  CCK 

10 

Angiotensin  1 

10 

1,297 

Caerulein 

10 

1,252  porcine 

Bradykinin 

9 

1,060 

'Vasopressin  (ADH) 

9 

'Oxytocin 

9 

1,007 

Facteur  thymique  serique  (FTH) . . 

9 

Substance  P(4-11)octapeptide. . . 

8 

966 

Angiotensin  II 

8 

1,046 

Angiotensin  III 

7 

931 

MSH/ACTH4-10 

7 

Enkephalins 

Active  fragment  of  thymopoietin 

5 

575 

(TP5) 

'Thyrotropin  releasing  hormone 

5 

(TRH) 

3 

362 

•Currently  used  in  medical  practice. 
SOURCE:  Office  of  Technology  Assessment. 


agents  prove  of  value  in  wider  testing,  they 
have  an  enormous  potential  for  use. 

• Both  cholecystokinin  (33  AA)  and  bomhesin 
(10  AA),  which  have  been  shown  to  sup- 
press appetite,  presumably  as  a satiety 
signal  from  stomach  to  brain:  there  is  a 
large  market  for  antiobesity  agents— ap- 
proximately $85  million  per  year  at  the 
manufacturer’s  level. 

• Several  hormones,  such  as  somatostatin, 
which  are  released  by  nerves  in  the  hypo- 
thalamus of  the  brain  to  stimulate  or  in- 
hibit release  of  hormones  by  the  pituitary 
gland:  hormones  produced  by  these  glands 
are  crucial  in  human  fertility;  analogs  of 
some  are  being  investigated  as  possible 
contraceptives. 

• Calcitonin  (32  AA),  which  is  currently  the 
largest  polypeptide  produced  by  chemical 
synthesis  for  commercial  pharmaceutical 
use:  it  is  useful  for  pathologic  bone  dis- 
orders, such  as  Paget’s  disease,  that  affect 


up  to  3 percent  of  the  population  over  40 
years  of  age,  in  Western  Europe. 

• Adrenocorticotropic  hormone  (ACTH)  (39 
AA),  which  promotes  and  maintains  the 
normal  growth  and  development  of  the 
adrenal  glands  and  stimulates  the  secretion 
of  other  hormones:  in  the  United  States, 
ACTH  is  used  primarily  as  a diagnostic 
agent  for  adrenal  insufficiency,  but  in 
principle,  ACTH  might  be  used  for  at  least 
one-third  of  the  medical  indications— like 
rheumatic  disorders,  allergic  states,  and 
eye  inflammation— for  which  about  5 mil- 
lion Americans  annually  recei\e  corticos- 
teroids. 

Within  the  last  5 years,  other  small  polypep- 
tides have  been  identified  in  many  tissues  and 
have  been  linked  to  a \arietv  of  activ  ities.  Some 
certainly  bind  to  the  same  receptor  sites  as  the 
pain-relieving  opiates  related  to  the  morphine 
family.  These  peptides  are  called  endogenous 
opiates:  the  smaller  (5  AA)  peptides  are  called 
enkephalins  and  the  larger  (3 1 AA),  endoi'phins. 

Certain  enkephalins  produce  hi'ief  analgesia 
when  injected  directly  into  the  hi’ains  of  mice. 
Synthetic  analogs  that  are  less  susceptible  to  en- 
zymatic inactivation  produce  longer  analgesia 
even  if  they  are  injected  intravenously,  as  does 
the  larger  j8-endorphin  molecule.  \'(M’v  reccMilly, 
a 17  AA  polypeptide,  dynoi’phin,  was  r(>ported 
to  be  the  most  potent  pain  killer  yet  found— it  is 
1,200  times  more  powerful  than  morphine. 

The  preparation  of  new  analgesic  agents  ap- 
pears a likely  outcome  of  the  ncnv  research,  hut 
problems  similar  to  those  associated  with  clas- 
sical opiates  must  he  overcome.  (T)nse(|uentl\ , 
unnatural  analogs— including  some  made  with 
amino  acids  not  found  in  mici'o-organisms— 
might  prove  more  useful.  The  value  of  microhi- 
al  biosynthesis  for  these  substances  is  (jiies- 
tionable  at  this  time.  Howcvcm',  the  im|)ortance 
of  genetic  technologies  in  clarilving  the 
underlying  mechanisms  should  tiot  he  undei - 
estimated. 

Higher  moleculai'  weight  [)olv[)eptldes  cannot 
be  made  practically  by  chemical  synthesis,  .ind 
must  he  exti’acted  from  human  or  animal  tis 
sues  or  produced  in  cells  growing  in  culture 


Ch.4 — The  Pharmaceutical  Industry  • 65 


\ow  they  can  also  be  mamit'actured  by  fermen- 
tation using  genetically  designed  bacteria,  as 
has  been  demonstrated  by  the  production  of  in- 
sulin and  human  grow  tb  hormone. 

I\SI  Ll\ 

Insulin,  is  composed  of  tw  o chains— -\  and  B— 
of  amino  acids.  It  is  initially  produced  as  a 
single,  long  chain  called  pre-|)i'oinsulin,  which  is 
cut  into  a shorter  chain,  proinsulin.  Proinsulin, 
in  turn,  is  cut  into  the  ,\  anti  B chains  w hen  a 
piece  is  cleav  ed  from  the  middle.  (See  figure  22.) 
\\  ork  on  the  genetic  engineering  of  insulin  has 


pi'oceeded  quickly.  ,-\  year  after  one  group  re- 
ported that  the  insulin  gene  had  been  incorpo- 
I’ated  into  E.  coli  without  e.xpression,  a second 
group  managed  to  grow  colonies  of  £.  coli  that 
actually  e.xcreted  rat  [)roinsulin.  Then,  within  a 
couple  of  months,  workers  at  (Jenentech,  in  col- 
laboration with  a grouf)  at  City  of  Hope  Medical 
Center,  announced  the  se|)arate  synthesis  of  the 
.\  (2  1 .\,-\)  anti  B (30  .A, A)  chains  of  human  insulin. 
The  synthesis  of  the  1)N,A  secjuences  depended 
on  advances  in  organic  chemisti’v  as  well  as  in 
genetics.  Six  months  were  required  simply  to 
synthesize  the  necessary  building  blocks. 


Figure  22.— The  Amino  Acid  Sequence  of  Proinsulin 


Connecting  peptide 


A chain 


B chain 

Proinsulin  is  composed  of  84  amino  acid  residues.  When  the  connecting  peptide  is  removed, 
the  remaining  A and  B chains  form  the  insulin  molecule.  The  A chain  contains  21  amino  acids; 
the  B chain  contains  30  amino  acids. 


SOURCE:  Office  of  Technology  Assessment. 


66  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


A comparison  with  the  traditional  source  of 
animal  insulin  is  interesting.  If  0.5  milligram 
(mg)  of  pure  insulin  can  be  obtained  from  a liter 
of  fermentation  brew,  2,000  liters  (1)  (roughly 
500  gal)  would  yield  1 g of  purified  insulin— the 
amount  produced  by  about  16  lb  of  animal  pan- 
creas. If,  on  the  other  hand,  the  efficiency  of 
production  could  be  increased  to  that  achieved 
for  asparaginase  (which  is  produced  commer- 
cially by  the  same  organism,  E.  coli),  2,000  1 
would  yield  100  g of  purified  insulin— the 
amount  extracted  from  1,600  lb  of  pancreas. 
(The  average  diabetic  uses  the  equivalent  of 
about  2 mg  of  animal  insulin  per  day.) 

The  extent  of  the  actual  demand  for  insulin  is 
a controversial  issue.  Eli  Lilly  & Co.  estimates 
that  there  are  60  million  diabetics  in  the  world 
(35  million  in  underdeveloped  countries,  where 
few  are  diagnosed  or  treated).  Of  the  25  million 
in  the  developed  countries,  perhaps  15  million 
have  been  diagnosed;  according  to  Lilly’s  esti- 
mate, 5 million  are  treated  with  insulin.  Only 
one-fourth  of  those  diabetics  treated  with  in- 
sulin live  in  the  United  States,  but  they  use  40  to 
50  percent  of  the  insulin  consumed  in  the 
world.  A number  of  studies  indicate  that  while 
the  emphasis  on  diet  (alone)  and  oral  antidia- 
betic drugs  varies,  approximately  40  percent  of 
American  patients  in  large  diabetes  clinics  or 
practices  take  insulin  injections.  In  the  United 
States,  diabetes  ranks  as  the  fifth  most  common 
cause  of  death  and  second  most  common  cause 
of  blindness.  Roughly  2 million  persons  require 
daily  injections  of  insulin. 

Today,  at  least,  there  is  no  real  shortage  of 
glands  from  slaughter  houses  for  the  produc- 
tion of  animal  (principally  bovine  and  porcine) 
insulin.  A study  conducted  by  the  National  Dia- 
betes Advisory  Board  (NDAB)  concluded  that  a 
maximum  demand  and  a minimum  supply 
would  lead  to  shortages  in  the  1990’s.  Eli  Lilly’s 
projection,  presented  in  that  report,  also  antici- 
pates these  shortages.  But,  Novo  Industri,  a ma- 
jor world  supplier  of  insulin,  told  the  NDAB  that 
it  estimates  that  the  1976  free-world  consump- 
tion of  insulin  of  51  X 10®  units  constituted  only 
23  percent  of  the  potential  supply,  and  the 
87X10®  units  projected  for  1996  would  only 
equal  40  percent  of  the  supply,  assuming  that 
the  animal  population  stays  constant. 


Lor  insulin,  therefore,  the  limitation  on  bring- 
ing the  fruits  of  genetic  engineering  to  the 
marketplace  is  not  technological  hut  institu- 
tional. The  drug  must  first  be  appro\-ed  by  the 
Eood  and  Drug  Administration  (FD.A)  and  then 
marketed  as  a product  as  good  as  or  better  than 
the  insulin  extracted  by  con\entional  means. 
Lilly  has  stated  that  it  anticipates  a 6-month 
testing  period  in  humans.  Undoubtedly,  LD.A 
will  examine  the  e\  idence  presented  in  the  in- 
vestigational new  drug  a[)|)lication  (IND.A)  w ith 
special  care.  Its  rexiew  will  establish  criteria 
that  may  influence  the  rex  iew  of  suhs(‘(|uent  aj)- 
plications  in  at  least  the  folloxx  ing  re(|uirements: 

• evidence  that  the  amino  aciti  .se(|uence  of 
the  material  is  identical  to  that  of  the  nor- 
mal human  hormone: 

• freedom  from  hactei'ial  endoto.xins  that 
may  cause  lex  er  at  exti'cim'lx  loxx  concen- 
trations—an  inhei’ent  hazard  as.sociated 
xvith  any  process  using  E.  coli;  and 

• freedom  from  byproducts,  including  sub- 
stances of  xei'v  similar  structure  that  max’ 
give  rise  to  rare  a('ut(>  or  chronic  r»*actions 
of  the  immune  system. 

Furthermore,  as  dex’elopment  continues,  IDA 
might  recjuire  strict  assurances  that  tlu*  mole- 
cules j)i'oduced  from  hatch  to  hatch  are  not  sub- 
ject to  subtle  xai’iations  resulting  Irom  their 
genetic  origin. 

If  the  insulin  obtained  from  rD.VA  techniques 
manages  to  pass  1D,\  i'e(|uirements,  it  must 
oxei'come  a second  obstacle— competition  in  the 
marketplace.  I’he  clinical  rationale  lor  using 
human  rather  than  animal  insulin  rests  on  the 
differences  in  structure  among  insulins  pro- 
duced by  different  species  Human  .md  porcine 
insulins  for  example,  differ  in  a single  amino 
acid,  xvhile  human  and  cattle  insulins  diller 
xvith  respiH't  to  three.  .As  lar  .is  is  known  these 
variations  do  not  impair  the  elleclixenes  o|  the 
insulin,  hut  no  om*  has  i‘x  (*r  been  in  a |M)sition  to 
conduct  a significant  lest  ol  the  use  ot  hum.m 
insulin  in  a dialxMic  |)0|)ulalion  M.inx  lonse 
(luences  of  thi;  dise.ise,  such  as  ri'linopatlix  tret 
inal  diseas(')  and  n(>phro|)athx  Ikidnex  dise.isel 
are  not  prm’ented  by  routine  injection  of  .mim.il 
insulin.  Lati(‘nts  also  occasionallx  respond  .id 
versely  or  produce  antibodies  to  .inim.il  insulin 
xvith  suhs('(|uenl  allergic  or  resist. ml  re. k lion 


Ch.4 — The  Pharmaceutical  Industry  • 67 


It  remains  to  be  seen  liow  many  patients  will 
ibe  bettei’ oft  \\  itb  human  insulin.  The  |)root  that 
lit  improv  es  tlu*ra[n  w ill  take  years.  F’rogress  on 
|the  etiologv  ot  the  ilisease— espeeially  in  itlenti- 
living  it  in  those  at  risk  or  in  improving  th(> 
iiiosage  form  and  administration  of  insulin— may 
have  far  more  significant  effects  than  new  de- 
\el()[)ments  in  insulin  {)roduction.  Nevertheless, 
as  long  as  priv  ate  enterprise  sc?es  fit  to  inv fst  in 
|such  develo[)ments,  and  as  long  as  the  cost  of 
{treating  diabetics  w ho  rtvspond  pi’operly  to  ani- 
linal  insulin  is  not  inci'eased.  biological  produc- 
'tion  of  human  insulin  mav  hcH  ome  a kind  of  in- 
Isurance  for  diahc'tics  within  the  next  few 
idcH'ades. 

ciHow  I II  iicmMovt: 

The  second  polvpc*ptide  hormone  currently  a 
(candidate  for  KI).\  apfirov  al  is  gi'ow  th  hormone 
|(CiHI.  It  is  one  of  a family  of  closely  l elated,  rel- 
tatively  large  pituitary  peptide  hormones— sin- 
gle-chain polypeptides  191-  to  19«-,\.\  in  length. 
It  is  best  known  for  the  growth  it  induces  in 
I many  soft  tissues,  cartilage,  and  hone,  and  it  is  a 
Ireciuirement  for  jxistnatal  grow  th  in  man. 

^ rhe  grow  th  of  an  organism  is  a highlv  com- 
jple.x  process  that  depends  on  the  correct  hal- 
lance  of  many  variables:  I he  action  of  C'lH  in  the 
I body  for  example,  depends  on  the  presence  of 
j insulin,  whose  secretion  is  stimulated  by  GH. 

! Under  some  circumstances,  one  or  more  inter- 
Imediarv  polypeptides  produced  under  the  in- 
[tluence  of  GH  by  the  liver  (and  possibly  the 
I kidneys)  may  actually  be  the  proximate  causes 
I of  some  of  the  effects  attributed  to  GH.  In  any 
'case,  the  biological  significance  of  GH  is  most 
'clearly  illustrated  by  the  growth  retardation 
> that  characterizes  its  absence  before  puberty, 

! and  bv  the  benefits  of  replacement  therapy. 

In  the  United  States,  most  of  the  demand  for 
I human  growth  hormone  (hGH)  is  met  by  the  Na- 
tional Pituitary  Agency,  which  was  created  in 
I the  early  1960’s  by  the  College  of  Pathologists 
I and  the  National  Institute  of  Arthritis,  Metab- 
: olism,  and  Digestive  Diseases  (NIAMDD)  to  col- 
} lect  pituitary  glands  from  coroners  and  private 
I donors.  Under  the  programs  of  the  NIAMDD, 

I hGH  is  prov  ided  without  charge  to  treat  chil- 
' dren  with  hypopituitarism,  or  dwarfism  (about 
( 
t 
i 


l.tiOO  patients,  each  of  whom  receives  therapy 
for  several  years),  and  for  research. 

While  the  National  Pituitary  Agency  feels  that 
it  can  satisfy  the  current  demand  for  hCiH  (see 
Tech.  Note  3,  p.  80.),  it  welcomes  the  promise  of 
ailditional  hGH  at  relatively  low  cost  to  satisfy 
ai'eas  of  research  that  are  handicapped  more  by 
a scarcity  of  funds  than  by  a scarcity  of  the  hor- 
mone. However,  if  hGH  is  shown  to  he  thera- 
peutically valuable  in  these  areas,  widespread 
use  could  severely  strain  the  present  supply.  At 
present,  the  potential  seems  greatest  for  pa- 
tients with: 

• senile  osteo[)orosis  (hone  decalcification); 

• other  nonpituitary  growth  deficiences  such 
as  lurner’s  syndi'ome  (1  in  3,000  live 
female  births); 

• intrauterine  growth  retardation; 

• bleeding  ulcers  that  cannot  be  controlled 
by  other  means;  and 

• burn,  wound,  and  hone-fracture  healing 

Two  groups  have  already  announced  the 
preparation  of  micro-organisms  with  the  capaci- 
ty for  synthesizing  GH.  (See  Tech.  Note  4,  p.  80.) 
In  December  1979,  one  of  these  groups— Genen- 
tech  —requested  and  received  permission  from 
the  National  Institutes  of  Health  (NIH),  on  the 
recommendation  of  the  Recombinant  DNA  Ad- 
visory Committee  (RAC),  to  scale-up  its  process. 
Its  formation  of  a joint-venture  with  Kabi  Gen 
,AB  is  typical  of  the  kind  of  alliance  that  develops 
as  a result  of  the  different  expertise  of  groups  in 
the  multidisciplinary  biomedical  field.  Kabi  has 
been  granted  a New  Drug  Application  (NDA) 
under  which  to  market  pituitary  GH  imported 
from  abroad. 

OTHER  HORMONES 

Additional  polypeptide  hormones  targeted 
for  molecular  cloning  (rDNA  production)  in- 
clude: 

• Parathyroid  hormone  (84  AA),  which  may 
be  useful  alone  or  in  combination  with  cal- 
citonin for  bone  disorders  such  as  osteo- 
porosis. 

• Nerve  growth  factor  (118  AA),  which  influ- 
ences the  development,  maintenance,  and 


68  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


repair  of  nerve  cells  and  thus  could  be  sig- 
nificant for  nerve  restoration  in  surgery. 

• Erythropoietin,  a glycopeptide  that  is  large- 
ly responsible  for  the  regulation  of  blood 
cell  development.  Its  therapeutic  applica- 
tions may  range  from  hemorrhages  and 
burns  to  anemias  and  other  hematologic 
conditions.  (See  Tech.  Note  5,  p.  80.) 

Immunoproteins 

Immunoproteins  include  all  the  proteins  that 
are  part  of  the  immune  system— antigens,  inter- 
ferons, cytokines,  and  antibodies.  Since  poly- 
peptides, the  primary  products  of  every  molec- 
ular cloning  scheme,  are  at  the  heart  of  immu- 
nology, developments  made  possible  by  recent 
breakthroughs  will  presumably  affect  the  entire 
field.  There  is  little  doubt  that  applied  genetics 
will  play  a critical  role  in  developing  a pharma- 
cology for  controlling  immunologic  functions, 
since  it  provides  the  only  apparent  means  of 
synthesizing  many  of  the  agents  that  will  com- 
prise immunopharmacology. 

ANTIGENS  (VACCINES) 

One  early  dramatic  benefit  should  be  in  the 
area  of  vaccination,  where  genetic  technologies 
may  lead  to  the  production  of  harmless  sub- 
stances capable  of  eliciting  specific  defenses 
against  various  stubborn  infectious  diseases. 

Vaccination  provides  effective  immunity  by 
introducing  relatively  harmless  antigens  into 
the  immune  system  thereby  allowing  the  body 
to  establish,  in  advance,  adequate  levels  of  anti- 
body and  a primed  population  of  cells  that  can 
grow  when  the  antigen  reappears  in  its  virulent 
form.  Obviously,  however,  the  vaccination  itself 
should  not  be  dangerous.  As  a result,  several 
methods  have  been  developed  over  the  past  two 
centuries  to  modify  the  virulence  of  micro-orga- 
nisms used  in  vaccines  without  destroying  their 
ability  to  trigger  the  production  of  antibodies. 
(See  Tech.  Note  6,  p.  80.) 

Novel  pure  vaccines  based  on  antigens  syn- 
thesized by  rDNA  have  been  proposed  to  fight 
communicable  diseases  like  malaria,  which  have 
resisted  classical  preventive  efforts.  Pure  vac- 
cines have  always  been  scarce;  if  they  were 
available,  they  might  reduce  the  adverse  effects 


of  conventional  vaccines  and  change  the  meth- 
ods and  the  dosages  in  which  vaccines  are 
administered. 

Some  vaccines  are  directed  against  toxic  pro- 
teins (like  the  diphtheria  toxin  produced  by 
some  organisms),  preparing  the  body  to  neutral- 
ize them.  Molecular  cloning  might  make  it  pos- 
sible to  produce  inactivated  toxins,  or  better 
nonvirulent  fragments  of  toxins,  by  means  of 
micro-organisms  that  are  incapable  of  seiz  ing 
as  disease-causing  organisms. 

Immunity  conferred  by  live  vaccines  invari- 
ably exceeds  that  conferred  by  nonli\  ing  anti- 
genic material— possibly  because  a living  micro- 
organism creates  more  antigen  over  a longer 
period  of  time,  providing  continuous  "booster 
shots.”  Engineered  micro-organisms  might  be- 
come productive  sources  of  high-potency  anti- 
gen, offering  far  larger,  more  sustained,  doses 
of  vaccine  without  the  side-effects  from  the  con- 
taminants found  in  those  vaccines  that  consist 
of  killed  micro-organisms. 

However,  it  is  clear  that  formidable  Federal 
regulatory  requirements  would  ha\  e to  he  met 
before  permission  is  granted  for  a no\  el  li\  ing 
organism  to  he  injected  into  human  subjects. 
Because  of  problems  encountered  with  li\  e \ ac- 
cines,  the  most  likely  application  will  lie  in  the 
area  of  killed  vaccines  (often  using  only  parts  of 
micro-organisms). 

It  is  impossible  in  the  scope  of  this  r('port  to 
discuss  the  pros,  cons,  and  conse(|U(‘nc(?s  of  de- 
veloping a vaccine  for  each  viral  disea.se.  How- 
ever, the  most  commercially  important  are  tin* 
influenza  vaccines,  with  an  a\'(M’age  of  20.8  mil- 
lion doses  given  per  year  from  1973  to  197.'>— a 
smaller  number  than  the  25.0  million  doses  |)er 
year  of  polio  vaccine,  hut  moix^  profitable. 

Influenza  is  caused  by  a \ ii'us  that  has  re- 
mained unconti'oll(;d  larg(;ly  because  of  the  fre- 
quency with  which  it  cati  mutate  and  change  its 
antigenic  structures.  It  has  h(‘(Mi  suggested  that 
antigenic  protein  genes  for  influenza  could  he 
kept  in  a "gene  hank”  and  used  w hen  nec'ded  Iti 
addition,  the  genetic  code  for  several  antigens 
could  he  introduced  into  an  organism  such  as  / 


Ch.4 — The  Pharmaceutical  Industry  • 69 


coli,  so  that  a \accine  witli  se\eral  antigens 
might  he  produced  in  one  fei'inentation.' 

Tu  o more  \ iral  diseases  deser\  e at  least  brief 
comment.  .Appro.ximately  800  million  doses  of 
foot-and-mouth  disease  \ irus  (FMD\  ) \ accine 
are  annually  used  worldwide,  making  it  the 
largest  \ olume  \ accine  produced.  This  vaccine 
must  be  given  frequently  to  livestock  in  areas 
where  the  disease  is  endemic,  which  includes 
most  of  the  world  outside  of  North  .America. 
The  present  methods  of  producing  the  vaccine 
require  that  enormous  (juantities  of  hazardous 
\ irus  he  contained.  Many  outbreaks  are  attrib- 
uted to  incompletely  inactivated  vaccine  or  to 
the  escape  of  the  virus  from  factories.  (See 
figure  23.) 

Molecular  cloning  of  the  antigen  could  pro- 
duce a stable  vaccine  at  considerably  less  ex- 
pense, vv  ithout  the  risk  of  the  virus  escaping.  On 
the  basis  of  that  potential,  R.AC  has  approved  a 
joint  program  between  the  U.S.  Department  of 
Agriculture  (L'SDA)  and  Genentech  to  clone 
pieces  of  the  FMD\’  genome  to  produce  pure  an- 
tigen. The  RAC  decision  marked  the  first  excep- 
tion to  the  N'lFl  prohibition  against  cloning  DNA 
that  is  derived  from  a virulent  pathogen.^  FMDV 
vaccine  made  by  molecular  cloning  will  prob- 
ably be  distributed  commercially  by  1985,  al- 
though not  in  the  United  States.  It  will  be  the 
first  vaccine  to  achieve  that  status,  and  illus- 
trates the  potential  veterinary  uses  of  genetic 
technologies. 

Hepatitis  has  also  received  significant  atten- 
tion. Vaccines  against  viral  hepatitis,  which  af- 
fects some  300,000  Americans  each  year,  may 
be  produced  by  molecular  cloning.  This  disease 
is  second  only  to  tuberculosis  as  a cause  of 
death  among  reportable  infectious  diseases.  It  is 
extremely  difficult  to  cultivate  the  causative 
agents.  Hepatitis  A has  a good  chance  of  being 
the  first  human  viral  disease  for  which  the  in- 
itial preparation  of  experimental  vaccine  will  in- 
volve molecular  cloning.  A vaccine  against  hepa- 
titis B,  made  from  the  blood  of  chronic  carriers, 


'For  other  aspects  of  vaccine  production  see:  Office  of  Technol- 
ogy .Assessment,  U.S.  Congress,  Working  Papers,  The  Impacts  of 
Genetics,  \ol.  2.  (Springfield,  Va.:  X'ational  Technical  Information 
Service,  1981). 

-Ibid. 


Figure  23. — Recombinant  DNA  Strategy  for  Making 
Foot-and-Mouth  Disease  Vaccine 


Growing  E.  coli  bacteria  may  produce  VPs  for  use  as  vaccine 
for  foot-and-mouth  disease.  No  virus  or  infectious  RNA  is 
produced  by  the  harmless  bacteria  strain. 


*VPs  is  the  protein  from  the  shell  of  the  virus,  which  can  act 
as  a vaccine  for  immunizing  livestock  against  foot-and- 
mouth  disease.  The  idea  outlined  above  is  to  make  this  VPs 
protein  without  making  any  virus  or  infectious  RNA. 

SOURCE:  Office  of  Technology  Assessnnent. 

is  in  the  testing  stage,  but  cloning  is  being  in- 
vestigated as  a better  source  of  an  appropriate 
antigen.  The  causative  agent  for  a third  form  of 
hepatitis  has  not  even  been  identified.  Since  at 
least  16  million  U.S.  citizens  are  estimated  to  be 
at  high  risk  of  contracting  hepatitis,  there  is 
keen  interest  in  the  development  of  vaccines 
among  academic  and  industrial  researchers.® 


’Ibid. 


70  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


More  hypothetically,  molecular  cloning  may 
lead  to  three  other  uses  of  antigens  as  well:  vac- 
cination against  parasites,  such  as  malaria  and 
hookworm  (see  Tech.  Note  7,  p.  80.);  immuniza- 
tion in  connection  with  cancer  treatment;  and 
counteracting  abnormal  antibodies,  which  are 
made  against  normal  tissues  in  the  so-called 
"autoimmune  diseases,”  such  as  multiple  sclero- 
sis. (See  Tech.  Note  8,  p.  81.) 

INTERFERONS 

Interferons  are  glycoproteins  normally  made 
by  a variety  of  cells  in  response  to  viral  infec- 
tion. All  interferons  (see  Tech.  Note  9,  p.  81)  can 
induce  an  antiviral  state  in  susceptible  cells.  In 
addition,  interferon  has  been  found  to  have  at 
least  15  other  biochemical  effects,  most  of 
which  involve  other  elements  of  the  immune 
system. 

Promising  preliminary  studies  have  sup- 
ported the  use  of  interferon  in  the  treatment  of 
such  viral  diseases  as  rabies,  hepatitis,  varicella- 
zoster  (shingles),  and  various  herpes  infections. 
To  date,  the  effect  of  interferon  has  been  far 
more  impressive  as  a prophylactic  than  as  a 
therapeutic  agent.  The  interferon  produced  by 


Genentech,  for  example,  has  been  shown  to  pro- 
tect squirrel  monkeys  from  infection  by  the  le- 
thal myocarditis  virus.  Once  interferon  is  avail- 
able in  quantity,  large-scale  tests  on  human  pop- 
ulations can  be  conducted  to  confirm  its  ef- 
ficacy in  man. 

Several  production  techniques  are  being  ex- 
plored. (See  Tech.  Note  10,  p.  81.)  Extraction  of 
interferon  from  leukocytes  (white  blood  cells), 
the  current  method  of  choice,  may  have  to  com- 
pete with  tissue  culture  production  as  well  as 
rDNA.  (See  table  5.) 

Recombinant  DNA  is  widely  regarded  as  the 
key  to  mass  production  of  interferons,  and 
important  initial  successes  ha\e  already  been 
achieved.  Each  of  the  four  major  biotechnology 
companies  is  working  on  improved  production 
methods,  and  all  have  reported  some  success. 

An  enormous  amount  remains  to  he  learned 
about  the  interferon  system.  It  now  appears 
that  the  interferons  are  simply  one  of  many 
families  of  molecules  involved  in  |)hvsiological 
regulation  of  response  to  disease.  Only  now 
have  molecular  biology  and  genetics  made  their 
study— and  perhaps  their  use— possible. 


Table  5.— Summary  of  Potential  Methods  for  Interferon  Production 


Means  of  production 

Types  of 

interferon 

produced 

Potential 

for 

scale-up 

Present  projected 
($/10®  units) 

Problems 

Potential  for 
improvement 

“Buffy  coat”  leukocytes 

leukocyte,  95% 
fibroblast,  5% 

No 

50  — 

— lack  of  scale-up 
—pathogen  contamination 

—minimal 

Lymphoblastoid  cells 

leukocyte,  80% 
fibroblast,  20% 

Yes 

— =25 

— poor  yields 

—cells  derived  from  tumor 

— improved  yields 
—expression  of 
fibroblast 
interferon 

Fibroblasts 

fibroblast 

Yes 

43-200  =1-10 

—cell  culture 
—economic  competition 
with  recombinant  DNA 

— improved  yields 

— improved  cell- 
culture 
technology 

—expression  of 
leukocyte-type 
interferon 

Recombinant  DNA 

leukocyte  or 
fibroblast 

Yes 

— =1-10 

—does  not  produce 
interferon 

— improved  yields 

— in  vitro  drug  stability 
— pooryieids  —modified 

interferons 

—drug  approval 
—possible  economic 
competition  with  fibroblast 
cell  production 


SOURCE:  Office  of  Technology  Assessment. 


Ch.4 — The  Pharmaceutical  Industry  • 71 


rhe  interferons  are  presently  recei\  ing  atten- 
tion largely  because  studies  in  Sweden  and  the 
L'nited  States  stimulated  the  appropriation  of 
$5.4  million  hy  the  American  Cancer  Society 
(.AC'S)  for  e.xpanded  clinical  trials  in  the  treat- 
ment of  cancer.  That  commitment  hy  the  non- 
profit AC'S— the  greatest  hy  far  in  its  history— 
was  followed  hy  a boost  in  NIH  funding  for  in- 
terferon research  from  $7.7  million  to  $19.9 
million  for  fiscal  year  1980.  Much  of  the  cost  of 
interferon  reseai'ch  is  allotted  to  procuring  the 
glycopeptide.  Initially,  the  ACS  bought  40  billion 
units  of  leukocyte  interferon  from  the  Finnish 
Red  Ca'oss  for  $50  per  million  units.  In  March 
1980,  Warner-Lambert  was  awarded  a contract 
to  supph’  the  National  C'ancer  Institute  (N'Cl) 
with  50  billion  units  of  leukocyte  interferon 
within  the  ne.xt  2 years  at  an  a\erage  price  of 
$18  per  million  units.  \'C4  is  also  planning  to 
purchase  50  billion  units  each  of  fibroblast  and 
lymphohlastoid  interferons. 

The  bulk  of  the  \IH  funding  is  included  in 
NCI’s  new  Biological  Response  Modifier  (BRM) 
program— interferon  accounts  for  $13.9  million 
of  the  $34.1  million  allocated  for  BRM  work  in 
fiscal  year  1980.  (NCI  expenditures  on  inter- 
feron in  1979  were  $2.6  million,  19  percent  of 
the  amount  budgeted  for  1980.)  Other  impor- 
tant elements  of  that  BRM  program  concern 
immunoproteins  known  as  lymphokines  and 
thymic  hormones,  for  which  molecular  genetics 
has  major  implications.  The  program  is  aimed  at 
identifying  and  testing  molecules  that  control 
the  acth’ities  of  different  cell  types. 

LYMPHOKINES  AND  CYTOKINES 

Lymphokines  and  cytokines  are  regulatory 
molecules  that  have  begun  to  emerge  from  the 
obscure  fringes  of  immunology  in  the  past  10 
years.  (Interferon  is  generally  considered  a lym- 
phokine  that  has  been  characterized  sufficiently 
to  deserve  independent  status.) 

Lymphokines  are  biologically  active  soluble 
factors  produced  by  w hite  blood  cells.  Studied 
in  depth  only  within  the  last  15  years,  they  are 
being  implicated  at  virtually  ev'ery  stage  in  the 
complex  series  of  events  that  make  up  the  im- 
mune response.  They  now'  include  about  100 
different  compounds.  Cytokines,  w'hich  have  ef- 


fects similar  to  lymphokines,  include  several 
compounds  associated  with  the  thymus  gland, 
referred  to  as  thymic  hormones.'* 

In  1979,  the  BRM  subcommittee  concluded 
that  se\  eral  of  these  agents  probably  have  great 
potential  for  cancer  treatment.  Nevertheless, 
adeciuate  quantities  for  laboratory  and  clinical 
testing  of  many  of  them  will  probably  not  he 
a\  ailahle  until  the  problems  of  producing  glyco- 
proteins by  molecular  cloning  are  overcome.  No 
system  is  currently  a\ailahle  for  the  industrial 
production  of  glycoproteins,  although  yeasts 
may  [)ro\e  to  he  the  most  useful  micro-orga- 
nisms. 

ANTIBODIES 

Antibodies  are  the  best  known  and  most  ex- 
ploited protein  components  of  the  immune  sys- 
tem. I’ntil  recently,  all  antibodies  were  obtained 
from  the  blood  of  humans  or  animals;  and  they 
were  often  impure.  Within  the  past  5 years, 
however,  it  has  become  possible  to  produce  an- 
tibodies from  cells  in  culture,  and  to  achieve 
levels  of  purity  previously  unattainable.  As  with 
prex’ious  adxances  in  antibody  technology,  re- 
searchers are  examining  ways  to  put  this  new 
le\el  of  purity  to  use.  There  have  been  hun- 
dreds, if  not  thousands,  of  examples  of  new 
diagnostic  and  research  methods,  new  methods 
of  purification,  and  new  therapies  published 
within  the  first  3 years  that  the  technique  has 
been  available.  (See  Tech.  Note  11,  p.  81.) 

This  high  level  of  purity  was  attained  by  the 
development  of  monoclonal  antibodies.  These 
antibodies  that  recognize  only  one  kind  of  anti- 
gen were  the  unanticipated  fruit  of  fundamen- 
tal immunological  research  conducted  by  Drs. 
Caesar  Milstein  and  Georges  Kohler  at  the  Med- 
ical Research  Council  in  England  in  1975.  They 
fused  two  types  of  cells— myeloma  and  plasma- 
spleen  cells— to  form  hybridomas  that  produce 
the  monoclonal  antibodies.  (See  Tech.  Note  12, 
p.  81.)  Not  only  are  the  antibodies  specific,  but 
because  the  hybridomas  can  be  grown  in  mass 
culture,  a virtually  limitless  supply  is  available. 

The  most  immediate  medical  application  for 
monoclonal  antibodies  lies  in  diagnostic  testing. 

■'For  40  of  the  best  characterized  cytokines,  see  footnote  1,  p. 
69. 


72  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Over  the  past  20  years,  large  segments  of  the 
diagnostic  and  clinical  laboratory  industries 
have  sprung  up  to  detect  and  quantify  particu- 
lar substances  in  specimens.  Because  monoclon- 
al antibodies  are  so  specific,  hybridomas  seem 
certain  to  replace  animals  as  the  source  of  anti- 
bodies for  virtually  all  diagnosis  and  monitor- 
ing. Tbeir  use  will  not  only  improve  tbe  accu- 
racy of  tests  and  decrease  development  costs, 
but  should  result  in  a more  uniform  product. 

Today,  such  assays  are  used  to: 

• determine  hormone  levels  in  order  to 
assess  the  proper  functioning  of  an  endo- 
crine gland  or  the  inappropriate  produc- 
tion of  a hormone  by  a tumor; 

• detect  certain  proteins,  tbe  presence  of 
which  has  been  found  to  correlate  with  a 
tumor  or  with  a specific  prenatal  condition; 

• detect  the  presence  of  illicit  drugs  in  a per- 
son’s blood,  or  monitor  the  blood  or  tissue 
level  of  a drug  to  ensure  tbat  the  dosage 
achieves  a therapeutic  level  without  ex- 
ceeding the  limits  that  could  cause  toxic  ef- 
fects; and 

• identify  microbial  pathogens. 

The  extent  of  the  use  of  antibodies  and  the 
biochemical  properties  that  they  can  identify  is 
suggested  by  table  6.  No  one  assay  constitutes  a 
major  market,  and  short  product  lifetime  has 
been  characteristic  of  this  business. 

Other  applications  of  monoclonal  antibodies 
include: 

• the  improvement  of  the  acceptance  of  kid- 
ney (and  other  organ)  transplants  by  injec- 
tion of  tbe  recipient  with  antibodies  against 
certain  antigens; 

• passive  immunization  against  an  antigen  in- 
volved in  reproduction,  as  a reversible  im- 
munological approach  to  contraception. 

• localizing  tumors  with  tumor-specific  anti- 
bodies (see  Tecb.  Note  13,  p.  81);  and 

• targeting  cancer  cells  with  antibodies  tbat 
bave  anticancer  chemicals  attached  to 
them. 


Enzymes  and  other  proteins 

ENZYMES 

Enzymes  are  involved  in  virtually  every  bio- 
logical process  and  are  well-understood.  Ne\'er- 
theless,  despite  tbeir  potency,  versatility,  and 
diversity,  they  play  a small  role  in  the  practice 
of  medicine  today.  Therapeutic  enzymes  ac- 
counted for  American  sales  of  about  $70  million 
(wholesale)  in  1978,  but  one-balf  of  those  sales 
involved  the  blood-plasma-derived  coagulation 
factors  used  to  treat  hemophilia.  Although  the 
figure  is  difficult  to  estimate,  the  total  numher 
of  patients  receiving  any  type  of  enzyme  ther- 
apy in  1980  probably  does  not  exceetl  v50,000. 

Enzymes  cannot  be  synthesized  by  con\en- 
tional  chemistry.  Almost  all  those  present  1\' 
employed  in  medicine  are  extracted  from 
human  blood,  urine,  or  organs,  or  are  produced 
by  micro-organisms.  Already  the  possibility  of 
using  rDNA  clones  as  the  source  of  enzymes— 
primarily  to  reduce  the  cost  of  i)roduction— is 
being  explored. 

However,  problems  associated  with  the  use  of 
nonhuman  enzymes  (such  as  immune  and  feb- 
rile responses)  and  the  scarcity  of  human  en- 
zymes, have  hindered  research,  de\('lopment, 
and  clinical  exploitation  of  enzyiiuvs  foi-  thei’- 
apeutic  purposes.  Today,  the  ex|)(>rimental  ge- 
netic technologies  of  rDNA  and  somatic  ('('ll  fu- 
sion and  culture  open  the  only  ('oncei\ able 
routes  to  relatively  inexpensi\('  [H'odiu'tion  of 
compatible  human  enzynies. 

The  genetic  engineering  of  enzymes  is  |)roh- 
ably  tbe  best  example  of  a dilemma  that  ham- 
pers the  exploitation  of  rDNA:  Without  a clinical 
need  large  enough  to  justify  the  iincstmenl, 
there  is  no  incenti\  e to  produce  a |)roduct:  yet 
without  adequate  supplies,  th(!  th('rapeutic  pos- 
sibilities cannot  be  in\  estigated,  I he  substances 
that  break  this  cycle  will  probably  he  those  that 
are  already  produced  in  (|uantity  from  tiatural 
tissue. 

The  only  enzymes  administered  today  .ire 
given  to  hemophiliacs— and  tlu'v  ai-e  .iclu.ill\ 


Ch.  4— The  Pharmaceutical  Industry  • 73 


Table  6.— Immunoassays 


Analgesics  and  narcotics 
Anileridine 
Antipyrine 
Codeine 
Etorphine 
Fentanyl 
Meperidine 
Methadone 
Morphine 
Pentazocine 
Antibiotics 
Amikacin 
Chloramphenicol 
Clindamycin 
Gentamicin 
Isoniazid 
Penicillin 
Sisomycin 
Tobramycin 
Anticonvulsants 
Clonazepam 
Phenytoin 
Primidone 

Anti-inflammatory  agents 
Colchicine 
Indomethacin 
Phenyibutazone 
Antineoplastic  agents 
Adriamycin 
Bleomycin 
Daunomycin 
Methotrexate 
Bronchodilators 
Theophylline 
Cardiovascular  drugs 
Cardiac  glycosides 
Acetylstrophanthidin 
Cedilanid 
Deslanoside 
Digitoxin 
Digoxin 
Gitoxin 


Hallucinogenic  drugs 
Mescaline 

Tetrahydrocannabinol 
Hypoglycemic  agents 
Butylbiguanide 
Glibenclamid 
Insecticides 
Aldrin 
DDT 
Dieldrin 
Malathion 

Narcotic  antagonists 
Cyclazocine 
Naloxone 
Peptide  hormones 
Angiotensin 
Anterior  pituitary 
Bradykinin 
Gastric 
Hypothalamic 
Intestinal 
Pancreatic 
Parathyroid 
Posterior  pituitary 
Thyroid  (calcitonin) 

Plant  hormones 
lndole-3-acetic  acid 
Gibberelilic  acid 
Polyamines 
Spermine 
Prostaglandins 
Sedatives  and 
tranquilizers 
Barbituarates 
Barbital 
Pentobarbital 
Phenobarbital 
Chlordiazepoxide 
Chlorpromazine 
Desmethylimipramine 
Diazepam  and 
N-desmethyIdiazepam 


Methyl  digoxin 
Ouabain 
Proscillaridin 
Dihydroergotamine 
Propranolol 
Quinidine 
CNS  stimulants 
Amphetamine 
Benzoyl  ecgonine 
(cocaine  metabolite) 
Methamphetamine 
Pimozide 
Diuretics 
Bumetanide 
Hallucinogenic  drugs 
Bile  acid  conjugates 
Cholylglycine 
Cholyltaurine 
Catecholamines 
Epinephrine 
Norepinephrine 
Tyramine 
Fibrinopeptides 
Fibrinopeptide  A 
Fibrinopeptide  B 
Indolealkylamines 
Melatonin 
Serotonin 
Insect  hormones 
Ecdysone 
Nucleosides  and 
nucleotides 
Cyclic  AMP 
Cyclic  GMP 
N*-Dimethylguanosine 
7-Methylguanosine 
Pseudouridine 
Thymidine 
Glutethimide 
Methaqualone 


Steroid  hormones 
Skeletal  muscle  relaxants 
d-Tubocurarine 
Synthetic  peptides 
DDAVP 
Saralasin 
Synthetic  steroids 
Anabolic  steroids 
Trienbolone  acetate 
Androgens 

Fluoxymesterone 

Estrogens 

Diethylstilbestrol 

Ethinylestradiol 

Mestranol 

Glucocorticoids 

Dexamethasone 

Methylprednisolone 

Prednisolone 

Prednisone 

Metyrapone 

Progestins 

Medroxyprogesterone 
acetate 
Norethindrone 
Norethisterone 
Norgestrel 
Toxins 
Aflatoxin  B, 

Genistein 

Nicotine  and  metabolites 
Ochratoxin  A 
Paralytic  shellfish  poison 
Thyroid  hormones 
Thyroxine 
Triodothyronine 
Vitamins 
Vitamin  B12 
Vitamin  D 


SOURCE:  "Immunoassays  of  Drugs— Comprehensive  Immunology."  Immunal  Pharmacology.  Hadden  Caffey  (ed.)(New  York:  Plenum  Press,  1977),  p.  325. 


proenzymes,  which  are  converted  to  active  en- 
zymes in  the  body  when  needed.  The  most  com- 
mon agents  are  called  Factor  \TII  and  Factor  IX, 
which  are  found  in  serum  albumin  and  are  cur- 
rently extracted  from  human  blood  plasma. 
Hemophilia  .A  and  Hemophilia  B— accounting  for 
over  90  percent  of  all  major  bleeding  disor- 
ders—are  characterized  by  a deficiency  of  these 
factors.  Supplies  of  the  proenz\anes  will  exceed 
demand  w ell  beyond  1980  if  the  harvesting  and 
processing  of  plasma  continues  as  it  has.  Never- 
theless, the  risk  of  hepatitis  associated  with  the 


use  of  human  plasma-derived  products  is  ex- 
tremely high.  One  recent  study  found  chronic 
hepatitis  in  a significant  percentage  of  asymp- 
tomatic patients  treated  with  Factor  VIII  and 
Factor  IX. 

The  plasma  fractionation  industry,  which 
produces  the  proenzymes,  is  currently  faced 
with  excess  capacity,  intense  competition,  high 
plasma  costs,  and  tight  profit  margins.®  The  cost 
and  availability  of  any  one  plasma  protein  is 

=For  details  of  the  factors  governing  the  industry,  see  footnote  1, 
p.  69. 


74  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


coupled  to  the  production  of  the  others.  Hence, 
the  industry  would  still  have  to  orchestrate  the 
production  of  the  other  proteins  even  if  just  one 
of  them,  such  as  Factor  VIII,  becomes  a target 
for  biological  production. 

Another  enzyme,  urokinase,  has  been  tar- 
geted for  use  in  removing  unwanted  blood  clots, 
which  lead  to  strokes,  myocardial  infarctions, 
and  pulmonary  emboli.  Currently,  the  drug  is 
either  isolated  from  urine  or  produced  in  tissue 
culture.  (See  Tech.  Note  14,  p.  81.) 

Urokinase  is  thus  far  the  only  commercial 
therapeutic  product  derived  from  mammalian 
cell  culture.  Nevertheless,  some  calculations 
suggest  that  production  by  E.  coU  fermentation 
would  have  economic  advantages.  The  costs  im- 
plicit in  having  to  grow  cells  for  30  days  on  fetal 
calf  serum  (or  its  equivalent)  or  in  having  to  col- 
lect and  fractionate  urine— as  reflected  in  uroki- 
nase’s market  price  ($150/mg  at  the  manufac- 
turer’s level)— should  be  enough  incentive  to  en- 
courage research  into  its  production.  In  fact,  in 
April  1980,  Abbott  Laboratories  disclosed  that 
E.  coli  had  been  induced  to  produce  urokinase 
through  plasmid-borne  DNA. 

The  availability  of  urokinase  might  be  guar- 
anteed by  the  new  genetic  technologies,  but  its 
use  is  not.  For  a variety  of  reasons,  the  Amer- 
ican medical  community  has  not  accepted  the 
drug  as  readily  as  have  the  European  and  Japa- 
nese communities.  Studies  to  establish  the  use 
of  urokinase  for  deep  vein  thrombosis,  for  ex- 
ample, are  now  being  conducted  almost  exclu- 
sively in  Europe.® 

OTHER  PROTEINS 

In  addition  to  the  proteins  and  polypeptides 
already  mentioned,  the  structural  proteins, 
such  as  the  collagens  (the  most  abundant  pro- 
teins in  the  body),  elastins  and  keratins  (the 
compounds  of  extracellular  structures  like  hair 
and  connective  tissue),  albumins,  globulins,  and 
a wide  variety  of  others,  may  also  be  susceptible 
to  genetic  engineering.  Structural  proteins  are 
less  likely  to  be  suitable  for  molecular  genetic 
manipulations:  On  the  one  hand,  their  size  and 


®For  additional  information  about  how  urokinase  came  to  play  a 
role  in  therapy,  see  footnote  1,  p.  69. 


complexity  exceed  the  synthetic  and  analytic 
capabilities  that  will  be  available  in  the  next  few 
years;  on  the  other,  either  their  use  in  medicine 
has  yet  to  be  established  or  material  derived 
from  animals  appears  adequate,  as  is  the  case 
with  collagen,  for  which  uses  are  emerging. 

Plasma,  the  fluid  portion  of  the  blood,  con- 
tains about  10  percent  solids,  most  of  which  are 
proteins.  During  World  War  II,  a simple  pro- 
cedure was  developed  to  separate  the  various 
components.  It  is  still  used  today. 

Serum  albumin  is  the  smallest  of  the  main 
plasma  proteins  but  it  constitutes  about  half  of 
plasma’s  total  mass.  Its  major  therapeutic  use  is 
to  reverse  the  effects  of  shock. ^ It  is  a reason- 
able candidate  for  molecular  cloning,  although 
its  relatively  high  molecular  weight  complicates 
purification,  and  its  commercial  \alue  is  rela- 
tively low.  The  market  value  of  normal  serum 
albumin  is  approximately  $3/g,  hut  the  \’olume 
is  such  that  domestic  sales  exceed  $150  million. 
Including  exports,  annual  production  is  in  th(* 
range  of  100,000  kg. 

Normal  serum  albumin  for  treating  shock  is 
already  regarded  as  too  expensive  compared 
with  alternative  treatments,  to  expand  its  use 
would  require  a lower  price.  On  the  other  hand, 
the  Federal  Government— and  especially  the  De- 
partment of  Defense— might  disregai'd  the  im- 
mediate economic  prospects  and  conclude'  that 
having  a source  of  human  serum  albumin  tliat 
does  not  depend  on  payments  to  blood  donors 
might  be  in  the  national  intei'est.  Since*  many  na- 
tions import  serum  albumin,  proeku'ts  ele*ri\e*el 
from  molecular  cloning  e;e)ulel  he  expoi  te*el. 

Serum  albumin  is  presently  the  prineipal 
product  of  blood  plasma  fractie)nation,  a e-hange 
in  the  way  it  is  manufactureel  we)ulel  signifie'ant- 
ly  affect  that  industry,  lieeiau.sei  a numhe*r  eif 
other  products  (such  as  cle)tting  fae'tors)  are  al.se) 
derived  from  fractie)natie)ii,  a growth  in  the* 
need  for  plasma-elei’iveel  albumin  e’e)ulel  ha\ c a 
significant  impact  e)ii  the  a\ailahility  anel  the* 
cost  of  these  hypre)elucts. 


Tor  a cietailc-d  cli.snission  ol  (he  cosIn  and  bcin-lils  ol  .ilbtj 

min  and  the  striiclur(M)l  iIh>  indu.sirv.  M'c  loolnoic  I p (i!i 


Ch.4 — The  Pharmaceutical  Industry  • 75 


Antibiotics 

Antimicrobial  agents  for  the  treatment  of  in- 
fectious diseases  ha\e  been  the  largest  selling 
prescription  pharmaceuticals  in  the  world  for 
the  past  three  decades.  Most  of  these  agents  are 
antibiotics— antimicrobials  naturally  produced 
by  micro-organisms  rather  than  by  chemical 
synthesis  or  by  isolation  from  higher  organisms. 
However,  one  major  antibiotic,  chlorampheni- 
col—originally  produced  by  a micro-organism, 
is  now  synthesized  by  chemical  methods.  The 
field  of  antibiotics,  in  fact,  pro\  ides  most  of  the 
precedent  for  employing  microbial  fermenta- 
tion to  produce  useful  medical  substances.  The 
L'nited  States  has  been  prominent  in  their 
development,  production,  and  marketing,  with 
the  result  that  .American  companies  account  for 
about  half  of  the  roughly  S5  billion  worth  of  an- 
timicrobial agents  sold  worldwide  each  year. 
7'he  .American  market  share  has  been  growing 
as  new  antibiotics  are  de\eloped  and  intro- 
duced e\erv  year. 

For  30  years,  high-yielding,  antibiotic-pro- 
ducing micro-organisms  ha\  e been  identified  by 
selection  from  among  mutant  strains.  Initially, 
organisms  producing  new  antibiotics  are  iso- 
lated by  soil  sampling  and  other  broad  screen- 
ing efforts.  They  are  then  cultured  in  the  lab- 
oratory, and  efforts  are  made  to  improx  e their 
productivity. 

Antibiotics  are  complex,  usually  nonprotein, 
substances,  which  are  generally  the  end  prod- 
ucts of  a series  of  biological  steps.  U'hile  knowl- 
edge of  molecular  details  in  metabolism  has 
made  some  difference,  not  a single  antibiotic 
has  had  its  complete  biosynthetic  pathway  eluci- 
dated. This  is  partly  because  there  is  no  single 
gene  that  can  be  isolated  to  produce  an  antibi- 
otic. However,  mutations  can  be  induced  within 
the  original  micro-organism  so  that  the  level  of 
production  can  be  increased. 

Other  methods  can  also  increase  production, 
and  possibly  create  new  antibiotics.  Microbial 
mating,  for  example,  which  leads  to  natural 
recombination,  has  been  widely  investigated  as 
a way  of  developing  vigorous,  high-yielding  an- 
tibiotic producers.  However,  its  use  has  been 
limited  by  the  mating  incompatibility  of  many 


industi'ially  important  higher  fungi,  the  pres- 
ence of  chromosomal  aberrations  in  micro-orga- 
nisms improved  by  mutation,  and  a number  of 
other  problems.  Furthermore,  natural  recom- 
bination is  most  ad\  antageous  when  strains  of 
extremely  diverse  origins  are  mated;  the  pro- 
prietary secrets  protecting  commercial  strains 
usually  j)revent  the  sort  of  divergent  "competi- 
tor” strains  most  likely  to  produce  vigorous 
hybrids  from  being  brought  together. 

The  technique  of  pi'otoplast  or  cell  fusion 
provides  a convenient  method  for  establishing  a 
recombinant  system  in  strains,  species,  and 
genera  that  lack  an  efficient  natural  means  for 
mating.  For  example,  as  many  as  four  strains  of 
the  antibiotic-producing  bacterium  Streptomy- 
ces  have  been  fused  together  in  a single  step  to 
yield  recombinants  that  inherit  genes  from  four 
parents.  The  technique  is  applicable  to  nearly 
all  antibiotic  producers.  It  will  help  combine  the 
benefits  developed  in  divergent  lines  by  muta- 
tion and  selection. 

In  addition,  researchers  have  compared  the 
quality  of  an  antibiotic-producing  fungus,  Ceph- 
alosporium  acremonium,  produced  by  mating  to 
one  produced  by  protoplast  fusion.  (See  Tech. 
Note  15,  p.  82.)  They  concluded  that  protoplast 
fusion  was  far  superior  for  that  purpose.  What 
is  more,  protoplast  fusion  can  give  rise  to  hun- 
dreds of  recombinants— including  one  isolate 
that  consistently  produced  the  antibiotic  ceph- 
alosporin C in  40  percent  greater  yield  than  the 
best  producer  among  its  parents— without  los- 
ing that  parent  strain’s  rare  capacity  to  use  in- 
organic sulfate,  rather  than  expensive  methio- 
nine, as  a source  of  sulfur.  It  also  acquired  the 
rapid  growth  and  sporulation  characteristics  of 
its  less-productive  parent.  Thus,  desirable  at- 
tributes from  different  parents  were  combined 
in  an  important  industrial  organism  that  had 
proved  resistant  to  conventional  crossing. 

Even  more  significant  are  the  possibilities  for 
preparation  by  protoplast  fusion  between  dif- 
ferent species  or  genera  of  hybrid  strains, 
which  could  have  unique  biosynthetic  capaci- 
ties. One  group  is  reported  to  have  isolated  a 
novel  antibiotic,  clearly  not  produced  by  either 
parent,  in  an  organism  created  through  fusion 
of  actinomycete  protoplasts.  (See  Tech.  Note  16, 


76  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


p.  82.)  The  value  of  protoplast  fusion,  therefore, 
lies  in  potentially  broadening  the  gene  pool. 

Protoplast  fusion  is  genetic  recombination  on 
a large  scale.  Instead  of  one  or  a few  genes  be- 
ing transferred  across  genus  and  species  bar- 
riers, entire  sets  of  genes  can  be  moved.  Success 
is  not  assured,  however;  a weakness  today  is  the 
inherited  instability  of  the  “fused”  clones.  The 
preservation  of  traits  and  long-range  stability 
has  yet  to  be  resolved.  Furthermore,  it  seems 
that  one  of  the  most  daunting  problems  is 
screening— determining  what  to  look  for  and 
how  to  recognize  it.  (See  Tech.  Note  17,  p.  82.) 

Recombinant  DNA  techniques  are  also  being 
examined  for  their  ability  to  improve  strains. 
Many  potentially  useful  antibiotics  do  not  reach 
their  commercial  potential  because  the  micro- 
organisms cannot  be  induced  to  produce  suffi- 
cient quantities  by  traditional  methods.  The  syn- 
thesis of  certain  antibiotics  is  controlled  by 
plasmids,  and  it  is  believed  that  some  plasmids 
may  nonspecifically  enhance  antibiotic  produc- 
tion and  excretion. 

It  may  also  be  possible  to  transfer  as  a group, 
all  the  genes  needed  to  produce  an  antibiotic 
into  a new  host.  However,  increasing  the  num- 
ber of  copies  of  critical  genes  by  phage  or  plas- 
mid transfer  has  yet  to  be  achieved  in  antibiotic- 
producing  organisms  because  little  is  known  of 
the  potential  vectors.  The  genetic  systems  of 
commercial  strains  will  have  to  be  understood 
before  the  newer  genetic  engineering  ap- 
proaches can  be  used.  Genetic  maps  have  been 
published  for  only  3 of  the  24  or  more  indus- 
trially useful  bacteria. 

Since  2,000  of  the  2,400  known  antibiotics  are 
produced  by  Streptomyces,  that  is  the  genus  of 
greatest  interest  to  the  pharmaceutical  indus- 
try. Probably  every  company  conducting  re- 
search on  Streptomyces  is  developing  vectors, 
but  little  of  the  industrial  work  has  been  re- 
vealed to  date. 


Nonprotein  pharmaceuticals 

In  both  sales  and  quantity,  over  80  percent  of 
the  pharmaceuticals  produced  today  are  not 
made  of  protein.  Instead,  they  consist  of  a varie- 


ty of  organic  chemical  entities.  These  drugs,  ex- 
cept for  antibiotics,  are  either  extracted  from 
some  natural  plant  or  animal  source  or  are  syn- 
thesized chemically. 

Some  of  the  raw  materials  for  pharmaceuti- 
cals are  also  obtained  from  plants;  micro-orga- 
nisms are  then  used  to  convert  the  material  to 
useful  drugs  in  one  or  two  enzymatic  steps. 
Such  conversions  are  common  for  steroid  hor- 
mones. 

In  1949,  when  cortisone  was  found  to  he  a 
useful  agent  in  the  treatment  of  arthritis,  the 
demand  for  the  drug  could  not  be  met  since  no 
practical  method  for  large-scale  production  ex- 
isted. The  chemical  synthesis  was  complicated 
and  very  expensive.  In  the  early  and  micl-195()'s, 
many  investigators  reported  the  microbial 
transformation  of  several  intermediates  to  com- 
pounds that  corresponded  to  the  chemical  syn- 
thetic scheme.  By  saving  many  chemical  steps 
and  achieving  higher  yields,  manufactui'crs 
managed  to  reduce  the  price  of  stei'oids  to  a 
level  where  they  were  a marketable  commodi- 
ty. A conversion  of  progesterone,  for  e.\ampU>, 
dropped  the  price  of  cortisone  from  $200  to 
$6/g  in  1949.  Through  fui'thei’  impro\'(Mnents, 
the  price  dropped  to  less  than  $l/g.  The  1980 
price  is  $0.46/g. 

Developments  based  on  genetic;  te('hni(|ucs  to 
increase  the  production  and  secrcUioii  of  kc\  en- 
zymes could  substantially  improxc;  the  econom- 
ics of  some  presently  inefficient  pi’ocesses.  Cur- 
rently, assessments  are  being  ('ariied  out  by 
various  companies  to  determine'  which  of  the 
many  nonprotein  phai’macxnitieals  c'an  hc'  man- 
ufactured more  readily  oi'  more  ('conomieally 
by  biological  means. 

Approximately  90  perccMit  of  the*  pharmaceu- 
ticals used  in  the  treatiimnt  of  hypei  ten.sion  ai'c 
obtained  from  plants,  as  well  as  are  miscel- 
laneous cardio\asculai’  drugs.  Morphine  .md 
important  \asodilators  are  obtained  from  tlie 
opium  poj)py,  Papaver  sotttniferiim.  All  these 
chemical  substances  arc;  produec'd  by  a series  nl 
enzymes  that  ai’e  codcnl  h\'  con-esponding  ^;enes 
in  the  whole  plant.  The'  long-term  possihilitv 
(over  10  years)  of  using  fermentation  methods 
will  depend  on  idcMitifving  the  important  ^enes 


Ch.  4— The  Pharmaceutical  Industry  • 77 


The  genes  that  are  transferred  from  plant  to 
bacteria  must  ob\  iously  be  ileterminecl  on  a 
case-bv-case  basis.  The  case  study  on  acetamino- 
phen (the  acti\  e ingredient  in  analgesics  such  as 
Tylenol)  demonstrates  the  steps  in  such  a feasi- 
bility study.  (Seeapp.  l-.\.) 

'I'be  first  stej)  in  such  a study  is  to  detei'inine 
w hetber  and  w here  enzymes  e.xist  to  carry  out 
the  necessary  transformation  for  a given  prod- 
uct. .-\cetaminophen  for  instance,  can  be  made 
from  aniline,  a relati\ely  ine.\pensi\e  starting 
material.  The  two  necessary  enzymes  can  be 


found  in  several  fungi.  Either  the  enzymes  can 
be  isolated  and  used  directly  in  a two-step  con- 
version or  the  genes  for  both  enzymes  can  be 
transferred  into  an  organism  that  can  carry  out 
the  entire  conversion  by  itself. 

(li\en  the  cost  assumptions  outlined  in  the 
case  study  and  the  assumptions  on  the  efficien- 
cy of  comerting  aniline  to  acetaminophen,  the 
cost  of  producing  the  drug  by  fermentation 
could  be  20  percent  lower  than  production  by 
chemical  synthesis. 


Impacts  

Genetic  technologies  can  help  pro\  ide  a \ arie- 
ty  of  pharmaceutical  products,  many  of  which 
ha\  e been  identified  in  this  report.  But  the  tech- 
nologies cannot  guarantee  how  a product  will 
he  used  or  even  whether  it  u ill  he  used  at  all. 
The  pharmaceuticals  discussed  ha\e  illustrated 
the  kinds  of  major  economic,  technical,  social, 
and  legal  constraints  that  u ill  play  a role  in  the 
application  of  genetic  technologies. 

Clearly,  the  major  direct  impacts  of  genetic 
technologies  will  be  felt  primarily  through  the 
type  of  products  they  bring  to  market.  Never- 
theless, each  new  pharmaceutical  will  offer  its 
own  spectrum  and  magnitude  of  impacts.  Tech- 
nically, genetic  engineering  may  lead  to  the  pro- 
duction of  growth  hormone  and  interferon  with 
equal  likelihood;  but  if  the  patient  population  is 
a thousandfold  higher  for  interferon,  and  if  its 
therapeutic-effect  is  to  alle\  iate  pain  and  lower 
the  cancer  mortality  rate,  its  impact  will  be  sig- 
nificantly greater. 

Many  hormones  and  human  proteins  cannot 
be  extensively  studied  because  they  are  still 
either  unax  ailable  or  too  expensiv'e.  Until  the 
physiological  properties  of  a hormone  are 
understood,  its  therapeutic  \ alues  remain  un- 
known. Recombinant  DNA  techniques  are  being 
used  to  overcome  this  circular  problem.  In  one 
laboratory,  somatostatin  is  being  used  as  a re- 
search tool  to  study  the  regulation  of  the  hor- 
monal milieu  of  burn  patients.  A single  experi- 
ment may  use  as  much  as  25  mg  of  the  hor- 


mone, which,  as  a product  of  solid  state  chem- 
ical synthesis,  costs  as  much  as  $12,000.  Re- 
ducing its  cost  would  allow  for  more  extensive 
research  on  its  physiological  and  therapeutic 
qualities. 

By  making  a pharmaceutical  available,  genet- 
ic engineering  can  have  two  types  of  impacts. 
First,  pharmaceuticals  that  already  have  med- 
ical promise  will  be  available  for  testing.  For  ex- 
ample, interferon  can  be  tested  for  its  efficacy 
in  cancer  and  viral  therapy,  and  human  growth 
hormone  can  be  evaluated  for  its  ability  to  heal 
wounds.  For  these  medical  conditions,  the  in- 
direct, societal  impact  of  applied  genetics  could 
be  widespread. 

Second,  other  pharmacologically  active  sub- 
stances that  have  no  present  use  will  be  avail- 
able in  sufficient  quantities  and  at  a low  enough 
cost  to  enable  researchers  to  explore  their  possi- 
bilities, thus  creating  the  potential  for  totally 
new  therapies.  Genetic  technologies  can  make 
available  for  example,  cell  regulatory  proteins,  a 
class  of  molecules  that  control  gene  activity  and 
that  is  found  in  only  minute  quantities  in  the 
body.  The  cytokines  and  lymphokines  typify  the 
countless  rare  molecules  involved  in  regulation, 
communication,  and  defense  of  the  body  to 
maintain  health.  Now,  for  the  first  time,  genetic 
technologies  make  it  possible  to  recognize,  iso- 
late, characterize,  and  produce  these  proteins. 

The  potential  importance  of  this  class  of  phar- 
maceuticals—the  new  cell  regulatory  mole- 


78  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


cules— is  underscored  by  the  fact  that  half  of 
the  22  active  INDs  for  new  molecular  entities 
that  have  been  rated  by  FDA  as  promising  im- 
portant therapeutic  gains  are  in  the  Metabolic 
and  Endocrine  Division,  which  oversees  such 
drugs.  It  is  reasonable  to  anticipate  that  they 
will  be  employed  to  treat  cancer,  to  prevent  or 
combat  infections,  to  facilitate  transplantation 
of  organs  and  skin,  and  to  treat  allergies  and 
other  diseases  in  which  the  immune  system  has 
turned  against  the  organism  to  which  it  belongs. 
(See  table  7.) 

At  the  very  least,  even  if  immediate  medical 
uses  cannot  be  found  for  any  of  these  com- 
pounds, their  indirect  impact  on  medical  re- 
search is  assured.  For  the  first  time,  almost  any 
biological  phenomenon  of  medical  interest  can 
be  explored  at  the  cellular  level  by  the  appli- 


Table  7.— -Diseases  Amenable  to  Drugs  Produced  by 
Genetic  Engineering  in  the  Pharmaceutical  Industry 


Disease  or  condition 

Drug  potentially  produced  by 
genetically  engineered  organism 

Diabetes^ 

Insulin 

Atherosclerosis 

Platelet-derived  growth  factor 
(PDGF) 

Virus  diseases 
Influenza 
Hepatitis 
Polio 
Herpes 
Common  cold 

Interferon 

Cancer 

Interferon 
Hodgkin’s  disease 
Leukemia 
Breast  cancer 

Anovulation 

Human  chorionic  gonadatropin 

Dwarfism^ 

Human  growth  hormone 

Pain 

Enkephalins  and  endorphins 

Wounds  and  burns 
Inflammation, 

Human  growth  hormone 

rheumatic  diseases^ 
Bone  disorders,  e.g., 

Adrenocorticotrophic  hormone 
(ACTH) 

Paget’s  disease^ 

Calcitonin  and  parathyroid 
hormone 

Nerve  damage 

Nerve  growth  factor  (NGF) 

Anemia,  hemorrhage 

Erythropoietin 

Hemophilia® 

Factor  VIII  and  Factor  IX 

Blood  clots® 

Urokinase 

Shock® 

Serum  albumin 

Immune  disorders 

Cytokines 

^Indicates  diseases  currently  treated  by  the  drugs  listed. 
SOURCE:  Office  of  Technology  Assessment. 


cation  of  available  scientific  tools.  These  new 
molecules  are  valuable  tools  for  dissecting  the 
structure  and  function  of  the  cell.  The  knowl- 
edge gained  may  lead  to  the  development  of 
new  therapies  or  preventive  measures  for 
diseases. 

The  increased  availability  of  new  \accines 
might  also  have  serious  consequences.  But  the 
extent  to  which  molecular  cloning  will  prox  ide 
useful  vaccines  for  intractable  diseases  is  still 
unknown.  For  some  widespread  diseases,  such 
as  amebic  dysentery,  not  enough  is  known 
about  the  interaction  between  the  micro-orga- 
nism and  the  patient  to  help  researchers  design 
a rational  plan  of  attack.  For  others,  such  as 
trachoma,  malaria,  hepatitis,  and  influenza, 
there  is  only  preliminary  experimental  ex  idence 
that  a useful  vaccine  could  he  produced.  (See 
table  8.)  To  date,  the  xaccine  that  is  most  likely 
to  have  an  immediate  impact  combats  foot-and- 
mouth  disease  in  veterinary  medicine.  Fhere  is 
little  doubt  however,  that  should  any  one  of  the 
vaccines  for  human  diseases  become  ax  ailahle, 
the  societal,  economic,  and  political  conse- 
quences of  a decrease  in  morbidity  and  mortali- 
ty would  be  significant.  Many  of  thesf'  diseases 
are  particularly  prevalent  in  less-dexeloped 
countries.  The  effects  of  dexeloping  xaccimvs 

Table  8.— Major  Diseases  for  Which  Vaccines 
Need  To  Be  Developed 


Parasitic  diseases 
Hookworm 
Trachoma 
Malaria 

Schistosomiasis 
Sleeping  sickness 
Viruses 
Hepatitis 
Influenza 

Foot-and-mouth  disease  (for  cloven-hoofed  animals) 
Newcastle  disease  virus  (for  poultry) 

Herpes  simplex 

Mumps 

Measles 

Common  cold  rhinoviruses 
Varicella-zoster  (shingles) 

Bacteria 
Dysentery 
Typhoid  fever 
Cholera 

Traveller’s  diarrhea 

SOURCE:  Office  of  Technology  Assessment 


Ch.  4— The  Pharmaceutical  Industry  • 79 


for  them  v\  ill  he  felt  on  an  international  scale 
and  w ill  in\  oK  e luindi'eds  of  millions  of  people. 

rhe  new  technologies  may  also  lower  the 
risks  of  \accine  production.  For  e.xample,  the 
FMD\  vaccine  produced  hv  (ienentech  is  con- 
structed out  of  17  of  the  20  genes  in  the  entire 
v irus— enough  to  confer  resistance,  hut  too  few 
to  dev  elop  into  a v iable  organism. 

I'he  new  technologv'  may  also  supplv  piiarma- 
ceuticals  with  effects  heyond  therajn'.  .At  least 
tw  o promise  ini[)acts  vv  ith  hroad  consequences: 
MSH  AC  FH  4-10  can  he  e.\[)ecled  to  he  used  on  a 
wide  scale  if  it  is  shown  to  improve  memory; 
and  homhesin  and  cholecystokinin  might  e.\- 
pand  the  appetite  suppression  market.  But  nei- 
ther of  these  compounds  has  yet  been  found  to 
he  useful.  U bile  genetic  technologies  may  pro- 
vide  large  suj)plies  of  the  diugs,  they  do  not 
guarantee  their  v alue. 

•Antibody -based  diagnostic  tests,  developed 
through  genetic  engineering,  may  eventually  in- 
clude early  warning  signals  for  cancer;  they 
should  he  able  to  recognize  any  one  of  the 
scores  of  cancers  that  cause  about  a half-million 
deaths  per  year  in  the  United  States.  If  anti- 
bodies prov  e successful  as  diagnostic  screening 
agents  to  predict  disease,  large-scale  screening 
of  the  population  can  occur,  accelerating  the 
trend  toward  preventiv  e medicine  in  the  United 
States. 

In  addition  to  drugs  and  diagnostic  agents, 
proteins  could  be  produced  for  laboratory  use. 
E.xpensive,  complex  media  such  as  fetal  calf 
serum  are  presently  required  for  growing  most 
mammalian  tissue  cells.  Genetic  cloning  could 
make  it  possible  to  synthesize  vital  constituents 
cheaply,  and  could  markedly  reduce  the  costs  of 
cell  culture  for  both  research  and  production. 
Ironically,  genetic  cloning  could  make  economi- 
cally competitive  the  very  technology  that  of- 
fers an  alternative  production  method  for  many 
drugs:  tissue  culture. 

Xevertheless,  the  mere  availability  of  a phar- 
macologically active  substance  does  not  ensure 
its  adoption  in  medical  practice.  Even  if  it  is 
shown  to  have  therapeutic  usefulness,  it  may 
not  succeed  in  the  marketplace.  Consumer  re- 
sistance limits  the  use  of  some  drugs.  The  Amer- 


ican aversion  to  therapies  that  rec|uire  frequent 
injection,  for  instance,  is  illustrated  by  the  opin- 
ion of  some  that  a drug  like  AC^TH  offers  few,  if 
any,  adv  antages  over  steroids. 

The  use  of  Atn  il  is  somewhat  greater  abroad 
than  in  the  United  States.  This  is  due  in  part 
because  physicians  in  other  cultui'es  make  far 
less  use  of  systemic  steroids  than  their  Amer- 
ican counterparts,  and  in  part  because  frequent 
injections  are  more  acceptable  hence  more  com- 
mon. Sales  of  ACnil  in  Great  Britain— with 
its  much  smaller  population— ecpial  American 
sales. 

■At  present,  the  need  for  injection  is  a far 
more  likely  deterrent  to  the  wider  use  of  AC  FH 
than  the  cost  of  the  drug  itself.  Keports  that  it 
can  he  ap()lied  by  nasal  spray  suggest  that  its 
use  may  grow.  Implantable  controlled-release 
dosages  may  also  become  available  within  the 
next  5 years.  Fhis  dependence  on  appropriate 
drug  delivery  mechanisms  may  lead  to  another 
line  of  research— increased  attempts  to  develop 
technologies  for  drug-delivery. 

As  new  pharmaceuticals  become  available, 
disrufjtion  can  be  expected  to  occur  in  the  sup- 
ply of  some  old  ones.  Pharmaceuticals  whose 
production  is  tied  to  the  production  of  others 
might  become  increasingly  expensive  to  pro- 
duce. Clotting  factors,  for  example,  are  ex- 
tracted with  other  blood  components  from 
plasma.  Nevertheless,  producing  any  of  the  14 
currently  approved  blood  plasma  products  by 
rDNA  would  reduce  the  incidence  of  hepatitis 
caused  by  contamination  from  natural  blood 
sources. 

Whether  new  pharmaceuticals  are  produced 
or  new  production  methods  for  existing  phar- 
maceuticals are  dev'ised,  future  sources  for  the 
drugs  may  change.  Currently,  the  sources  are 
div'erse,  including  many  different  plants,  nu- 
merous animal  organs,  various  tissue  culture 
cells,  and  a wide  range  of  raw  materials  used 
for  chemical  synthesis.  A massive  shift  to  fer- 
mentation would  narrow  the  selection.  The  im- 
pacts on  present  sources  can  only  be  judged  on 
a case-by-case  basis.  The  new  sources— micro- 
organisms and  the  materials  that  feed  them— 
offer  the  guarantee  that  the  raw  materials  won’t 
dry  up.  If  one  disappears,  another  can  be  found. 


80  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


Clearly,  there  is  no  simple  formula  to  identify 
all  the  impacts  of  applied  genetics  on  the  phar- 
maceutical industry.  Even  projections  of  eco- 
nomic impacts  must  remain  crude  estimates. 
Nevertheless,  the  degree  to  which  genetic  engi- 
neering and  fermentation  technologies  might 
potentially  account  for  drug  production  in  spe- 
cific categories  is  projected  in  appendix  I-B. 


Given  the  assumptions  described,  the  inimedi- 
ate  direct  economic  impact  of  using  genetic  ma- 
nipulation in  the  industry,  measured  as  sales, 
can  be  estimated  in  the  billions  of  dollars,  with 
the  indirect  impacts  (sales  for  suppliers,  savings 
due  to  decreased  sick  days,  etc.)  reaching 
several  times  that  value. 


Technical  notes 


1.  Many  hormones  are  simply  chains  of  amino  acids  (poly- 
peptides); some  are  polypeptides  that  have  been  mod- 
ified by  the  attachment  of  carbohydrates  (glycopep- 
tides).  Hormones  usually  trigger  events  in  cells  remote 
from  the  cells  that  produced  them.  Some  act  over 
relatively  short  distances— between  segments  in  the 
brain,  or  in  glands  closely  linked  to  the  brain,  others 
act  on  distant  sites  in  tissues  throughout  the  body. 

2.  For  peptides  about  30  AA  in  length,  the  cost  may  ap- 
proach $1  per  mg  as  the  volume  approaches  the  kilo- 
gram level— a level  of  demand  rarely  existing  today  but 
likely  to  be  generated  by  work  in  progress.  Today,  the 
cost  of  the  32  AA  polypeptide,  calcitonin,  which  is  syn- 
thesized chemically  and  marketed  as  a pharmaceutical 
product  by  Armour,  is  probably  in  the  range  of  $20  per 
mg,  since  the  wholesale  price  in  vials  containing  ap- 
proximately 0.15  mg  is  about  $85/mg.  (That  price  is  an 
educated  guess,  since  such  costs  are  closely  guarded 
secrets  and  since  the  price  of  a pharmaceutical  in- 
cludes so  many  variables  that  the  cost  of  the  agent 
itself  is  a small  consideration.) 

3.  In  addition  to  those  helped  by  the  National  Pituitary 
Agency,  another  100  to  400  patients  are  treated  with 
hGH  from  commercial  sources.  The  commercial  price 
is  approximately  $15  per  unit  (roughly  $30/mg).  The 
production  cost  at  the  National  Pituitary  Agency  is 
about  $0. 75/unit  ($1. 50/mg).  The  National  Pituitary 
Agency  produces  650,000  international  units  (lU) 
(about  325  g)  of  hGH,  along  with  the  thyroid-stimulat- 
ing hormone,  prolactin,  and  other  hormones,  from 
about  60,000  human  pituitaries  collected  each  year. 
That  is  enough  hGH  both  for  the  current  demand  and 
for  perhaps  another  100  hypopituitary  patients. 

4.  Workers  at  the  Howard  Hughes  Medical  Institute  of 
the  University  of  California,  San  Francisco,  isolated 
messenger  RNA  from  a human  pituitary  tumor  and 
converted  it  into  a DNA-sequence  that  could  be  put  into 
E.  coli.  The  sequence,  however,  was  a mixture  of  hGH 
and  non-hGH  material.  It  has  been  reported  that  Eli  Lil- 
ly &,  Co.,  which  has  provided  some  grant  money  to  the 
Institute,  has  obtained  a license  to  the  patents  relating 
to  this  work.  Grants  from  the  National  Institutes  of 
Health  and  the  National  Science  Foundation  were  also 
acknowledged  in  the  publication. 


At  practically  the  same  time,  researchers  at  Genen- 
tech,  in  conjunction  with  their  associates  at  City  of 
Hope  National  Medical  Center  disclosed  the  production 
of  an  hGH  analog.  This  was  the  first  time  that  a human 
polypeptide  was  directly  expressed  in  E.  coli  in  func- 
tional form.  The  work  was  supported  by  Kabi  Gen  .AB, 
and  Kabi  has  the  marketing  rights. 

The  level  of  hGH  production  reported  in  the  scientif- 
ic account  of  the  Genentech  work  was  on  the  same 
order  as  that  reported  for  the  insulin  fragments— 
approximately  186,000  hGH  moUu'ules  per  c(’ll— a k‘\  ('l 
that  might  be  competitiv  e even  lud'orc*  efforts  are  made 
to  increase  yield.  Genentech  stresses  llu?  point  that  de- 
sign, rather  than  classical  mutation  and  selection,  is  the 
logical  way  to  improve  the  system,  since  the  hormotie's 
"blueprint"  is  incorporatcKl  in  a plasmid  that  can  he 
moved  between  strains  of  E.  coli,  betwc'en  s|)ecies,  oi' 
even  from  simple  bacteria  into  more  complex  orga- 
nisms, such  as  yeast. 

5.  Since  erythropoietin  is  a glyco|)rot(!in.  it  may  not  be 
feasible  to  synthesize  the;  active;  hor-mone  w ith  |)resent- 
ly  available  rDNA  techtii(|U(;s. 

6.  Antigens  are  surface  compoiuMits  o(  pathogenic  oiga- 
nisms,  toxins,  or  other  proteins  se'ci'cted  by  |)alhogenic 
micro-organisms.  I'Ikw  are;  also  the  specific  counter- 
parts of  antibodies:  antibodie^s  ar<‘  formed  by  the 
body’s  immune  .system  in  respotise*  to  their  presence 
Antibodies  are  synthesizeul  by  u bite  blood  cells  and  are 
created  in  such  a way  that  they  ai-e  uni(|uely  struc- 
tured to  bind  to  s|)ccific  antigens. 

7.  Many  of  the  most  d(;vastating  infectious  diseases  in- 
volve complex  parasit(!s  that  I'efuse  to  grow  under  lab- 
oratory conditions,  rlu;  first  cultivation  of  the  most 
malignant  of  the  s|)(;ci(;s  of  jiiotozoa  that  causes  ma- 
laria, using  human  ixul  blood  cells,  was  described  in 
1976  by  a Rockelell(;r  Univei  sity  jiarasitologist.  W ilium 
Rager,  Expm'iiiHintal  immunogens  were  prepared  and 
showed  [jromise  in  monkews,  but  concern  about  the  ex- 
istence of  the  r(;d  blood  ci>ll  remnants— w Inch  could 
give  rise  to  autoimmune  I'cactioiis— curtailed  the  pros 
pect  for  making  practical  vaccines  by  that  route  Sever 
al  biotechnology'  firms  are  currentlv  Irving  to  svnihe 
size  malai'ia  antigi'iis  by  molecular  cloning  I his  ellort 
may  product;  tt;chnic;il  solutions  to  such  si  oiirges  as 


Ch.  4— The  Pharmaceutical  Industry  • 81 


schistosDiiiiasis  (hilliar/ia).  tilariasis  (oncln)ceiTiasis 
and  elephantiasis),  leshmaniasis.  hookworm,  amehie  in- 
teetions,  aiui  lr\  panosomiasis  (slt*eping  sickness  and 
Chagas  disease). 

« .Another  potential  use  of  antigens  is  suggested  hv  the 
e.xperimental  treatment  ol  stage  I lung  cancer  patients 
with  vaccines  prepared  Irom  purified  human  lung 
cancer  antigens,  which  apjjears  to  suhstantially  pro- 
long survival.  And  the  Salk  Institute  is  expanding  clin- 
ical trials  in  vv  Inch  a pi’ocine  myelin  protein  prepared 
by  Kli  l.illv  <St  C'o.  is  injected  into  multiple  scleiosis  pa- 
tients to  mop  up  the  antimvelin  antibodies  that  those 
patients  are  jn-oducing.  Fifteen  to  forty-two  g of  myelin 
have  htHMi  injected  w ithout  adv  erse  effects,  suggesting 
a new  therapeutic  approach  to  auto  immune  diseases, 
rhe  protein  appears  to  sup()ress  the  sv  niptoms  of  ex- 
perimental allergic  encephalomyelitis,  an  animal  dis- 
ease resembling  multiple  sclerosis.  Should  this  re- 
search succeed,  the  use  of  molecular  clones  to  produce 
human  protein  antigens  seems  inev  itahle. 

9.  rhere  are  at  least  two  distinct  kinds  of  "classicar  inter- 
ferons—leukocyte  interferon  and  fibroblast  interferon, 
so-called  for  the  types  of  cells  from  which  they  are  ob- 
tained. A third  kind,  called  Iv  iiijthohlastoid  because  it  is 
produced  from  cells  deriv  ed  from  a Burkitt's  Iv  nipho- 
ma.  appears  to  be  a mixture  of  the  other  two  inter- 
ferons. All  produce  the  antiv  iral  state  and  are  induced 
by  viruses.  A fourth  kind,  known  as  "immune"  inter- 
feron, is  produced  by  Ivmphocytes.  Some  ev  idence  in- 
dicates that  it  may  he  a more  potent  antitumor  agent 
than  the  classical  types.  Currently,  interferon  is  ob- 
tained chiefly  from  white  blood  cells  (leukocytes)  from 
the  blood  bank  in  Helsinki  that  serv  es  all  of  Finland,  or 
from  fibroblasts  grown  in  cell  culture. 

10.  Recently,  G.  D.  Searle  & Co.  announced  that  new  tech- 
nologv"  developed  at  its  R&.D  facility  in  England  has  in- 
creased the  yield  of  fibroblast  interferon  by  a factor  of 
60.  On  tbe  basis  of  this  process,  Searle  expects  to  sup- 
ply material  for  the  first  large-scale  clinical  trial  of 
fibroblast  interferon.  Abbott  Laboratories  also  recently 
announced  plans  to  resume  production  of  limited 
quantities  of  fibroblast  interferon  for  clinical  studies  it 
plans  to  sponsor. 

L'nlike  leukocytes  and  specially  treated  fibroblasts, 
which  can  be  used  only  once,  lymphoblasts  derived 
from  the  tumor  Burkitt's  lymphoma  grow  freely  in 
suspension  and  produce  the  least  costly  interferon 
presently  obtainable.  However,  they  also  produce  a dis- 
advantageous mixture  of  both  leukocyte  and  fibroblast 
interferons.  The  Burroughs-Wellcome  Co.  produces 
lymphoblastoid  interferon  in  1,000-1  fermenters  and 
has  begun  clinical  trials  in  England,  but  the  U.S.  FD,A 
has  generally  resisted  efforts  to  make  use  of  products 
derived  from  malignant  cells.  It  is  used  extensively  in 
research,  and  FDA  is  considering  evidence  from  Bur- 
roughs-W'ellcome  that  may  lead  to  a relaxation  of  the 
prohibition,  under  pressure  from  the  National  Cancer 
Institute. 

11.  What  may  be  a landmark  patent  has  been  issued  to 
Hilary  Koprowski  and  Carlos  Croce  of  the  Wistar  Insti- 


tute (for  work  done  under  the  then  Department  of 
Health,  Education,  and  Welfare  funding)  on  the  pro- 
duction of  monoclonal  antibodies  against  tumor  cells. 
In  a number  of  examples,  these  reseaichers  demon- 
strated that  an  animal  can  be  immunized  with  tumor 
cells,  and  that  hyhridomas  derived  from  that  animal 
will  produce  antihodies  that  demonstrate  a specificity 
for  the  tumor. 

rhe  final  sentence  of  the  patent  text  provides  the  ra- 
tionale for  the  use  of  antibodies  in  both  cancer  and  in- 
fectious disease  therapies:  "If  the  (tumor)  antigen  is 
present,  the  patient  can  be  given  an  injection  of  an  anti- 
body as  an  aid  to  react  with  the  antigen."  (II. S. 
4,172,124.) 

12.  .Myeloma  cells  grow  v igorously  in  culture  and  have  the 
uniciue  chaiacteristic  of  producing  large  quantities  of 
antihodi(!s.  Each  spleen  cell  of  the  immune  type,  on  the 
other  hand,  produces  an  antibody  that  recognizes  a 
single  antigen,  hut  these  do  not  grow  well  in  culture. 
W hen  normal  immune  spleen  cells  are  fused  with  mye- 
loma cells,  the  resulting  mixture  of  genetic  capacities 
forms  a cell,  called  a "hybridoma, " which  displays  the 
desired  characteristics  of  the  parent  cells:  1)  it  secretes 
the  antibody  specified  by  the  genes  of  the  spleen  cell; 
and  2)  it  disjjlays  the  v igorous  grow  th,  production,  and 
longev  ity  that  is  typical  of  the  myeloma  cell. 

13.  rhe  use  of  high-correlation  antibody  assays  in  cancer 
studies  bas  only  just  begun.  Antibodies  that  have  been 
treated  so  they  can  be  seen  with  X-rays  and  that  are 
specific  for  a tumor,  can  be  used  early  to  detect  the  oc- 
currence or  spread  of  tumor  cells  in  the  body.  Because 
some  785,000  new  cancer  cases  will  be  detected  in 
1980  with  current  diagnostic  methods,  because  cancer 
will  cause  405,000  deaths,  and  because  early  detection 
is  the  major  key  to  improving  survival,  the  implications 
are  indeed  enormous. 

14.  In  the  late  1950’s,  Lederle  Laboratories  marketed  a 
preparation  of  95-percent  pure  streptokinase  (a  bac- 
terially  produced  enzyme  that  dissolves  blood  clots)  for 
intravenous  administration.  They  withdrew  the  prod- 
uct from  the  market  around  1960  because  it  caused 
allergic  reactions,  which  dampened  clinical  enthusiasm 
for  its  therapeutic  potential. 

The  presence  in  human  urine  of  urokinase,  an  en- 
zyme also  capable  of  removing  blood  clots,  was  also  dis- 
covered in  the  early  1950's.  Urokinase  was  purified, 
crystallized,  and  brought  into  clinical  use  in  the  mid- 
1960’s.  From  the  beginning  it  was  apparent  that  “an  in- 
tense thrombolytic  state  could  be  achieved  with  a 
much  milder  coagulation  defect  than  occurred  with 
streptokinase;  no  pyrogenic  or  allergic  reactions  were 
noted,  and  no  antibodies  resulted  from  its  administra- 
tion . . , There  did  not  appear  to  be  as  great  variation  in 
patient  responsiveness.”  In  1967-68  and  1970-73,  the 
National  Heart  and  Lung  Institute  organized  clinical 
trials  that  compared  urokinase  with  streptokinase  and 
heparin,  an  anticoagulant,  in  the  treatment  of  pul- 
monary embolism.  The  trials  indicated  that  strep- 
tokinase and  urokinase  were  equivalent  and  superior 
to  heparin  over  the  short  term,  although  their  long- 


82  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


term  benefits  were  not  established.  Since  then,  clinical 
investigation  of  urokinase  has  been  hampered  by 
domestic  regulatory  problems,  which  have  raised  the 
cost  of  production  and  restricted  its  availability  in  the 
United  States. 

In  January  1978,  Abbott  Laboratories  obtained  a 
new  drug  application  for  urokinase  and  introduced  the 
product  Abbokinase;  by  that  time,  however,  the  sales 
of  urokinase  in  Japan  were  already  pushing  $90  million 
per  year.  Recently,  Sterling  Drug  has  begun  marketing 
a urokinase  product  (Breokinase)  manufactured  by 
Green  Cross  of  Japan:  "According  to  Japanese  reports, 
urokinase  is  the  first  Japanese-made  drug  formulation 
to  receive  production  and  sales  approval  from  FDA. 
Green  Cross  estimates  that  within  3 years  of  the  start 
of  Sterling’s  marketing  activities,  the  value  of  uro- 
kinase exports  will  reach  Yen  500  million  ($2.12  mil- 
lion) per  month,  and  considers  that  its  profits  from  ex- 
porting a finished  product  will  probably  be  better  than 
those  from  bulk  drug  sales  or  the  licensing  of  technol- 
ogy.’’ The  Green  Cross  product  is  made  from  human 
urine  collected  throughout  Korea  and  Japan,  and  takes 
advantage  of  technology  licensed  from  Sterling.  Ab- 
bott’s product,  on  the  other  hand,  is  derived  from 
kidney-cell  culture. 


15.  Intergeneric  hybrids  have  extremely  interesting  pos- 
sibilities. For  example,  it  would  be  beneficial  to  cepha- 
losporin-process technology  to  combine  in  one  orga- 
nism the  acyltransferase  from  Penicillium  chrysogenum 
and  the  enzymes  of  C.  acremonium,  which  does  not  in- 
corporate side  chain  precursors  onto  cephalosporin 
like  P.  chrysogenum  does  for  penicillins. 

16.  Another  example  of  recombination  between  species  is 
that  reported  for  two  species  of  fungi,  Aspergillus  nidu- 
lans  and  A.  rugulosus,  subsequent  to  protoplast  fusion. 

The  only  report  of  a successful  cross  between 
genera  using  protoplast  fusion  technology  has  been  be- 
tween the  yeasts  Candida  tropicalis  and  Saccario- 
mycopsis  fibuligera,  which  took  place  at  low  frequency 
and  gave  rise  to  types  intermediate  between  the 
parents. 

17.  An  example  of  screening  is  provided  by  the  new 
i8-lactam  (penicillin-like)  antibiotics.  Using  older 
screening  methods,  no  new  ;8-lactams  were  found 
from  1956  until  1972  when  a new  method  was  devised. 
A new  series  of  these  antibiotics  was  thus  found. 
Within  the  past  year,  6 new  )3-lactams  have  been 
commercialized  and  at  least  12  more  are  in  clinical 
trials  around  the  world.  The  sales  forecasts  for  these 
new  agents  are  estimated  to  he  o\  er  $1  billion. 


chapter  5 


Page 


Background 85 

Overview  of  the  Industry 85 

Fermentation  and  the  Chemical  Industry 87 

New  Process  Introduction 89 

Characteristics  of  Biological  Production 

Technologies 90 

Renewable  Resources 90 

Physically  Milder  Conditions 91 

One-Step  Production  Methods 91 

Reduced  Pollution 91 

Industrial  Chemicals  That  May  Be  Produced  by 

Biological  Technologies 92 

Fertilizers,  Polymers,  and  Pesticides 94 

Constraints  on  Biological  Production  Techniques  96 

An  Overview  of  Impacts 97 

Impacts  on  Other  Industries 97 

Impacts  on  University  Research 98 

The  Social  Impacts  of  Local  Industrial  Activity  . 99 
Impacts  on  Manpower 99 


Tables 


Table  No.  Page 

9.  Data  for  Commercially  Produced  Amino 

Acids 88 


10.  Summary  of  Recent  Estimates  of  Primary 
U.S.  Cost  Factors  in  the  Production  of  L- 
Lysine  Monohydrochloride  by  Fermentation 


and  Chemical  Synthesis 89 

11.  Some  Commercial  Enzymes  and  Their  Uses.  . 93 

12.  Expansion  of  Fermentation  Into  the 

Chemical  Industry 94 

13.  The  Potential  of  Some  Major  Polymeric 
Materials  for  Production  Using  Biotechnology  95 

14.  Some  Private  Companies  With 

Biotechnology  Programs 98 

15.  Distribution  of  Applied  Genetics  Activity 

in  Industry 100 

16.  Manpower  Distribution  of  a Firm  With 

Applied  Genetics  Activity 100 

17.  Index  to  Fermentation  Companies 100 

18.  Fermentation  Products  and  Producers 101 

19.  U.S.  Fermentation  Companies 103 

Figures 

Figure  No.  Page 

24.  Flow  of  Industrial  Organic  Chemicals  From 

Raw  Materials  to  Consumption 86 

25.  Diagram  of  Alternative  Routes  to  Organic 

Chemicals 90 


chapter  5 

The  Chemical  Industry 


Background  

The  organic  substances  first  used  hv  humans 
to  make  useful  materials  such  as  cotton,  linen, 
silk,  leather,  adhesives,  and  dyes  were  obtained 
from  plants  and  animals  and  are  natural  and  re- 
newable resources.  In  the  late  19th  century, 
coal  tar,  a tionrenewahle  substance,  was  found 
to  he  an  e.xcellent  raw  material  for  many  organ- 
ic compounds.  When  organic  chemistry  devel- 
oped as  a science,  chemical  technologv'  im- 
proved. .At  about  the  same  time  relatively  cheap 
petroleum  became  vv  idely  av  ailahle.  The  indus- 
try shifted  rapidly  to  using  petroleum  as  its  ma- 
jor raw  material. 

The  chemical  industry's  constant  search  for 
cheap  and  plentiful  raw  materials  is  now  about 
to  come  full  circle.  The  supply  of  petroleum, 
which  presently  serv  es  more  than  90  percent  of 
the  industrv’s  needs,  is  severely  threatened  by 
both  dw  indling  resources  and  increased  costs.  It 
has  been  estimated  that  at  the  current  rate  of 
consumption,  the  world's  petroleum  supplies 
w ill  be  depleted  in  the  middle  of  the  ne.xt  cen- 
tury. Most  chemical  industry  analysts,  there- 
fore, foresee  a shift  first  back  to  coal  and  then, 
once  again,  to  the  natural  renewable  resources 
referred  to  as  biomass.  The  shifts  will  not 
necessarily  occur  sequentially  for  the  entire 


Overview  of  the  industry 

The  chemical  industry  is  one  of  the  largest 
and  most  important  in  the  world  today.  The  U.S. 
market  for  synthetic  organic  chemicals  alone, 
e.xcluding  primary  products  made  from  petro- 
leum, natural  gas,  and  coal  tar,  exceeded  S35 
billion  in  1978. 

The  industry's  basic  function  is  to  transform 
low-cost  raw  materials  into  end-use  products  of 
greater  value.  Tbe  most  important  raw  materi- 
als are  petroleum,  coal,  minerals  (pbospbate, 
carbonate),  and  air  (oxygen,  nitrogen).  Roughly 
two-thirds  of  the  industry  is  devoted  to  produc- 


chemical industry.  Rather,  both  coal  and  bio- 
mass will  be  examined  for  tbeir  potential  roles 
on  a product-by-product  basis.' 

Tbe  chemical  industry  is  familiar  with  the 
technology  of  converting  coal  to  organic  chem- 
icals, and  a readily  available  supply  exists.  Coal- 
based  technologies  will  he  used  to  produce  a 
w ide  arrav  of  organic  chemicals  in  the  near  fu- 
ture.* Nevertheless,  economic,  env  ironmental, 
and  technical  factors  will  increase  the  industry’s 
intei'est  in  biomass  as  an  alternativ  e source  tor 
raw  materials.  .Applied  genetics  will  probably 
plav  a major  role  in  enhancing  the  possibilities 
l)v  allowing  biomass  and  carbohydrates  from 
natural  sources  to  be  converted  into  various 
chemicals.  Biology  will  thereby  take  on  the  dual 
role  of  prov  iding  both  raw  materials  and  a proc- 
ess for  production. 


'For  I'lii’lhpi'  dclails  see  Energy  From  Biological  Processes,  \ol.  I, 
or.V-K-124  (W  ashington,  O.C.:  OtI'iee  ot  Technology  Assessment, 
July  19801. 

VVIost  important  organic  intermediates  (chemical  compounds 
used  lor  the  industrial  synthesis  of  commercial  products  such  as 
plastics  and  fihers)  can  be  obtained  from  coal  as  an  alternative  raw 
material.  Currently,  methods  are  being  dexeloped  to  convert  coal 
into  "synthetic  gas,"  which  can  then  be  used  as  raw  material  for 
further  conversions. 


ing  inorganic  chemicals  such  as  lime,  salt,  am- 
monia, carbon  dioxide,  chlorine  gas,  and  hydro- 
choloric  and  other  acids. 

The  other  third,  which  is  the  target  for  bio- 
technology, produces  organic  chemicals.  Its  out- 
put includes  plastics,  synthetic  fibers,  organic 
solvents,  and  synthetic  rubber.  (See  figure  24.) 
In  general,  petroleum  and  natural  gas  are  first 
converted  into  “primary  products”  or  basic  or- 
ganic chemicals  such  as  the  hydrocarbons  ethyl- 
ene and  benzene.  These  are  then  converted  into 
a wide  range  of  industrial  chemicals.  Ethylene 


85 


86  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animais 


Figure  24.— Flow  of  Industrial  Organic  Chemicals  From  Raw  Materials  to  Consumption 


Organic  resources 

80%  raw  material  from  petroleum/ 
natural  gas 


20%  raw  material  from  coal,  coke,  and 
renewable  resources 


SOURCE:  U.s.  Industrial  Outlook  (Washington.  D C.:  Department  of  Commerce,  1978);  Kline  Guide  to  Chemical  Industry.  Fairfield. 
N.J.,  adapted  from  Tong,  1979. 


Ch.  5 — The  Chemical  Industry  • 87 


alone  ser\  es  as  the  basic  chemical  tor  the  manu- 
facture of  half  of  the  largest  \olume  industi'ial 
chemicals.  Kach  of  the  steps  in  a chemical  con- 
\ ersion  process  is  controlled  hv  a separate  reac- 
tion, u hich  is  often  performed  hv  a separate 
compan\-. 

pAaluating  the  competiti\eness  both  of  a 
process  and  of  the  market  is  critical  for  the 
chemical  industry,  which  is  intensixe  for  cap- 
ital, energy,  and  raw  materials.  Its  plants  use 
large  amounts  of  energx’  atid  can  cost  hundreds 
of  millions  of  dollars  to  build,  and  raw  material 
costs  are  generally  5t)  to  80  percent  of  a prod- 
uct’s cost.  If  a biological  process  can  use  the 
same  raw  materials  and  reduce  the  process  cost 
by  even  20  percent,  or  allow  the  use  of  inexpen- 
sive raw  materials,  it  could  prox  ide  the  industry 
xvith  a major  price  break. 

Fermentation  and 
the  chemical  industry 

The  production  of  industrial  chemicals  by 
fermentation  is  not  nexv.  Scores  of  chemicals 
hax  e been  produced  by  micro-organisms  in  the 
past,  only  to  be  replaced  by  chemical  produc- 
tion based  on  petroleum.  In  1946,  for  example, 
27  percent  of  the  ethyl  alcohol  in  the  United 
States  xvas  produced  from  grain  and  grain  prod- 
ucts, 27  percent  from  molasses,  a fexv  percent 
each  from  such  materials  such  as  potatoes,  pine- 
apple juice,  cellulose  pulp,  and  xvhey,  and  only 
36  percent  from  petroleum.  Ten  years  later 
almost  60  percent  xvas  derived  from  petroleum. 

Exen  more  dramatically,  fumaric  acid  xvas  at 
one  time  produced  on  a commercial  scale 
through  fermentation,  but  its  biological  produc- 
tion xvas  stopped  xvhen  a more  economical  syn- 
thesis from  benzene  xvas  dex  eloped.  Frequently, 
after  a fermentation  product  xvas  discovered, 
alternative  chemical  synthetic  methods  xvere 
soon  dexeloped  that  used  inexpensive  petro- 
leum as  the  raxv  niaterial. 

Nevertheless,  for  the  fexv  chemical  entities 
still  produced  by  fermentation,  applied  genetics 


has  contributed  to  the  economic  viability  of  the 
process.  The  production  of  citric  and  lactic 
acids  and  xarious  amino  acids  are  among  the 
processes  that  haxe  benefited  from  genetics. 
Lactic  acid  is  produced  both  synthetically  and 
by  fei'inentation.  t)x  er  the  past  10  to  20  years, 
manufacture  by  fermentation  has  experienced 
competition  from  chemical  processes. 

The  organisms  used  for  the  production  of  lac- 
tic acid  are  x arious  species  of  the  bacterium  Lac- 
tobacillus. Starting  materials  may  be  glucose,  su- 
crose, or  lactose  (xvhey).  The  fermentation  per 
se  is  efficient,  I'esulting  in  90  percent  yields,  de- 
pending on  the  original  carbohydrate.  Since 
most  of  the  problems  in  the  manufacture  of  lac- 
tic acid  lie  in  the  recox  ery  procedure  and  not  in 
fermentation,  fexx’  attempts  have  been  made  to 
improxe  the  industrial  processes  through 
genetics. 

Citric  acid  is  the  most  important  acidulant, 
and  historically  has  held  oxer  55  to  65  percent 
of  the  acidulant  market  for  foods.*  It  is  also 
used  in  pharmaceuticals  and  miscellaneous  in- 
dustrial applications.  It  is  produced  commercial- 
ly by  the  mold  Aspergillus  niger.  Surprisingly  lit- 
tle xvork  has  been  published  on  improving  citric 
acid-producing  strains  of  this  micro-organism. 
W eight  yields  of  110  percent  have  recently  been 
reported  in  A.  niger  mutants  obtained  by  ir- 
radiating a strain  for  which  a maximum  yield  of 
29  percent  had  been  reported. 

Amino  acids  are  the  building  blocks  of  pro- 
teins. Txxenty  of  them  are  incorporated  into 
proteins  manufactured  in  cells,  others  serve 
specialized  structural  roles,  are  important  meta- 
bolic intermediates,  or  are  hormones  and  neu- 
rotransmitters. All  of  the  amino  acids  are  used 
in  research  and  in  nutritional  preparations, 
xvith  most  being  used  in  the  preparation  of 
pharmaceuticals.  Three  are  used  in  large  quan- 
tities for  txvo  purposes:  glutamic  acid  to  manu- 
facture monosodium  glutamate,  which  is  a fla- 


*The  other  two  important  acidulants,  or  acidifying  agents,  are 
phosphoric  acid  (20  to  25  percent)  and  malic  acid  (5  percent). 


88  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


vor  enhancer  particularly  in  oriental  cooking;* 
and  lysine  and  methionine  as  animal  feed  ad- 
ditives. 

Conventional  technology  for  producing  glu- 
tamic acid  is  based  on  pioneering  work  that  was 
subsequently  applied  to  other  amino  acids.  The 
production  employed  microbial  strains  to  pro- 
duce amino  acids  that  are  not  within  their  nor- 
mal biosynthetic  capabilities.  This  was  accom- 


*Monosodium  glutamate  is  the  sodium  salt  of  glutamic  acid.  In  1978, 
about  18,000  tonnes  were  manufactured  in  the  United  States  and  about 
11,000  tonnes  imported.  The  food  industy  consumed  97  percent.  The 
fermentation  plant  of  the  Stauffer  Chemical  Co.  in  San  Jose,  Calif.,  is 
the  sole  U.S.  producer.  The  microbes  used  in  glutamic  acid  fermenta- 
tion [Corynebacterium  glutamicum,  C.  lileum,  and  Brevibacterium 
flavum)  produce  it  in  60  percent  of  theoretical  yield.  Thus,  there  is  some 
but  not  great  potential  for  the  use  of  applied  genetics  to  improve  the 
yield.  Many  of  the  genetic  approaches  have  already  been  thoroughly 
investigated  by  industrial  scientists. 


plished  by  using  two  methods:  1)  manipulating 
microbial  growth  conditions,  and  2)  isolating 
naturally  occurring  mutants. 

Although  microbial  production  of  all  the 
amino  acids  has  been  studied,  glutamic  acid  and 
L-lysine**  are  the  ones  produced  in  significant 
quantities  by  fermentation  processes.  (See  table 
9.)  The  production  of  L-lysine  is  an  e.xcellent  e.\- 

**The  lack  of  a single  amino  acid  can  retard  protein  synthesis,  and 
therefore  growth,  in  a mammal.  The  limiting  amino  acid  is  a function  of 
the  animal  and  its  feed.  The  major  source  of  animal  feed  in  the  United 
States  is  soybean  meal.  The  limiting  amino  acid  for  feeding  swine  is 
methionine;  the  limiting  amino  acid  for  feeding  poultry  is  lysine. 
Because  of  increased  poultry  demand,  world  demand  for  lysine  is 
climbing.  Eurolysine  is  spending  $27  million  to  double  its  production 
capacity  in  Amiens,  France,  to  10  thousand  tonnes.  The  Asian  and 
Mideast  markets  are  estimated  to  increase  to  3 thousand  tonnes  in 
1985.  Some  bacteria  produce  lysine  at  over  90  percent  of  theoretical 
yield.  Little  genetic  improvement  is  likely  in  this  conversion  yield, 
however,  significant  improvement  can  be  made  in  the  rate  and  final 
concentration. 


Table  9.— Data  for  Commercially  Produced  Amino  Acids^ 


Price  March 

Potential  for  application  of 

1980 (per  kg 

Production  1978 

biotechnology  (de  novo  synthesis  or 

Amino  acid 

pure  L) 

Present  source 

(tonnes) 

bioconversion;  organisms  and  enzymes) 

Alanine 

$ 80 

Hydrolysis  of  protein; 

10-50(J)b 

— 

chemical  synthesis 

Arginine 

28 

Gelatin  hydrolysis 

200  - 300  (J) 

Fermentation  in  Japan 

Asparagine 

50 

Extraction 

10-50  (J) 

— 

Aspartic  acid 

12 

Bioconversion  of 

fumaric  acid 

500-1,000  (J) 

Bioconversion 

Citrulline 

250 

— 

10-90(J) 

Fermentation  in  Japan 

Cysteine 

50 

Extraction 

100-200  (J) 

— 

Cystine 

60 

Extraction 

100-200  (J) 

— 

DOPA  (dihydrophenylalanine)  . 750 

Chemical 

100-200  (J) 

— 

Glutamic 

4 

Fermentation 

10,000-100,000  (J) 

De  novo:  Micrococcus  glutamicus 

Glutamine 

55 

Extraction 

200  - 300  (J) 

Fermentation  in  Japan 

Histidine 

160 

— 

100-200 

Fermentation  in  Japan 

Hydroxyproline 

280 

Extraction  from  collagen  10  - 50 

— 

Isoleucine 

350 

Extraction 

10-50  (J) 

— 

Leucine 

55 

— 

50-100  (J) 

Fermentation  in  Japan 

Lysine 

350 

Fermentation  (80%) 

10,000  (J) 

(80%  by  fermentation)  De  novo: 

Chemical  (20%) 

Corynebacterium  glutamicum  and 
Brevibacterium  tlavum 

Methionine 

265 

Chemical  from  acrolein 

17.000  (D,L)c 

20.000  (D,L)  (J) 

— 

Ornithine 

60 

— 

10-50  (J) 

Fermentation  in  Japan 

Phenylalanine 

55 

Chemical  from 

50-100  (J) 

Fermentation  in  Japan 

benzaldehyde 

Proline 

125 

Hydrolysis  of  gelatin 

10-50  (J) 

Fermentation  in  Japan 

Serine 

320 

— 

10-50  (J) 

Bioconversion  in  Japan 

Threonine 

150 

— 

50-10(J) 

Fermentation  in  Japan 

Tryptophan 

110 

Chemical  from  indole 

55  (J) 

— 

Tyrosine 

13 

Extraction 

50-100  (J) 

-- 

Valine 

60 

— 

50-100  (J) 

Fermentation  in  Japan 

^Production  data  largely  from  Japan  because  of  relative  small  U.S.  production. 
*^Japan. 

'-D  and  L forms. 

SOURCE:  Massachusetts  Institute  of  Technology. 


Ch.5 — The  Chemical  Industry  • 89 


amjile  of  the  eoiniietition  hetween  ehemieal  and 
hioleehnologieal  methods.  I'ermentation  lias 
been  gradually  r(>plaeing  its  produetion  In 
ehemieal  s\tithesis:  in  td.SO,  80  peieenl  of  its 
worldu  ide  produetion  is  e.vpeeled  to  ht>  In  mi- 
crobes. It  is  not  produced  in  the  Ihiited  States, 
which  imported  about  7,000  tonnes  in  1979, 


mostly  from  Japan  and  South  Korea.  Recent 
estimates  of  primary  U.S.  cost  factors  in  the 
competing  production  methods  are  summarized 
in  table  10.  Fermentation  costs  are  lower  for  all 
three  components  of  direct  operating  costs; 
labor,  material,  and  utilities. 


Table  10.— Summary  of  Recent  Estimates  of  Primary  U.S.  Cost  Factors  in  the  Production  of 
L-Lysine  Monohydrochloride  by  Fermentation  and  Chemical  Synthesis 


Cost  factors  in  production  of  98%  L-lysine  monohydrochloride 


By  fermentation^ 

By  chemical  synthesis^’ 

Requirement 
(units  per  unit 

Estimated  1976  cost 
per  unit  product 

Requirement 
(units  per  unit 

Estimated  1976  cost 
per  unit  product 

product) 

Cents/lb  Cents/kg 

product) 

Cents/lb  Cents/lb 

Total  laborF 

— 

8 

18 

— 

9 

20 

Materials 

Molasses 

44 

7 

16 

— 

— 

— 

Soybeanmeal,  hydrolized  . . . 

0.462 

4 

9 

— 

— 

— 

Cyclohexanol 

— 

— 

— 

0.595 

17 

37 

Anhydrous  ammonia 

— 

— 

— 

0.645 

6 

14 

Other  chemicals'^ 

— 

7 

15 

— 

4 

10 

Nutrients  and  solvents 

— 

— 

— 

— 

4 

8 

Packaging,  operating,  and 

maintenance  materials . . . 

— 

10 

22 

— 

9 

21 

Total  materials 

— 

28 

62 

— 

45 

90 

Total  utilities^ 

— 

6 

12 

— 

7 

16 

Total  direct  operating  cost 

— 

42 

92 

— 

56 

126 

Plant  overhead,  taxes. 

and  insurance 

— 

10 

21 

— 

10 

21 

Total  cash  cost 

— 

52 

11 

— 

66 

147 

Depreciation* 

— 

16 

35 

— 

13 

28 

Interest  on  working  capital 

— 

1 

3 

— 

1 

3 

Total  cost9 

— 

69 

151 

— 

80 

178 

^Assumes  a 23-percent  yield  on  molasses. 
t>Assumes  a 65-percent  yield  on  cyclohexanol. 

'-Includes  operating,  maintenance,  and  control  laboratory  labor. 

•tpor  both  the  process  of  fermentation  and  chemical  synthesis,  assumed  use  of  hydrochloric  acid  (36  percent)  and  ammonia  (29  percent).  For  fermentation  includes  also 
potassium  diphosphate,  urea,  ammonium  sulfate,  calcium  carbonate,  and  magnesium  sulfate.  For  chemical  synthesis  also  includes  nitrosyl  chloride,  sulfuric  acid, 
and  a credit  for  ammonium  sulfate  byproduct. 

®Total  utilities  for  both  processes  include  cooling  water,  steam  process  water,  and  electricity.  For  chemical  synthesis,  natural  gas  is  also  included. 

*Ten  percent  per  year  of  fixed  capital  costs  for  a new  20  million  lb  per  year  U.S.  plant  built  in  1975  at  assumed  capital  cost  of  $38.6  x 10‘  for  fermentation  and  $32.5  x 10“ 
for  chemical  synthesis  exclusive  of  land  costs. 

SOURCE:  Stanford  Research  Institute.  Chemical  Economics  Handbook  583:3401,  May  1979. 


New  process  introduction 

The  development  of  biotechnology  should  be 
viewed  not  so  much  as  the  creation  of  a new  in- 
dustry as  the  rex’italization  of  an  old  one.  Both 
fermentation  and  enzyme  technologies  will 
have  an  impact  on  chemical  process  de\  elop- 
ment.  The  first  will  affect  the  transition  from 
nonrenewable  to  renewable  raw  materials.  The 


second  will  allow  fermentation-derived  prod- 
ucts to  enter  the  chemical  conversion  chains, 
and  will  compete  directly  with  traditional  chem- 
ical transformations.  (See  figure  25.)  Fermenta- 
tion, by  replacing  various  production  steps, 
could  act  as  a complementary  technology  in  the 
overall  manufacture  of  a chemical. 


90  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Figure  25.— Diagram  of  Alternative  Routes  to  Organic  Chemicals 


®C  followed  by  number  Indicates  length  of  carbon  chain. 

SOURCE:  G.  E.  Tong,  "Industrial  Chemicals  From  Fermentation  Enzymes,"  Microb.  Techno!.,  vol.  1, 1979,  pp.  173-179. 


Characteristics  of  biological 
production  technologies 

The  major  advantages  of  using  commercial 
fermentation  include  the  use  of  renewable  re- 
sources, the  need  for  less  extreme  conditions 
during  conversion,  the  use  of  one-step  produc- 
tion processes,  and  a reduction  in  pollution.  A 
micro-organism  might  he  constructed,  for  ex- 
ample, to  transform  the  cellulose  in  wood  di- 
rectly into  ethanol.*  (App.  I-D,  a case  study  of 
the  impact  of  genetics  on  ethanol  production, 
elaborates  these  points.) 

RENEWABLE  RESOURCES 
Green  plants  use  the  energy  captured  from 
sunlight  to  transform  carbon  dioxide  from  the 

*A  retiiie.st  for  approval  ol  suoli  an  aocompILshnient  liv  rONA 
tcchni(|ue.s  was  siihmittecl  to  llie  Heoomliinanl  ONA  Achisorv 
Committee  at  the  Sept.  25,  1980  meeting. 


atmosphere  into  carbohydrates,  some  of  \\  hi('h 
are  used  for  their  own  energy  ikmhIs.  Tlu'  rest 
are  accumulated  in  starches,  cellulose',  lignins, 
and  other  materials  called  the  biomass,  which  is 
the  foundation  of  all  renewable  resources. 

The  technologies  of  genetic  (’iiginee'i  ing  could 
help  ease  the  chemical  industry's  dependence' 
on  petroleum-based  products  by  making  the'  use' 
of  renewable  resources  attractive'.  .\ll  mie  re)- 
organisms  can  metaheilize  e'arhohyelrate's  anel 
convert  them  to  various  end  proelucts.  Ivxte'n- 
sive  research  and  devele)pment  (H&.D)  has 
already  been  conducted  on  thei  pe)ssihility  ol 
using  genetically  engineei'ed  strains  te>  ceiine'i  t 
cellulose,  the  major  carbohydrate'  in  plants,  te> 
commercial  products.  I he  basic  huileiing  hleie  k 
of  cellulose— glucose— can  he  re'aelily  use*d  as  .1 
raw  material  for  fermentatiein. 


c/7.5 — The  Chemical  Industry  • 91 


OtlK’r  plant  carbohydi'atos  include  corn- 
starch, molasses,  and  lignin.  I'he  last,  a polymer 
tbiind  in  wood,  could  he  used  as  a |)i'ecursor  I'oi' 
the  hiosynthesis  of  aromatic  (benzene-like) 
chemicals,  making  their  production  simpler  and 
moi’e  economical.  ,\e\ ertheless,  the  increase  in 
the  |)i'i('e  of  peti'oleum  is  not  a sufficient  reason 
for  switching  raw  matei'ials,  sinc'e  the  cost  of 
carholn cirates  and  other  biological  materials 
has  been  inci'easing  at  a relati\  e rate. 

PHVSICALI.V  MILDER  CONDITIO, \S 

In  general,  there  are  two  main  ways  to  speed 
chemical  I'eactions:  by  increasing  the  reaction 
tempei'ature  and  by  adding  a catalyst.  ,\  catalyst 
(usualK  a metal  oi'  metal  com[)le.\l  causes  one 
specific  reaction  to  occur  at  a faster  rate  than 
others  in  a chemical  mi.xtui’e  by  [)io\  iding  a sur- 
face on  which  that  reaction  can  he  pi'omoted. 
E\en  using  the  most  effecti\  e catalyst,  the  con- 
ditions needed  to  accelerate  industrial  organic 
reactions  often  require  e.xtremely  high  tem- 
peratures and  pressures— sexeral  hundred  de- 
grees Celsius  and  se\eral  hundred  pounds  per 
square  inch. 

Biological  catalysts,  or  enzymes,  on  the  other 
hand  can  speed-up  reactions  without  the  need 
for  such  e.xtreme  conditions.  Reactions  occur  in 
dilute,  aqueous  solutions  at  the  moderate  condi- 
tions of  temperature,  pressure,  and  pH  (a  meas- 
ure of  the  acidity  or  alkalinitx’  of  a solution)  that 
are  compatible  w ith  life. 

ONE-STEP  PRODUCTION  METHODS 

In  the  chemical  synthesis  of  compounds,  each 
reaction  must  take  place  separately.  Because 
most  chemical  reactions  do  not  yield  pure  prod- 
ucts, the  product  of  each  indi\  idual  reaction 
must  be  purified  before  it  can  be  used  in  the 
next  step.  This  approach  is  time-consuming  and 
expensixe.  If,  for  example,  a synthetic  scheme 
that  starts  with  ethylene  (a  petroleum-based 
product)  requires  10  steps,  with  each  step  yield- 
ing 90  percent  product  (very  optimistic  yields  in 
chemical  syntheses),  only  about  one-third  of  the 
ethylene  is  conx  erted  into  the  final  end  product. 
Purification  may  be  costly;  often,  the  chemicals 
inx’olx  ed  (such  as  organic  solx  ents  for  extrac- 
tions) and  the  byproducts  of  the  reaction  are 
toxic  and  require  special  disposal. 


In  biological  systems,  micro-organisms  often 
complete  entire  synthetic  schemes.  The  conver- 
sion takes  place  essentially  in  a single  step, 
although  sexeral  might  occur  within  the  orga- 
nisms, XX  hose  enzymes  can  transform  the  pre- 
cursor through  the  intermediates  to  the  desired 
end  product.  Purification  is  not  necessary. 

REDUCED  POLLliTION 

.Metal  catalysts  are  often  nonspecific  in  their 
action:  xxhile  they  may  promote  certain  reac- 
tions, their  actions  are  not  ordinarily  limited  to 
making  only  the  desired  products.  Consec|uent- 
ly,  they  haxe  sexei'al  undesirable  features:  the 
formation  of  side-products  or  byproducts;  the 
incomplete  conxersion  of  the  starting  materi- 
al(s);  and  the  mechanical  and  accidental  loss  of 
the  product. 

The  last  pi'ohlem  occurs  xvith  all  types  of  syn- 
thesis. rhe  first  txxo  represent  inefficiencies  in 
the  use  of  the  raxv  materials,  lliese  necessitate 
the  separation  and  recycling  of  the  side-prod- 
ucts formed,  xvhich  can  he  difficult  and  costly 
because  they  are  often  chemically  and  physi- 
cally similar  to  the  desired  end  products.  (Most 
separation  techniques  are  based  on  differences 
in  physical  properties— e.g.,  density,  volatility, 
and  size.) 

W hen  byproducts  and  side-products  have  no 
x alue,  or  xvhen  unconx  erted  raxv  material  can- 
not be  recycled  economically,  problems  of 
xxaste  disposal  and  pollution  arise.  Their  solu- 
tion requires  ingenuity,  xlgilance,  energy,  and 
dollars.  Many  present  chemical  processes  create 
useless  xxastes  that  require  elaborate  degrada- 
tion procedures  to  make  them  environmentally 
acceptable.  In  1980,  the  chemical  industry  is  ex- 
pected to  spend  S883  million  on  capital  outlays 
for  pollution  control,  and  xvell  over  $200  million 
on  R&D  for  new'  control  techniques  and  re- 
placement products.  These  figures  do  not  in- 
clude the  millions  of  dollars  that  have  been 
spent  in  recent  years  to  clean  up  toxic  chemical 
dumps  and  to  compensate  those  harmed  by 
poorly  disposed  xvastes,  nor  do  they  include  the 
cost  of  energy  and  labor  required  to  operate 
pollution-control  systems. 

A genetically  engineered  organism,  on  the 
other  hand,  is  designed  to  be  precursor-  and 


92  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


product-specific,  with  each  enzyme  having 
essentially  100-percent  conversion  efficiency. 
An  enzymatic  process  that  carries  out  the  same 
transformation  as  a chemical  synthesis  pro- 
duces no  side-products  (because  of  an  enzyme's 
high  specificity  to  its  substrate)  or  byproducts 
(because  of  an  enzyme’s  strong  catalytic  power). 
Consequently,  biological  processes  eliminate 
many  conventional  waste  and  disposal  problems 
at  the  front  end  of  the  system— in  the  fer- 
menter. This  high  conversion  efficiency  reduces 
the  costs  of  recycling.  In  addition,  the  efficiency 
of  the  biological  conversion  process  generally 
simplifies  product  recovery,  reducing  capital 
and  operating  costs.  Furthermore,  by  their 
nature,  biologically  based  chemical  processes, 
tend  to  create  some  waste  products  that  are  bio- 
degradable and  valuable  as  sources  of  nutrients. 

Specific  comparisons  of  the  environmental 
hazards  produced  by  conventional  and  biologi- 
cal systems  are  difficult.  Data  detailing  the 
pollution  parameters  for  various  current  chem- 
ical processes  exist,  but  much  less  information 
is  available  for  fermentation  processes,  and  few 


compounds  are  produced  by  both  methods. 
However,  in  most  beverage  distilling  operations, 
pollution  has  been  reduced  to  almost  zero  with 
the  complete  recovery  of  still  slops  as  animal 
feeds  of  high  nutritional  value.  Such  control 
procedures  are  generally  applicable  to  most 
fermentation  processes.  (App.  I-C  describes  the 
pollutants  that  may  he  produced  by  current 
chemical  processes  and  those  expected  from 
biologically  based  processes.) 

The  Environmental  Protection  Agency  has 
estimated  that  the  U.S.  Go\ernment  and  indus- 
try combined  will  spend  o\er  $3(10  billion  to 
control  air  and  watei'  pollution  in  the  decade 
from  1977  through  198P.  Fhe  share'  of  the 
chemical  and  allied  industries  is  about  $2(i  bil- 
lion. Genetic  engineering  technology  may  lu'lp 
alleviate  this  burden  by  offering  cleaner  |)roc- 
esses  of  synthesis  and  better  biological  waste' 
treatment  systems.  The  me)netary  sa\  ings  e'oulel 
be  tremendous.  As  pure  speculation,  if  just  a 
percent  of  the  current  chemical  inelust ry  we're' 
affected,  spending  on  pe)llution  e’e)ulei  l)e  re'- 
duced  by  about  $100  million  per  \ ear. 


Industrial  chemicals  that  may  he  produced 
hy  biological  technologies  


Despite  the  benefits  of  producing  industrial 
chemicals  biologically,  thus  far  major  fermenta- 
tion processes  have  been  developed  primarily 
for  a few  complex  compounds  such  as  enzymes. 
(See  table  11.)  Biological  methods  have  also  been 
developed  for  a few  of  the  simpler  commodity 
chemicals:  ethanol,  butanol,  acetone,  acetic 
acid,  isopropanol,  glycerol,  lactic  acid,  and  citric 
acid. 

Two  questions  are  critical  to  assessing  the 
feasibility  or  desirability  of  producing  various 
chemicals  biologically: 

1.  Which  compounds  can  be  produced  bio- 
logically (at  least  theoretically)? 

2.  Which  compounds  may  be  primarily  de- 
pendent on  genetic  technology,  given  the 
costs  and  availability  of  raw  materials? 


In  principle,  v irtually  all  organic  compounds 
can  be  produced  by  biological  .systt'ins.  If  llu' 
necessary  enzyme  or  enzynu's  arc'  not  know  n to 
exist,  a search  of  the  biological  world  w ill  prob- 
ably uncover  the  appro|)riat('  oiu's.  Alterna- 
tively, at  least  in  theory,  an  ('nzymc'  can  he 
engineered  to  carry  out  tin?  r('(|uirc'd  r('action. 
Within  this  framework,  tiu?  potc'iitial  appc'ars  to 
be  limited  only  by  the?  imagination  ol  the'  l)io- 
technologist— even  though  c('rlain  chc'micals 
that  are  highly  toxic  to  biological  syslc'in.s  are 
probably  not  amenable  to  |)roduclion 

Three  variables  in  particular  afleet  the 
answer  to  the  second  (|U('slion:  iht'  availability 
of  an  organism  or  cMizymc's  for  the'  desired 
transformation;  the  cost  of  tlu'  raw  materi.il: 
and  the  cost  of  the?  production  procc'ss  \\  hen 
specific  organisms  and  production  leehni)lo/;ies 


Ch.5 — The  Chemical  Industry  • 93 


Table  11.— Some  Commercial  Enzymes  and  Their  Uses 

Enzyme 

Source 

Industry  and  application 

Amylase 

Animal  (pancreas) 

Pharmaceutical;  digestive  aids 
Textile:  desizing  agent 

Plant  (barley  malt) 

Baking;  flour  supplement 

Brewing,  distilling,  and  industrial  alcohol:  mashing 
Food:  precooked  baby  foods 
Pharmaceutical:  digestive  aids 
Textile:  desizing  agent 

Fungi  {Aspergillus  niger,  A.  oryzae) 

Baking;  flour  supplement 

Brewing,  distilling,  and  industrial  alcohol:  mashing 
Food:  precooked  baby  foods,  syrup  manufacture 
Pharmaceutical:  digestive  aids 

Bacteria  (Bacillus  subtilis) 

Paper:  starch  coatings 

Starch:  cold-swelling  laundry  starch 

Bromelin 

Plant  (pineapple) 

Food;  meat  tenderizer 
Pharmaceutical:  digestive  aids 

Cellulase  and  hemicellulase  .. 

Fungi  (Aspergillus  niger) 

Food;  preparation  of  liquid  coffee  concentrates 

Dextransucrase 

Bacteria  (Leuconosloc  mesenteroides) 

Pharmaceutical:  preparation  of  blood-plasma 
extenders,  and  dextran  for  other  uses 

Ficin 

Glucose  oxidase  (plus  catalase 

Plant  (fig  latex) 

Pharmaceutical:  debriding  agent 

or  peroxidase) 

Fungi  (Aspergillus  niger) 

Pharmaceutical:  test  paper  for  diabetes 
Food;  glucose  removal  from  egg  solids 

Invertase 

Yeast  (Saccharomyces  cerevisiae) 

Candy:  prevents  granulation  of  sugars  in  soft-center 
candies 

Food:  artificial  honey 

Lactase 

Yeast  (Saccharomyces  fragilis) 

Dairy;  prevents  crystallization  of  lactose  in  ice  cream 
and  concentrated  milk 

Lipase 

Fungi  (Aspergillus  niger) 

Dairy:  flavor  production  in  cheese 

Papain 

Plant  (papaya) 

Brewing:  stabilizes  chill-proof  beer 
Food:  meat  tenderizer 

Pectinase 

Fungi  (Aspergillus  niger) 

Wine  and  fruit  juice:  clarification 

Penicillinase 

Bacteria  (Bacillus  cereus) 

Medicine:  treatment  of  allergic  reaction  to  penicillin, 
diagnostic  agent 

Pepsin 

Animal  (hog  stomach) 

Food:  animal  feed  supplement 

Protease  

Animal  (pancreas) 

Dairy:  prevents  oxidized  flavor 
Food:  protein  hydrolysates 
Leather:  bating 

Pharmaceutical:  digestive  aids 
Textile;  desizing  agent 

Animal  (pepsin) 

Brewing:  beer  stabilizer 

Animal  (rennin,  rennet) 

Dairy:  cheese 

Animal  (trypsin) 

Pharmaceutical:  wound  debridement 

Fungi  (Aspergillus  oryzae) 

Baking:  bread 
Food:  meat  tenderizer 

Bacteria  (Bacillus  subtilis) 

Baking:  modification  of  cracker  dough 
Brewing:  clarifier 

Streptodornase 

Bacteria  (Streptococcus  pyrogenes) 

Pharmaceutical:  wound  debridement 

SOURCE:  David  Perlman.  “The  Fermentation  Industries."  American  Society  lor  Microbiology  News  39:10, 1973,  p.  653. 


have  been  developed,  the  cost  of  raw  materials 
becomes  the  limiting  step  in  production.  If  a 
strain  of  yeast,  for  example,  produces  5 percent 
ethanol  using  sugar  as  a raw  material,  the  proc- 
ess might  become  economically  competitive  if 
tbe  cost  of  sugar  drops  or  the  price  of  petro- 
leum rises.  Even  if  prices  remain  stable,  the 
micro-organisms  might  be  genetically  impro\  ed 
to  increase  their  yield;  genetic  manipulation 
might  soK'e  the  problem  of  an  inefficient 


organism.  Finally,  the  production  process  itself 
is  a factor.  After  fermentation,  the  desired  prod- 
uct must  be  separated  from  the  other  com- 
pounds in  the  reaction  mixture.  As  an  aid  to  re- 
covery, the  production  conditions  might  be 
altered  and  improved  to  generate  more  of  a de- 
sired compound. 

More  than  one  raw  material  can  be  used  in  a 
fermentation  process.  If,  in  the  case  of  ethanol. 


94  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


the  price  of  sucrose  (from  sugarcane  or  sugar 
beets)  is  not  expected  to  change,  the  production 
technology  is  being  run  at  optimum  efficiency, 
and  the  micro-organism  is  producing  as  much 
ethanol  as  it  can,  the  hurdle  to  economic  com- 
petitiveness might  be  overcome  if  a less  expen- 
sive raw  material— cellulose,  perhaps— were 
used.  But  cellulose  cannot  be  used  in  its  natural 
state:  physical,  chemical,  or  biological  methods 
must  be  devised  to  break  it  down  to  its  glucose 
(also  a sugar)  components. 

The  constraints  vary  from  compound  to  com- 
pound. But  even  though  the  role  of  genetics 
must  be  examined  on  a product-by-product  ba- 
sis, certain  generalizations  can  be  made.  Over- 
all, genetic  engineering  will  probably  have  an 
impact  on  three  processes: 

• Aerobic  fermentation,  which  produces  en- 
zymes, vitamins,  pesticides,  growth  regula- 
tors, amino  acids,  nucleic  acids,  and  other 
speciality  chemicals,  is  already  well-estab- 
lished. Its  use  should  continue  to  grow.  Al- 
ready, complex  biochemicals  like  antibiot- 
ics, growth  factors,  and  enzymes  are  made 
by  fermentation.  Amino  acids  and  nucleo- 
tides—somewhat  less  complicated  mole- 
cules—are  sometimes  produced  by  fer- 
mentation. Tbeir  production  is  expected  to 
increase. 

• Anaerobic  fermentation,  which  produces 
organic  acids,  methane,  and  solvents,  is  the 
industry’s  area  of  greatest  current  growth. 
Already,  40  percent  of  the  ethanol  man- 
ufactured in  the  United  States  is  produced 
in  this  way.  The  main  constraint  on  the 
production  of  other  organic  acids  and  sol- 
vents is  the  need  for  cheaper  methods  for 
converting  cellulose  to  fermentable  sugars. 

• Chemical  modification  of  the  fermentation 
products  of  both  aerobic  and  anaerobic 
fermentation,  which  to  date  has  rarely 
been  used  on  a commercial  scale,  is  of 
great  interest.  (See  table  12.)  Cbemical  pro- 
duction technologies  that  employ  high  tem- 
peratures and  pressures  might  be  replaced 
by  biological  technologies  operating  at  at- 
mospheric pressure  and  ambient  tempera- 
ture. A patent  application  has  already  been 
filed  for  the  biological  production  of  one  of 


Table  12. — Expansion  of  Fermentation  Into 
the  Chemical  Industry 


Examples 

Aerobic  fermentation 

Enzymes 

. . Amylases,  proteases 

Vitamins 

. . Riboflavin  Bw 

Pesticides 

. . Bacillus  thuringiensis 

Growth  regulators 

. . Gibberellin 

Amino  acids 

. . Glutamic,  lysine 

Nucleic  acids 

Acids 

. . Malic  acid,  citric  acid 

Anaerobic  fermentation 

Solvents 

. . Ethanol,  acetone,  n-butanol 

Acids 

. . Acetic,  propionic,  acrylic 

SOURCE:  Office  of  Technology  Assessment. 


these  products,  ethylene  glycol,  by  the 
Cetus  Corp.  in  Berkeley,  Calif.  The  procc'ss 
is  claimed  to  he  more  eiK'rgy  etlicif'iit  and 
less  polluting.  If  it  pro\  es  succf’sslul  w Iumi 
run  at  an  industrial  scale,  th(*  tf'chnology 
could  become  significant  to  a U.S.  market 
totaling  $2’/2  billion  |)ci’  v(*ar. 

The  chemical  industry  produce's  a sarif'ty  of 
likely  targets  for  biotechnology.  I'ahles  l-B-27 
through  l-B-32  in  ap|)endi.\  l-B  present  projec- 
tions of  the  potential  economic  impacts  of  ap- 
plied genetics  on  seUntf'd  compounds  that 
rejTresent  large  markets,  and  the  time  frame's 
for  potential  implemientiitiefn.  fable'  l-B-7  lists 
one  large  gremp  e)f  efrganie’  e he'inie’als  tluit  we're' 
identified  by  the  Ceaiefx  Ce)rp.  anel  Massae  hu- 
setts  Institute  e>f 'fe'e:hne)le)gy  (,\iri  ) ;is  ame'iiable' 
to  biotechne)le)gical  pre)elue  lie)n  me'llufels.  flu'y 
are  in  agreement  e)ii  abe)ul  20  pe'iee'iit  e)t  the' 
products  cited,  whie’h  unele'rse'eere's  the*  unee'i- 
tain  nature  e)f  attemipling  le)  pre'elie  t see  lar  inte) 
the  future. 

Fertilizers,  polymers,  mul  pesticides 

Gaseejus  amiiUMiia  is  use'el  te)  pi  efelue  e'  nitrefge'u 
fertilizers.  Ahe)ut  15  hilliefii  tefune's  eil  ammefuia 
were  pre)duceHl  e he'inieeilly  lor  this  purpe»se‘.  in 
1978;  the?  pre)ce?ss  re'e|uire's  kirge'  amefunts  eel 
natural  gas.  Nitre)ge'U  exin  eilse)  he'  e e)in  e'l  teel.  eer 
“fixed,”  te)  amme)iiia  by  e'lizyme's  in  mie  re)-e»i>;a- 
nisms;  ahe)ut  175  billie)n  tefiine's  are*  lixe-el  i)ei 
year,  for  exani|)le',  e)iie'  se|u;ire'  \.irel  e)l  lanel 
planted  w ith  eu'rtain  le'gume's  (sue  h as  se>\  bc.msl 
can  fix  up  te)  2 e)unee's  e)f  nilrefge'ii,  UMUg  hae 


c/7.5 — The  Chemical  Industry  • 95 


tpria  associated  uitli  theii'  roots.  C'liiTently,  mi- 
crobial  production  ot  ammonia  from  nitrogen  is 
not  economically  t'ompetiti\  e.  .Aside  t'rom  the 
dif'ticulties  associated  uitli  the  enzMiie’s  sen- 
sitivity to  owgen  and  the  neai'  total  lack  of 
understanding  of  its  mechanism,  it  takes  the 
e(|ui\  alent  ot  the  energv  in  4 kilograms  (kg)  of 
sugar  to  make  f kg  of  ammonia.  Since  ammonia 
costs  SO.  14  kg  and  sugar  costs  SO. 22  kg,  it  is  un- 
likeK  that  the  chemical  [)rocess  \\  ill  t)e  replaced 
in  the  near  future.  On  the  othei’  hand,  the  genes 
tor  nitrogen  fixation  ha\  e now  t)('en  transferred 
into  veast,  opening  up  tlu*  possil)ilit\  that  agi'i- 
cultui'ally  useful  niti'ogen  can  t)c  made  hv  fer- 
mentation. 

A large  segment  of  tlie  chemical  industry  en- 
gaged in  the  manufacture  of  polymei  s is  shown 
in  table  13.  A total  of  4.3  million  tonnes  of 
fibers,  12  million  tonnes  of  plastics,  and  1.1  mil- 
lion tonnes  of  synthetic  i'ul)t)er  wei'e  produced 
in  the  I'nited  States  in  1078.  All  were  derived 
from  petroleum,  vv  ith  the  e\ce[)tion  of  the  less 
than  1 [lercent  dei'ived  from  cellulose  fibers. 
The  most  likely  ones  are  polyamides  (chemically 
related  to  proteins),  acrylics,  isoprene-type  rub- 
ber, and  polystyrene.  Because  most  monomers, 
the  building  blocks  of  polymers,  are  chemicallv 
simple  and  are  presently  available  in  high  yield 
from  petroleum,  their  microbial  production  in 
the  next  decade  is  unlikelv . 

W hile  hiotechnologv  is  not  ready  to  replace 
the  present  technologv,  its  ev  entual  impact  on 
polvmer  production  will  probably  he  large. 
Biopolvmers  represent  a new  way  of  thinking. 
Most  of  the  important  constituents  of  cells  are 
polymers:  proteins  (polypeptides  from  amino 
acid  monomers),  polvsaccharides  (from  sugar 
monomers),  and  polvnucleotides  (from  nucleo- 
tide monomers).  Since  cells  normally  assemble 
polymers  vv  ith  extreme  specificity,  the  ideal  in- 
dustrial process  would  imitate  the  biological 
production  of  polymers  in  all  possible  respects— 
using  a single  biological  machine  to  convert  a 
raw'  material,  e.g.,  a sugar,  into  the  monomer  to 
polymerize  it,  then  to  form  the  final  product.  A 
more  likely  application  is  the  development  of 
new  monomers  for  specialized  applications. 
Polymer  chemistry  has  largely  consisted  of  the 
study  of  how  their  properties  can  be  modified. 


Table  13.— The  Potential  of  Some  Major  Polymeric 
Materials  for  Production  Using  Biotechnology 


Product 

Domestic  production  1978 
(thousand  tonnes) 

Plastics 

Thermosetting  resins 

Epoxy 

135 

Polyester 

544 

Urea 

504 

Melamine 

90 

Phenolic 

727 

Thermoplastic  resins 

Polyethylene 

Low  density 

3,200 

High  density 

1,890 

Polypropylene 

1 ,380 

Polystyrene 

2,680 

Polyamide,  nylon  type  . . . 

124 

Polyvinyl  alcohol 

57 

Polyvinyl  chloride 

2,575 

Other  vinyl  resins 

88 

Fibers 

Cellulosic  fibers 

Acetate 

139 

Rayon  

269 

Noncellulosic  fibers 

Acrylic 

327 

Nylon 

1,148 

Olefins 

311 

Polyester 

1,710 

Textile  glass 

418 

Other 

7 

Rubbers 

Styrene-butadiene 

628 

Polybutadiene 

170 

Butyl 

69 

Nitrile 

33 

Polychlorophene 

72 

Ethylene-propylene 

78 

Polyisoprene 

62 

SOURCE:  Office  of  Technology  Assessment. 


Conceivably,  biotechnology  could  enable  the 
modification  of  their  function  and  form. 

Pesticides  include  fungicides,  herbicides,  in- 
secticides, rodenticides,  and  related  products 
such  as  plant  growth  regulators,  seed  disinfec- 
tants, soil  conditioners,  and  soil  fumigants.  The 
largest  market  (roughly  $500  million  annually) 
involves  the  chemical  and  microbial  control  of 
insects.  Although  microbial  insecticides  have 
been  around  for  years,  they  comprise  only  5 
percent  of  the  market.  However,  recent  suc- 
cesses in  developing  viruses  and  bacteria  that 
produce  diseases  in  insects,  and  the  negative 
publicity  given  to  chemical  insecticides,  have 
encouraged  the  use  of  microbial  insecticides. 


96  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


or  lli(’  1.’), ()()()  known  species  of  insects,  only 
200  are  harniliil  enongl'i  to  warrant  control  or 
(l('strnction.  fortunately  for  man,  most  of  them 
are  sensitive  to  cei’tain  micro-organisms  which, 
il  they  are  not  toxic  to  man,  nontarget  animals, 
and  |)lants,  can  he  used  as  commercial  insec- 
ticides. 

,\p|)ro\imately  100  known  species  of  bacteria 
are  pathogenic  (disease  causing)  to  insects,  hut 
only  [hvee— Bacillus  popilliae,  B.  thuringiensis 
and  B.  moritai—have  been  developed  into  com- 
mercial insecticides.  6.  popilliae  is  found  and 
produced  only  in  the  larv'ae  of  Japanese  beetles. 


The  other  two  species  can  be  produced  by  con- 
ventional fermentation  techniques.  They  have 
been  useful  because  they  foi'm  spores  that  can 
he  mass-produced  easily  and  are  stable  enough 
to  he  handled  commercially.  The  actual  sub- 
stances that  cause  toxicity  to  the  insect  ai'e  tox- 
ins synthesized  by  the  microbes. 

(ienetic  engineering  should  make  it  possible 
to  construct  more  potent  bacterial  insecticides 
by  increasing  the  dosage  of  the  genes  that  code 
foi'  the  synthesis  of  the  toxins  involved.  Mix- 
tures of  genes  capable  of  directing  the  synthesis 
of  v arious  toxins  might  also  he  pi'oducefl. 


Constraints  on  biological  production  techniques 


The  chief  impediments  to  using  biological 
production  technology  are  associated  with  the 
need  for  biomass.^  They  include: 

• competition  with  food  needs  for  starch  and 
sugar; 

• cyclic  availability; 

• biodegradabilitv  and  associated  storage 
problems; 

• high  moisture  content  for  cellulosics,  and 
high  collection  and  storage  costs; 

• mechanical  processing  for  cellulosics; 

• the  heterogenous  nature  of  cellulosics  (mix- 
tures of  cellulose,  hemicellulose,  and  lig- 
nin); and 

• The  need  for  disposal  of  the  nonferment- 
able  portions  of  the  biomass. 

For  food-related  biomass  sources,  such  as  su- 
gar, corn,  and  sorghum,  few  technological  bar- 
riers exist  for  conv  ersion  to  fermentable  sugars; 
but  subsidies  are  needed  to  make  the  fermenta- 
tion of  sugars  as  profitable  as  their  use  as  food. 
For  cellulosic  biomass  sources  such  as  agricul- 
tural wastes,  municipal  wastes,  and  wood,  tech- 
nological barriers  exist  in  collection,  storage, 
pretreatment,  fermentation,  and  waste  disposal. 
In  addition,  biomass  must  always  be  trans- 
formed into  sugars  by  either  chemical  or  en- 
zymatic processes  before  fermentation  can 
begin. 


■Energy  Emm  Biological  Processes,  op.  oil. 


A second  major  im|)ediment  is  asso('iat(’d 
with  the  purification  stage  of  [iroduction.  Most 
chemical  products  of  fei'inentation  are  pixxsc'nt 
in  extremely  dilute  solutions,  and  concentrating 
these  solutions  to  recovei’  th('  desiixul  product  is 
highly  energy-intensive.  Problems  of  technologv' 
and  cost  will  continue  to  make  this  stage  an  im- 
portant one  to  improve. 

The  developments  in  gi'iK'tics  show  gi’cat 
promise  for  creating  moix*  versatile  micio-orga- 
nisms,  hut  they  do  not  by  themsc'lv cs  pi’oduce  a 
cheaper  fuel  or  plastic.  Associatc'd  technologies 
still  require  more  (?ffici(>nt  f(M'mentation  facil- 
ities and  product  S(q)ai'ation  proc('sses:  mi- 
crobes may  producer  mok'cuU's,  hut  they  will 
not  isolate,  purify,  concc’Uti’ate,  mix,  or  package 
them  foi'  human  us(v 

Fhe  interaction  hetwc'en  genetic  engineering 
and  other  technologi(>s  is  illustrated  by  the 
problems  of  pioducing  ethanol  by  fermenta- 
tion. Fhe  cas(^  study  prc'sc'iiled  in  appendix  ID 
identifi(!s  those  ste|)s  in  the  hiomass-lo-elhanol 
scheiiK?  that  mu'd  t(U'hnological  improvements 
before  the;  |)i'oc(?ss  can  become  ecomunicaf 

Cicnetic  (MigiiKM'iing  is  expected  to  redui c 
costs  in  many  pi'oduction  slj'ps  for  certain 
ones— siK'h  as  the  pretreatmeni  of  the  hiom.iss 
to  make  it  fermentable— gi'iielics  will  |)rnh.ihlv 
not  play  a role:  physical  and  chemic.il  lechnol 
ogies  will  he  responsible  for  tin*  gre.ilest  ad- 
vances. Foi'  otiKM's,  such  as  distillation  ^jenelic 


Ch.  5— The  Chemical  Industry  • 97 


U’c'linolo^it's  should  make  il  |)ossihle  lo  engineer 
organisms  that  ran  ferment  at  liigli  tempera- 
tures (82°  to  85°  (')  so  that  tlie  fermentation  and 
at  least  part  of  the  distillation  can  both  take 
place  in  the  same  reactor.*  Various  technol- 

' I hi-rmophilic  t'lhiiiuil  |ii'U(liK  t‘r.'<  h.iu'  ,ilriM(l\  hi'cn  (Icm  i iIxhI 
ill  ihr  ucmiN  t lostridiiim  li.iv  tht-rmoi  rlliim)  In  acldilinn.  Hi'niMi- 
r.tIK  (•nfiini‘rti'<l  ilt'sn'ihnil  a>  a cms.^  hclwcfn  mmsI.s 

.uul  lIuM'iiuiphilic  l>a('U'na  can  li'rnicnl  at  70°  C 


An  overview  of  impacts 

I'he  cost  of  raw  materials  may  become  cheap- 
er than  the  petroleum  now  used— especially  if 
cellulose  con\  ersion  technologies  can  he  de\el- 
oped,  I'he  source  of  raw  materials  would  also 
he  broader  since  se\  eral  kinds  of  biomass  could 
he  interchanged,  if  necessaiy.  I’oi'  small  (|uan- 
tities  of  chemicals,  the  I'aw  material  su|)ply 
would  he  more  dependable,  particularly  be- 
cause of  the  domestic  supph  of  available  bio- 
mass. For  substances  produced  in  large  quanti- 
ties, such  as  ethanol,  the  su[)ply  of  biomass 
could  limit  the  usefulness  of  hiotechnologv. 

Raw  materials,  such  as  organic  wastes,  could 
he  piocessed  both  to  produce  products  and 
reduce  pollution.  .Nevertheless,  the  impact  on 
total  imported  petroleum  w ill  he  low . Estimates 
of  the  current  consumption  of  petroleum  as  a 
raw  material  for  industrial  chemicals  is  appro.x- 
imately  5 to  8 percent  of  the  total  imported. 

Impacts  on  the  process  include  relatively 
cheaper  production  costs  for  selected  com- 
pounds. For  these,  lower  temperatures  and 
pressures  can  be  used,  suggesting  that  the  proc- 
esses might  be  safer.  Chemical  pollution  from 
hiotechnologv'  may  be  lower,  although  methods 
of  disposal  or  new  uses  must  be  found  for  the 
micro-organisms  used  in  fermentation.  Finally, 
the  biological  processes  will  demand  the  devel- 
opment of  new  technologies  for  the  separation 
and  purification  of  the  products. 

Impacts  on  the  products  include  both  cheaper 
existing  chemicals  as  well  as  entirely  new  prod- 
ucts. Since  biotechnology  is  the  method  of 
choice  for  producing  enzymes,  new  uses  for  en- 


ogies,  such  as  the  immobilization  of  whole  cells 
in  reactoi'  columns,  could  he  tleveloped  in  paral- 
lel vv  ith  genetic  technologies  to  increase  the  sta- 
hilitv  of  cells  in  fermenters. 


rlu‘  iiiK  ol  such  ihcmiophilic  leniu'iilations  are  sif'iiili- 

cani:  lernienlalion  lime  is  I'unsiderahlv  I'eiluced:  Ihe  risk  ol  con- 
lamination  is  nearly  eliminated:  and  cooling  re(|uirements  are 
lower  due  lo  Ihe  high(-r  temperature  ol'  the  rermenting  hioth. 


zv  Hies  may  expand  and  drive  this  sector  of  the 
industry. 

Impacts  on  other  industries 

.Although  genetic  engineering  will  develop 
new  techniques  for  synthesizing  many  sub- 
stances, the  direct  displacement  of  any  present 
industry  appears  to  he  doubtful:  Genetic  engi- 
neering should  he  considered  simply  another  in- 
dusti'ial  tool.  As  such,  any  industry's  response 
should  he  to  use  this  technique  to  maintain  its 
positions  in  its  respectiv  e markets.  The  point  is 
illustrated  by  the  variety  of  companies  in  the 
pharmaceutical,  chemical,  and  energy  indus- 
tries that  have  invested  in  or  contracted  with 
genetic  engineering  firms.  Some  large  com- 
panies are  already  developing  inhouse  genetic 
engineering  research  capabilities. 

The  frequent,  popular  reference  to  the  small, 
innovative  “genetic  engineering  companies”  as  a 
major  new  industry  is  somewhat  misleading. 
The  companies  (see  table  14)  arose  primarily  to 
convert  micro-organisms  with  little  commercial 
use  into  micro-organisms  with  commercial  po- 
tential. A company  such  as  the  Cetus  Corp.  ini- 
tially used  mutation  and  selection  to  improve 
strains,  whereas  other  pioneers  such  as  Genen- 
tech,  Inc.,  Biogen,  S.  A.,  and  Genex  Corp.  were 
founded  to  exploit  recombinant  DNA  (rDNA) 
technology.  Part  of  their  marketing  strategy  in- 
cludes the  sale  or  licensing  of  genetically  engi- 
neered organisms  to  large  established  commer- 
cial producers  in  the  chemical,  pharmaceutical, 
food,  energy,  and  mining  industries.  Each  engi- 


98  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Table  14.- 

—Some  Private  Companies  With  Biotechnology  Programs 

Company 

Founded 

Approximate 
employees  1979 

Ph.  D.s  1979 

Research  capacity 
Recombinant  DNA  Hybridomas 

Atlantic  Antibodies 

1973 

50 

2 

X 

Bethesda  Research  Laboratories  . . 

1976 

130 

30 

X 

X 

Biogen 

1978 

30  (50q 

(-|8b)(3)(5) 

X 

Xb 

Bens  Bio  Logicals 

1979 

15 

10 

X 

X 

Centocor 

1979 

200-  low 

? 

X 

Cetus  

1972 

250 

50 

X 

X 

Clonal  Research 

1979 

6 

1 

X 

Collaborative  Researcht^ 

1961 

85 

15 

X 

X 

(Collaborative  Genetics) 

(1979) 

(4) 

(3) 

X 

Genentech  

1976 

90 

30 

X 

Genex  

1977 

30 

12 

X 

Hybritech 

1978 

33(1) 

6 

X 

Molecular  Genetics 

1979 

6(4) 

2 

X 

Xb 

Monoclonal  Antibodies 

1979 

6 

3 

X 

New  England  Biolabs 

1974 

22(22) . 5(4) 

? 

X 

®F.  Eberstadt  & Co.  estimates. 

^Expected  by  December  1980. 

''Collaborative  Research  is  a major  owner  of  Collaborative  Genetics.  The  division  between  them  is  not  yet  distinct. 

SOURCES:  (1)  Science  208,  p.  692-693, 1980  (52  people  to  expand  to  100  by  1981). 

(2)  Science  208,  p.  692-693, 1980  (20  senior  persons). 

(3)  Science  208,  p.  692-693, 1980  (16  scientists,  30  employees). 

(4)  Dun  & Bradstreet,  Inc. 

(5)  Chemical  and  Engineering  News,  Mar.  19, 1980. 

Office  of  Technology  Assessment. 

neering  firm  also  intends  to  manufacture  some 
products  itself.  It  is  likely  that  the  products  re- 
served for  inhouse  manufacture  will  he  low- 
volume,  high-priced  compounds  like  interferon. 

Genetic  engineering  by  itself  is  a relatively 
small-scale  laboratory  operation.  Consequently, 
genetic  engineering  firms  will  continue  to  offer 
services  to  companies  that  do  not  intend  to 
develop  this  capacity  in  their  own  inhouse  lab- 
oratories. Specifically,  a genetic  engineering 
company  may  contract  with  a firm  to  develop  a 
biological  production  method  for  its  products. 

At  the  same  time,  larger  companies  might  estab- 
lish inhouse  staffs  to  develop  biological  methods 
for  both  old  and  new  products.  (Several  larger 
companies  already  have  more  inhouse  genetic 
engineering  personnel  than  some  of  the  inde- 
pendent genetic  engineering  companies.) 

In  addition,  suppliers  of  genetic  raw  materials 
may  decide  to  expand  into  the  production  of 
genetically  engineered  organisms.  Suppliers  of 
restriction  endonuclease  enzymes  for  example, 
which  are  used  in  constructing  rDNA,  have 
already  entered  the  field.  Diagnostic  firms  could 
develop  new  bioassays  for  which  they  them- 
selves would  guarantee  a market.  Finally,  com- 
panies with  byproducts  or  waste  products  are 


beginning  to  examine  the  possibility  of  c'om  (M  l- 
ing  them  into  useful  products.  This  approacli 
(which  is  somewhat  moi  e developc’d  in  liurope) 
assumes  that  with  the  propc'r  technology  the 
waste  materials  can  become  a resource. 

Some  industries,  including  manulacturcM's  of 
agitators  (drives),  centrifuges,  e\  a|)orators,  ler- 
menters,  dryers,  storage  tanks  and  process 
vessels,  and  conti'ol  and  instruim'ntation  sys- 
tems, might  profit  by  |)roducing  e(|uipm('ul 
associated  with  fei'inentation. 

Impacts  on  university  resiutrt'h 

From  the  beginning,  genetic  (Migineeiing 
firms  established  strong  ties  with  uni\ crsities 
These  were  responsible  foi'  pio\  iding  most  of 
the  scientific  knowledges  that  formed  the  basis 
for  applied  genetics  as  well  as  the  initial  scien- 
tific workforce: 

• CetLis  Cioi'p.  (sstahlislu'd  a pattern  by  ic- 
cruiting  a prestigious  hoard  of  Scientific 
Advisors  who  re'inain  in  academic  posi 
tions. 

• Genentech,  Inc.,  cofounde'd  In  a profe-ssor 
at  the  University  of  Galifornia  at  San  t r.in- 


Ch.  5— The  Chemical  Industry  • 99 


cisco,  initially  (Ifpendecl  largely  on  outside 
scientists. 

• Biogen,  S.  \..  was  organized  In  prolessors 
at  Har\ard  and  MM  plus  six  Kuropean  sci- 
entists. and  placed  RiSt  1)  contracts  w ith  aca- 
ch'iiiic  researchers. 

• ('ollahoratix  e (i('netics  has  a .\ohel  prize 
w inner  from  Mi  l as  the  chairman  ot  its  sci- 
entitic  ad\  isor\  hoard. 

• Ih  hritech.  Inc.,  has  as  its  scientific  nucleus 
a UniwrsitN  of  ('alilornia,  .San  Diego,  pro- 
fessor complemented  In  scientists  at  the 
Salk  Institute. 

In  addition  to  the.se  companies,  otluM's  ha\  e also 
been  establishing  clo.ser  ties  with  the  academic 
community. 

Much  of  the  research  that  will  he  u.seful  to  in- 
dustry  w ill  continue  to  he  carried  out  in  uni\  er- 
sity  laboratories.  .\t  ()res(Mit,  it  is  often  difficult 
to  decide  w helher  a re.search  |)roject  should  he 
classifieil  as  "l)a.sic"  (generally  more  interesting 
to  an  academic  I'e.searcherl  oi'  "applied  " (gener- 
ally more  interesting  to  industry).  E.g.,  a change 
in  the  genetic  code,  w hich  increases  gene  acti\  i- 
ty,  would  be  just  as  exciting  to  a basic  scientist 
as  to  an  industrial  one. 

This  dialog  between  tbe  uni\ersities  and  in- 
dustry—both  through  formal  and  informal  ar- 
rangements—has  fostered  inno\ation.  .Although 
the  number  of  patents  applied  for  is  not  a direct 
reflection  of  tbe  le\  el  of  inno\  ation,  it  is  still  one 
indication.  B\-  the  end  of  1980,  several  hundred 
patent  applications  were  filed  for  genetically 
engineered  micro-organisms,  their  products, 
and  their  processes. 

I'niversity  research  has  clearly  affected  in- 
dustrial development,  and  has  in  turn  been  af- 
fected by  industry.  .Although  the  benefits  are 
easily  recognized,  some  drawbacks  have  been 
suggested.  The  most  serious  is  the  concern  that 
univ  ersity  scientists  will  be  restrained  in  their 
academic  pursuits  and  in  their  exchange  of  in- 
formation and  research  material.  To  date,  anec- 
dotal information  suggests  that  some  scientists 
are  being  more  circumspect  about  sharing  in- 
formation. Still,  secrecy  is  not  new  to  highly 
competitive  areas  of  biomedical  research.  In  ad- 
dition, scientists  in  other  academic  disciplines 


u.seful  (o  incluslrv— such  as  clu'misiry  and  phys- 
ics-have manag('d  lo  achicnc  a halaiK'c  he- 
(ween  secrecv  and  openiK'ss. 

77if  sfH'iul  impiu'ts  nf  local 
iiulustrial  acti city 

D('spii(‘  the  extensive  media  (U)verage  of 
rl)\.A  and  other  forms  ol  geiuUic  engim?ering, 
there  is  little  ev  ick'ucc'  that  peopU?  vv  ho  liv  e near 
companii's  using  such  t('chni(|ues  are  still  great- 
ly concerned  about  possihU*  hazards.  This  may 
he  partly  owing  to  a lack  of  awareness  that  a 
particular  companv  is  doing  g(>n(>tic  r(?search 
and  partly  he('aus(‘  companies  thus  far  have 
adhered  to  the  National  lnstitut(?s  of  Health 
(N'llll  Guidelines.  Some  compani(‘s  hav(?  |)laced 
individuals  on  theii’  institutional  biosafety  com- 
mittees who  ai'(>  res|)(*cted  and  trusted  mem- 
bers of  the  local  community.  Ry  involving  the 
local  citizens  with  no  vested  cor|)orate  interest, 
a mechanism  for  oversight  has  he(Mi  provided. 
(For  a moi'e  detailed  discussion,  see  ch.  1 1.) 

Impacts  on  manpower 

I'wo  tv  [)es  of  impacts  on  vvorkei’s  can  he  ex- 
[)ected: 

• The  creation  of  jobs  that  replace  those  held 
by  others.  E.g.,  a worker  involved  in 
chemical  production  might  be  replaced  by 
one  producing  the  same  product  biologi- 
cally. 

• The  creation  of  new  jobs. 

Workers  in  three  categories  would  be  af- 
fected: 

• those  actually  involv  ed  in  the  fermentation- 
production  phase  of  the  industry; 

• those  inv  olved  in  the  R&.D  phase  of  the  in- 
dustry, particularly  professionals;  and 

• those  in  support  industries. 

Projections  of  manpower  requirements  are 
only  as  accurate  as  the  projections  of  the  level  of 
industrial  activity.  In  the  past  5 years,  about  750 
new  jobs  hav  e been  created  within  the  small  ge- 
netic engineering  firms  (including  monoclonal 
antibody  producers).  Of  these,  approximately 
one-third  hold  Ph.  D.  degrees. 


100  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


Data  obtained  through  an  OTA  survey  of  284 
firms  indicate  that  the  pharmaceutical  industry 
employs  the  major  share  of  personnel  working 
in  applied  genetics  programs.  (See  table  15.)  The 
average  number  of  Ph.  D.s  in  each  industry  is 
given  in  table  16.  A rough  estimate  of  profes- 
sional scientific  manpower  at  this  level  includes: 
6 in  food,  45  in  chemical,  120  in  pharmaceutical, 
and  18  in  specialty  chemicals— a total  of  189.  If 
the  number  of  research  support  personnel  is 
approximately  twice  the  number  of  Ph.  D.s,  the 
total  rises  to  about  570.  If  $165,000  per  year  is 
required  to  support  one  Ph.  D.  in  industry,  the 
total  value  of  such  manpower  is  approximately 
$31  million. 

Estimates  of  the  number  of  companies  en- 
gaged in  applied  genetics  work  in  1980  can  be 
compared  with  the  total  number  of  firms  with 
fermentation  activities.  A tabulation  of  firms  on 
a worldwide  basis  in  1977  revealed  145  com- 
panies, of  which  27  were  American.  (See  table 
17.)  These  companies  produced  antibiotics,  en- 
zymes, solvents,  vitamins  and  growth  factors. 


Table  15.— Distribution  of  Applied  Genetics 
Activity  in  Industry 


Classification 

Distribution  of  applied 
genetics  activity  by 
company  classa 

Percent 
of  total 

Food 

(6/46) 

13 

Chemical 

(9/52) 

17 

Pharmaceutical .... 

(12/25) 

48 

Specialty  chemical^ 

(6/68) 

9 

^Ignores  small  firms  specializing  in  genetic  research. 
'’Food  ingredients,  reagents,  enzymes. 

SOURCE:  Office  of  Technology  Assessment. 

Table  16.— Manpower  (low-(average)-high)  Distribution 
of  a Firm  With  Applied  Genetics  Activity 


Ph.  D.  M.S.  Bachelors 

Food 0-(1)-2  0-(1)-2  0-(2)-8 

Chemical 3-(5)-7  0-(1)-2  2-(5)-7 

Pharmaceutical 2-(10)-24  1-(4)-9  1-(8)-20 

Specialty 1-(3)-8  1-(3)-4  2-(2)-4 

Biotechnology 

Genetic  engineering.  3-(15)-32  2-(11)-20  5-(15)-25 

Hybridoma 1-(3)-6  0-(2)-0  0-(20)-0 

Other 0-(2)-4  2-(4)-6  8-{10)-13 

Average 1-(6)-12  1-(4)-6  3-(8)-12 


Table  17.— Index  to  Fermentation  Companies 

1.  Abbott  Laboratories,  North  Chicago,  III. 

2.  American  Cyanamid,  Wayne,  N.J. 

3.  Anheuser-Busch,  Inc.,  St.  Louis,  Mo. 

4.  Bristol-Myers  Co.,  Syracuse,  N.Y. 

5.  Clinton  Corn  Processing  Co.,  Clinton,  Iowa 

6.  CPC  International,  Inc.,  Argo,  III. 

7.  Dairyland  Laboratories,  Inc.,  Waukesha,  Wis. 

8.  Dawe’s  Laboratories,  Inc.,  Chicago  Heights,  III. 

9.  Grain  Processing  Corp.,  Muscatine,  Iowa 

10.  Hoffman-LaRoche,  Inc.,  Nutley,  N.J. 

11.  IMC  Chemical  Group,  Inc.,  Terre  Haute,  Ind. 

12.  Eli  Lilly  & Co.,  Indianapolis,  Ind. 

13.  Merck  & Co.,  Inc.,  Rahway,  N.J. 

14.  Miles  Laboratories,  Inc.,  Elkhart.  Ind. 

15.  Parke,  Davis  & Co.,  Detroit,  Mich. 

16.  S.  B.  Penick  & Co.,  Lyndhurst,  N.J. 

17.  Pfizer,  Inc.,  New  York,  N.Y. 

18.  Premier  Malt  Products,  Inc.,  Milwaukee,  Wis. 

19.  Rachelle  Laboratories,  Inc,,  Long  Beach,  Calif. 

20.  Rohm  & Haas,  Philadelphia,  Pa. 

21.  Sobering  Corp.,  Bloomfield,  N.J. 

22.  G.  D.  Searle  & Co.,  Skokie,  III. 

23.  E.  R.  Squibb  & Sons,  Inc,,  Princeton,  N.J. 

24.  Standard  Brands,  Inc.,  New  York,  N.Y. 

25.  Stauffer  Chemical  Co.,  Westport.  Conn. 

26.  Universal  Foods  Corp.,  Milwaukee,  Wis. 

27.  The  Upjohn  Co.,  Kalamazoo,  Mich. 

28.  Wallerstein  Laboratories,  Inc.,  Morton  Grove.  III. 

29.  Wyeth  Laboratories,  Philadelphia.  Pa. 

SOURCE:  Office  of  Technology  Assessment. 

nucleo.side.s,  amino  acid.s,  and  mi.sccllancou.s 
product.s.  (Son  tahU*  18.)  I ho  only  rhnniral  lirm 
listed  was  th(>  Stauffer  ( iK'mieal  ( o.  Ten  lirms 
are  listed  as  ha\  ing  the  ahilitx  to  product*  food 
and  feed  yeast.  (See  table  If).)  ( orrecling  lor 
firms  listed  Iwictf,  at  It'ast  .38  I'.S  firms  were 
engaged  in  significant  fermentation  acti\il\  lor 
commercial  products,  ('xcluding  alcoholic  he\ 
erages,  in  1977.  i\ot  all  ha\  (*  research  expertise 
in  fermentation  or  biotechnology,  much  less  a 
regular  genetics  program:  10  to  2(1  were  in  the 
chemical  industry:  25  to  40  in  lermenlalion  (en- 
zyme, |)harmac(*utical,  lood,  and  specialized 
chemicals);  and  10  to  15  in  hiotechnologx  (genet 
ic  engintM'i’ing)— or  about  45  to  75  I inns  in  all 

If  ax  eragtf  manpower  numbers  .ire  used  the 
total  numh(*r  ol  |)rolessionals  iiuohed  in  com 
mereial  applit'd  genetics  rt'search  is: 

I’ll.  I)..s:  30(1-4.10 
Others: 

900- 1, 3.10 

rhe  number  ol  workers  that  will  lie  on  ol\  cd 
in  the  production  phast*  ol  hiotechnologx  rcpie 


SOURCE:  Office  of  Technology  Assessment. 


Ch.5 — The  Chemical  Industry  • 101 


Table  18.— Fermentation  Products  and  Producers 


Product  Some  producers*  Product  Some  producers* 


Amino  acids 

Lalanine 

L-arginine 

L-aspartic  acid  . . . 

L-citrulline 

Lglutamic  acid 25 

L-glutamine 

L-glutathione 

L-histidine 

L-homoserine 

L-isoleucine 

L-leucine 

L-lysine 

L-methionine 

L-ornithine 

L-phenylalanine 

Lproline 

Lserine 

L-threonine 

L-tryptophan 

L-tyrosine 

L-valine . 

Miscellaneous  products  and  processes 


Acetoin  

Acyloin 13 

Anka-pigment  (red) 

Blue  cheese  flavor 7 

Desferrioxamine 

Dihydroxyacetone 17,21,28 

Dextran 


Diacetyl  (from  acetoin) 

Ergocornine 

Ergocristine 

Ergocryptine 

Ergometrine 

Ergotamine 

Bacillus  thuringiensis  insecticide 1 


Lysergic  acid 

Paspalicacid 

Picibanil 

Ribose 

Scleroglucan 

Sorbose  (from  sorbitol) 10,17 

Starter  cultures 7,13,14 

Sterol  oxidations 22,27 

Steroid  oxidations 21,23,27,29 

Xanthan 13,17 

Antibiotics 

Adriamycin 

Amphomycin 

Amphotericin  B 23 

Avoparcin 2 

Azalomycin  F 

Bacitracin 11,16,17 


Bambermycins 

Bicyclomycin 

Blasticidin  S 

Bleomycin 

Cactinomycin 

Candicidin  B 16 

Candidin 


Capreomycin 

Cephalosporins 4,12 

Chromomycin  A> 

Colistin 

Cycloheximide 27 

Cycloserine 11 

Dactinomycin 13 

Daunorubicin 

Destomycin 

Enduracidin 

Erythromycin 17,27 


Fortimicins. 
Fumagillin  . 
Fungimycin 
Fusidic  acid 


Gentamicins 21 

Gramicidin  A 28 

Gramicidin  J (S) 

Griseofulvin 

Hygromycin  B 12 

Josamycin 

Kanamycins 4 

Kasugamycin 

Kitasatamycin 

Lasalocid 10 

Lincomycin 27 


Lividomycin  . . 
Macarbomycin 
Mepartricin. . . 
Midecamycin  . 
Mikamycins  . . 


Mithramycin 17 

Mitomycin  C 4 

Mocimycin 

Monensin 

Myxin 10 

Neomycins 16,17,23,27 

Novobiocin 27 

Nystatin 23 

Oleandomycin 17 

Oligomycin 

Paromomycins 15 

Penicillin  G 4,12,13,17,23,29 

Penicillin  V 1,4,12,17,23,29 

Penicillins  (semisynthetic) 4,13,17,23,29 

Pentamycin 

Pimaricin 

Polymyxins 17 


Polyoxins  .... 
Pristinamycins 
Quebemycin. . 
Ribostamycin. 
Rifamycins . . . 
Sagamicin.  . . . 
Salinomycin . . 

Siccanin 

Siomycin 


Sisomicin 21 

Spectinomycin 27 

Streptomycins 13,17,29 

Tetracyclines 

Clortetracycline 19 


102  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Table  18.— Fermentation  Products  and  Producers 


Product Some  producers^ Product Some  producers^ 


Demeclocycline 2 

Oxytetracycline 17,19 

Tetracycline 2,4,17,19,23,27 

Tetranectin 

Thiopeptin 

Thiostrepton 23 

Tobramycin 12 

Trichomycin 

Tylosin 12 

Tyrothricin 16,28 

Tyrocidine 

Uromycin 

Validamycin 

Vancomycin 12 

Variotin 

Viomycin 

Virginiamycin 

Enzymes 


Amylases 5,19,20,24,28 

Amyloglucosidase 5,6,14,28 

Anticyanase 

L-asparaginase 

Catalase 8,14 

Cellulase 6,20,28 

Dextranase 

‘Diagnostic  enzymes’ 

Esterase-lipase 28 

Glucanase 28 

Glucose  dehydrogenase 

Glucose  isomerase 3,5,14,24 

Glucose  oxidase 8,14 

Glutamic  decarboxylase 18 

Hemi-cellulase 14,20,28 

Hespiriginase 

Invertase 24,26,28 


Lactase 28 

Lipase 20 

Microbial  rennet 17,28 

Naringinase 28 

Pectinase 20,28 

Pentosanase 20,28 

Proteases 14,17,18,20,28 

Streptokinase-streptodornase 2 

Uricase 

Organic  acids 

Citric  acid 14,17 

Comenicacid 17 

Erythorbicacid 

Giuconicacid 4,17,18 

Itaconicacid 17 

2-keto-D-giuconic  acid 17 

a-ketoglutaric  acid 

Lactic  acid 5 

Malic  acid 

Urocanic  acid 

Solvents 

Ethanol 9 

2,3-butanediol 

Vitamins  and  growth  factors 

Gibberellins 1,12,13 

Riboflavin 13 

Vitamin  Bi2 13 

Zearalanol 11 

Nucleosides  and  nucleotides 

5- ribonucleotides  and  nucleosides 

Orotic  acid 

Ara-A-(9-/3-D-arabino-furanosyl) 15 

6- azauridine 


®Blank  means  no  U.S.  producer  in  1977;  therefore,  is  produced  by  one  or  more  foreign  firms  (from  at  least  120  different  firms). 
SOURCE:  Office  of  Technology  Assessment. 


sents  a major  impact  of  genetic  engineering.  To 
estimate  this  number  these  two  calculations 
must  be  made; 

• the  value  or  volume  of  chemicals  that 
might  be  produced  by  fermentation,  and 

• the  number  of  production  workers  needed 
per  unit  volume  of  chemicals  produced. 

Any  prediction  of  the  potential  volume  of 
chemicals  is  necessarily  filled  with  uncertain- 
ties. The  approximate  market  value  of  organic 
chemicals  produced  in  the  United  States  is  given 
in  appendix  I-B.  Total  U.S.  sales  in  1979  were 
calculated  to  be  over  $42  billion.  On  the  basis  of 
the  assumptions  made,  $522  million  worth  of 
bulk  organic  chemicals  could  be  commercially 


produced  by  genetically  engiiK'cred  strains  in 
10  years  and  $7.1  billion  in  20  years,  fable 
I-B-10  in  appendix  I-B  lists  the  potential  markets 
for  pharmaceuticals.  I'.xcluding  nuMhane  |)ro- 
duction,  the  total  |)otential  market  lor  products 
obtained  from  genetically  engineered  orga- 
nisms is  approximately  $ l-t.O  billion. 

If  the  production  of  chemicals  having  this 
value  is  carried  out  by  h'rmenlalion.  it  is  possi- 
ble to  calculate  how  many  workers  will  he 
needed.  Data  obtained  from  industrial  sources 
reveal  that  2 to  5 workers,  including  those  in 
supervision,  services,  and  production  .ire  re- 
(|uired  foi'  $1  million  worth  of  product  Hem  e 
30,000  to  75,000  workers  would  he  |■e(|uired  loi‘ 
the  estimated  $ 14.0  hillion  market 


Ch.  5— The  Chemical  Industry  • 103 


Table  19.— U.S.  Fermentation  Companies 


Producers  ol  Baker’s  yeast  and  food/feed  yeast  in 
the  United  States  in  1977 
Baker’s  yeast: 

American  Yeast  Co.,  Baltimore.  Md. 
Anheuser-Busch,  Inc.,  St.  Louis,  Mo. 

Federal  Yeast  Co.  (now  Diamond  Shamrock), 
Baltimore.  Md. 

Fleischmann  Yeast  Co.,  New  York,  N Y. 
Universal  Foods  Corp..  Milwaukee.  Wis. 
Food/feed  yeast: 

Amber  Laboratories,  Juneau,  Wis. 

Amoco  Foods  Co..  Chicago,  III. 

Boise-Cascade,  Inc.,  Portland.  Oreg. 

Diamond  Mills.  Inc..  Cedar  Rapids,  Iowa 
Fleischmann  Yeast  Co.,  New  York.  N Y, 

Lakes  States  Yeast  Co.,  Rhinelander,  Wis. 
Stauffer  Chemical  Co..  Westport.  Conn, 

Enzyme  producers,  1977 

Clinton  Corn  Processing  Co.,  Clinton,  Iowa 

Miles  Laboratories,  Inc.,  Elkhart.  Ind. 

Premier  Malt  Products,  Inc.,  Milwaukee,  Wis. 


SOURCE:  Compiled  by  Perlman.  American  Society  lor  Microbiology  News  43:2. 

1977,  pp  82-89 

Since  the  chemicals  considered  above  are 
ciirrenth-  l)eiiif4  |)roduced,  any  new  jobs  in  bio- 
tecbnolog^^'  will  displace  the  old  ones  in  the 
chemical  itidiistr\  . V\  betber  the  change  will  re- 
sult in  a net  loss  or  gain  in  the  number  of  jobs  is 
difficult  to  predict.  Howe\er,  a rough  estimate 


indicates  that  appro.ximately  the  same  number 
of  workei's  will  be  retjuired  per  unit  of  output. 

Kstimates  of  the  number  of  workers  are  di- 
\ ided  into:  1)  workers  directly  iiwoh  ed  in  the 
giowth  of  the  organisms;  and  2)  workers  in- 
\()l\ed  in  the  '‘reco\ery”  phase,  where  the 
organisms  are  bar\  ested  and  the  chemical  prod- 
uct is  e.xtracted,  pui'ified,  and  packaged.  Based 
on  industry  data,  the  number  of  workei's  in  the 
fermeiitation  phase  is  approximately  30  percent 
of  the  total,  and  those  in  reco\  erv  approximate- 
ly 50  percent.  Hence,  about  9,000  to  22,500 
workers  might  he  expected  to  hold  jobs  in  the 
immediate  fermentation  area,  and  about  15,000 
to  37,500  workers  would  he  in\'ol\ed  in  han- 
dling the  production  mediLim  (with  or  without 
the  oi'ganisms). 

Estimates  of  the  number  of  totally  new'  jobs 
that  would  be  created  are  highly  speculative; 
they  should  allow  for  estimates  of  increases  in 
the  quantity  of  chemicals  currently  being  pro- 
duced and  the  production  of  totally  new  com- 
pounds. According  to  estimates  by  Genex,  the 
new  and  growth  markets  may  reach  $26  billion 
by  the  year  2000,  which  would  add  52,000  to 
130,000  jobs  to  tbe  present  number. 


chapter  6 

The  Food  Processing  Industry 


Chapter  6 


Page 

Introduction— The  Industry 107 

Single-Cell  Protein 107 

Genetic  Engineering  and  SCP  Production 109 

Commercial  Production 109 

Genetics  in  Backing,  Brewing,  and  Winemaking  . 110 

Microbial  Polysaccharides Ill 

Enzymes Ill 

Genetic  Engineering  and  Enzymes  in  the 

Food  Processing  Industry 112 

Sweeteners,  Flavors,  and  Fragrances 112 

Overview 113 


Tables 


21.  Classification  of  Yeast-Related  U.S.  Patents  . . 108 

22.  Comparison  of  Selling  Price  Ranges  for 
Selected  Microbial,  Plant,  and  Animal 

Protein  Products 108 

23.  Raw  Materials  Already  Tested  on  a 

Laboratory  or  Small  Plant  Scale 109 

Figure 

Figure  No.  Page 

26.  The  Use  of  Hybridization  To  Obtain  a Yeast 
Strain  for  the  Production  of  Low- 
Carbohydrate  Beer  110 


Table  No.  Page 

20.  Estimated  Annual  Yeast  Production,  1977  . . . 108 


Chapter  6 

The  Food  Processing  Industry 


Introduction — the  industry 


The  food  processing  inclustrv  comprises 
those  manutacturers  that  transform  or  process 
agricultural  products  into  edil)le  products  foi' 
market.  It  is  distinguished  from  the  pioduction, 
or  farming  and  breeding  [)ortions  of  the  agricul- 
tural industry. 

(lenetics  can  he  used  in  the  food  processing 
industry  in  two  ways;  to  design  micro-orga- 
nisms that  transform  inedible  biomass  into  food 
for  human  consumf)tion  or  for  animal  feed:  and 
to  design  organisms  that  aid  in  food  processing, 
either  by  acting  dii'ecth’  on  the  food  itself  or  by 
prox  iding  materials  that  can  he  added  to  food. 

Eight  million  to  ten  million  people  work  in  the 
meat,  poultry,  dairx'.  and  baking  industries:  in 
canned,  cured,  and  frozen  food  plants;  and  in 
mov  ing  food  from  the  farm  to  the  dinner  table. 
In  1979,  the  payroll  was  ox  er  S3. 2 billion  for  the 
meat  and  poultrx  industries,  S2.6  billion  for 
baking,  and  $1.9  billion  for  food  processing. 


Single-cell  protein  

The  interest  in  augmenting  the  xxorld's  sup- 
ply of  protein  has  focused  attention  on  micro- 
bial sources  of  protein  as  food  for  both  animals 
and  humans.*  Since  a large  portion  of  each 
bacterial  or  yeast  cell  consists  of  proteins  (up  to 
72  percent  for  some  protein-rich  cells),  large 
numbers  hax  e been  groxx  n to  supply  single-cell 
protein  (SCP)  for  consumption.  The  protein  can 
be  consumed  directly  as  part  of  the  cell  itself  or 
can  be  extracted  and  processed  into  fibers  or 
meat-like  items.  By  noxx',  adx  anced  food  proc- 
essing technologies  can  combine  this  protein 
xvith  meat  flaxoring  and  other  substances  to 
produce  nutritious  food  that  looks,  feels,  and 
tastes  like  meat. 

*.As  an  e.xample  of  the  potential  significance  of  SCP.  the  So\  iet 
Union,  which  is  one  of  the  largest  producers,  e.xpects  to  produce 
enough  fodder  yeast  from  internally  at  ailable  raw  materials  to  be 
self-sufficient  in  animal  protein  foodstuffs  by  1990. 


Traditionally,  micro-organisms  haxe  been 
usetl  to  stabilize,  flaxor,  and  modify  various 
properties  of  food.  More  recently,  efforts  have 
been  made  to  control  microbial  spoilage  and  to 
ensure  that  foods  are  free  from  micro-orga- 
nisms that  may  he  hazardous  to  public  health. 
These  are  the  txxo  major  xxavs  in  xx  hich  micro- 
biology has  been  useful. 

Historically,  most  efforts  haxe  been  devoted 
to  improx  ing  the  ability  to  control  the  harmful 
effects  of  micro-organisms.  The  industry  recog- 
nized the  extreme  heat  resistance  of  bacterial 
spores  in  the  early  2()th  century  and  sponsored 
or  conducted  much  of  the  early  research  on  the 
mechanisms  of  bacterial  spore  heat  resistance. 
Efforts  to  exploit  the  beneficial  characteristics 
of  micro-organisms,  on  the  other  hand,  have 
been  largely  through  trial-and-error.  Strains 
that  improxe  the  quality  or  character  of  food 
generally  have  been  found,  rather  than  de- 
signed. 


The  idea  of  using  SCP  as  animal  feed  or 
human  food  is  not  nexv;  yeast  has  been  used  as 
food  protein  since  the  beginning  of  the  century. 
How'ex  er,  in  the  past  15  years,  there  has  been  a 
dramatic  increase  in  research  on  SCP  and  in  the 
construction  of  large-scale  plants  for  its  pro- 
duction, especially  for  the  production  of  yeast. 
(See  table  20.)  Interest  in  this  material  is  re- 
flected in  the  numerous  national  and  interna- 
tional conferences  on  SCP,  the  increasing 
number  of  proceedings  and  reviews  published, 
and  the  number  of  patents  issued  in  recent 
years.  (See  table  21.) 

The  issues  addressed  have  covered  topics 
such  as  the  economic  and  technological  factors 
influencing  SCP  processes,  nutrition  and  safety, 
and  SCP  applications  to  human  or  animal  foods. 
Thus  far,  commercial  use  has  been  limited  by 


107 


108  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Table  20.— Estimated  Annual  Yeast  Production,  1977 
(dry  tonnes) 


Baker’s  yeast 

Dried  yeast' 

Europe 

74,000b 

160,000b 

North  America 

73,000 

53,000 

The  Orient 

15,000 

25,000 

United  Kingdom 

15,500 

n 

South  America 

7,500 

2,000 

Africa 

2,700 

2,500 

Totals 187,700  242,500 


®Dried  yeast  includes  food  and  fodder  yeasts:  data  for  petroleum-grown  yeasts 
are  not  available. 

*i*Production  figures  for  U.S.S.R.  not  reported. 

'-None  reported. 

SOURCE:  H.  J.  Peppier  and  D.  Perlman  (eds.),  Microbial  Technoiogy,  vol.  1 (Lo,’- 
don:  Academic  Press,  1979),  p.  159. 


Table  21.— Classification  of  Yeast-Related 
U.S.  Patents  (1970  to  July  1977) 


Category  Number  issued 

Yeast  technology  (apparatus,  processing)  ....  22 

Growth  on  hydrocarbons 28 

Growth  on  alcohols,  acids,  wastes 22 

Production  of  chemicals 14 

Use  of  baking  and  pasta  products 24 

Condiments  and  flavor  enhancers 18 

Reduced  RNA 11 

Yeast  modification  of  food  products 13 

Isolated  protein 5 

Texturized  yeast  protein 7 

Lysates  and  ruptured  cells 7 

Animal  feed  supplements 12 

Total 183 


SOURCE;  H.  J.  Peppier,  “Yeast,”  Annual  Report  on  Fermentation  Processes,  D. 

Perlman  (ed.),  vol.  2 (London:  Academic  Press,  1978),  pp.  191-200. 

several  factors.  For  each  bacterial,  yeast,  or 
algal  strain  used,  technological  problems  (from 
the  choice  of  micro-organisms  to  the  use  of  cor- 
responding raw  material)  and  logistical  prob- 
lems of  construction  and  location  of  plants  have 
arisen.  But  the  primary  limitation  so  far  has 
been  the  cost  of  production  compared  with  the 
costs  of  competing  sources  of  protein.  (The  com- 
parative price  ranges  in  1979  for  selected 
microbial,  plant,  and  animal  protein  products 
are  shown  in  table  22.) 


Table  22.— Comparison  of  Selling  Price  Ranges 
for  Selected  Microbial,  Plant,  and  Animal 
Protein  Products 


Product,  substrate,  and  quality 

Crude 

protein 

content 

Price  range 
1979  U.S. 
dollars/kg 

Single-cell  proteins 

Candida  utilis,  ethanol,  food  grade 

52 

1.32-1.35 

Kluyveromyces  fragilis,  cheese 
whey,  food  grade 

54 

1.32 

Saccharomyces  cerevisiae: 

Brewer’s,  debittered,  food  grade 

52 

1.00-1.20 

Feed  grade 

52 

0.39-0.50 

Plant  proteins 

Alfalfa  (dehydrated) 

17 

0.12-0.13 

Soybean  meal,  defatted 

49 

0.20-0.22 

Soy  protein  concentrate 

70-72 

0.90-1.14 

Soy  protein  isolate 

90-92 

1.96-2.20 

Animal  proteins 

Fishmeal  (Peruvian) 

65 

0.41  -0.45 

Meat  and  bonemeal 

50 

0.24-0.25 

Dry  skim  milk 

37 

0.88-1.00 

SOURCE:  Office  of  Technology  Assessment. 


Agriculture  (USDA),  total  domestic  and  e.xport 
supply  for  U.S.  soybeans  will  gi'ow  7.1  pcM’cent 
by  1985. 

Soybeans  are  primarily  consumed  as  animal 
feed.  But  while  only  4 pei'cent  of  their  annual 
production  are  directly  consumed  by  humans, 
the  market  is  growing  significantly.  The  in- 
troduction of  improved  te.xtured  soy  proU'in  in 
cereals,  in  meat  substitutes  and  e.xtendei's,  and 
in  dairy  substitutes  has  incrc^a.sed  the  us('  of  .soy 
products.  Nevertheless,  the  markc't  does  not  de- 
mand soy  products  in  particular’  hut  pi’otein 
supplements,  vegetable  oils,  feed  gr  ain  supple- 
ments, and  meat  extendcM's  in  gcMKM’al.  Otlier 
protein  and  oil  sources  could  r-eplace  sox  heans 
if  the  economics  were  attr’actii'e  enough.  Fish- 
meal, di'v  beans,  SCP,  and  cer’eals  ar-e  all  jioten- 
tial  competitor's.  As  long  as  a suhstilule  can 
meet  the  nutritional,  fla\'or',  toxicity,  and  r’cgula- 
tory  standards,  competition  will  h(>  pr'imar’ily 
based  on  price. 


The  costs  of  manufacturing  SCP  for  animal 
feed  in  the  United  States  are  high,  particularly 
relative  to  its  major  competing  protein  source, 
soybeans,  which  can  be  produced  with  little  fer- 
tilizer and  minimal  processing.  The  easy  avail- 
ability of  this  legume  severely  limits  microbial 
SCP  production  for  animal  feed  or  human  food. 
In  fact,  according  to  the  U.S.  Department  of 


The  competition  between  .soylx'ans  and  S(  P 
illustrates  one  ol  the  par'adoxes  of  genetic  engi- 
neering. While  signifi('ant  rr'sear’ch  is  attempt- 
ing the  genetic  impr’o\'ement  of  .soybeans,  ge- 
netic techniques  ar'e  also  lu'irig  explor-ed  to  in- 
crease the  production  of  SCI’.  C()nse(|uently,  the 
same  tool— genetic  engiiK'eiing— encourages 
competition  between  the  two  commoditii's 


Ch.6 — The  Food  Processing  Industry  • 109 


Genetic  engineering  and 
SCP  production 

Despite  the  miei'ohial  screening  studies  that 
ha\e  been  conducted  and  the  wealth  of  basic 
genetic  knowledge  a\ailahle  about  common 
\east  (a  majoi'  source  of  SCP),  genetic  engineei'- 
ing  has  had  little  economic  impact  on  S(d’  proc- 
esses until  recently.  Today,  a \ariety  of  sub- 
stances are  being  considered  as  raw  materials 
for  con\ersion. 

• Petruleum-bnsrd  hydrocurbons.—ViuW  re- 
centh’,  the  w ide  a\  ailahility  and  low  cost  of 
peti'ochemicals  ha\  e made  the  /j-alkane  hy- 
drocarbons (straight  chain  molecules  of 
carbon  and  hydrogen),  which  are  petro- 
chemical by{)roducts,  potential  raw  materi- 
als for  SCd’  production.  .At  British  Petro- 
leum, mutants  of  micro-organisms  ha\e 
been  obtained  ha\  ing  an  increased  protein 
content.  .Mutants  ha\e  also  been  found 
with  other  increased  nutritive  \ alues,  e.g., 
vitamin  content. 

• Methane  or  met/tano/.— Relatively  few  ge- 
netic studies  have  been  directed  at  in\  esti- 
gating  the  genetic  control  of  the  microbial 
use  of  methane  or  methanol.  However,  one 
recent  application  of  genetic  engineering 
has  been  reported  bv  the  Imperial  Chem- 
ical Industries  (ICI)  in  the  United  Kingdom, 
where  the  genetic  makeup  of  a bacterium 
{Methylophilus  methylotrophus)  has  been 
altered  so  that  the  organism  can  grow 
more  readily  on  methanol.  The  increase  in 
growth  pro\  ides  increased  protein  and  has 
made  its  production  less  expensive.  The 
genetic  alteration  was  accomplished  by 
transferring  a gene  from  Escherichia  coli  to 
M.  methylotrophus. 

• Carbohydrates.— Many  carbohydrate  sub- 
strates—from  starch  and  cellulose  to  beets 
and  papermill  wastes— have  been  investi- 
gated. Forests  are  the  most  abundant 
source  of  carbohydrate  in  the  form  of  cel- 
lulose. But  before  it  can  be  used  by  micro- 
organisms, it  must  be  transformed  into  the 
carbohydrate,  glucose,  by  chemical  or  en- 
zymatic pretreatment.  Many  of  the  SCP 
processes  that  use  cellulose  employ  orga- 


nisms that  produce  the  enzyme  cellulase, 
w hich  degrades  cellulose  to  glucose. 

Most  of  the  significant  genetic  studies  on  the 
pi'oduction  of  cellulase  by  micro-organisms  are 
just  beginning  to  appear  in  the  literature.  I'he 
most  recent  experiments  have  been  successful 
in  ci'eating  fungal  mutants  that  produce  excess 
amounts. 

Commercial  protluction 

Of  the  estimated  2 million  tons  of  SCd’  pro- 
duced annually  thi'oughout  the  w'orld,  most 
comes  from  cane  and  beet  molasses,  w'ith  about 
oOO, ()()()  tons  from  hydi'olyzed  wood  wastes, 
corn  trash,  and  papermill  wastes.  (See  table  23.) 

Integrated  systems  can  he  designed  to  couple 
the  production  of  a product  oi-  food  with  SCP 
production  from  wastes.  E.g.,  the  waste  saw- 
dust from  the  lumber  industry  could  become  a 
source  of  cellulose  for  micro-organisms.  Id’s 
successful  genetic  engineering  of  a micro-orga- 
nism to  increase  the  usefulness  of  one  raw 
material  (methanol)  should  encourage  similar 
attempts  for  other  raw  materials. 

But  while  SCP  can  he  obtained  from  a wide 
variety  of  micro-organisms  and  raw'  materials, 
the  nutritional  value  and  the  safety  of  each 
micro-organism  vary  widely,  as  do  the  costs  of 
competing  protein  sources  in  regional  markets. 
Consequently,  accurate  predictions  cannot  be 
made  about  the  likelihood  that  SCP  will  displace 
traditional  protein  products,  overall.  Displace- 
ments have  and  will  continue  to  occur  on  a case- 
by-case  basis. 


Table  23.— Raw  Materials  Already  Tested  on  a 
Laboratory  or  Small  Plant  Scale 


Agave  juices 

Pulpmill  wastes 

Barley  straw 

Sawdust 

Cassava 

Sunflower  seed  husks 

Citrus  wastes 

(treated) 

Date  carbohydrates 

Wastes  from  chemical 

IVleatpacking  wastes 

production  of  maleic 

IVIesquite  wood 

anhydride 

Peat  (treated) 

Waste  polyethylene  (treated) 

SOURCE:  Office  of  Technology  Assessment. 

110  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


Genetics  in  baking;  brewing;  and  winemaking 


The  micro-organism  of  greatest  significance 
in  the  baking,  brewing,  and  winemaking  indus- 
tries is  common  yeast.  Because  of  its  impor- 
tance, yeast  was  one  of  the  first  micro-orga- 
nisms to  be  used  in  genetic  research.  Neverthe- 
less, the  surge  in  studies  in  yeast  genetics  has 
not  been  accompanied  by  an  increase  in  its 
practical  application,  for  three  reasons: 

• industries  already  have  the  desired  effi- 
cient strains,  mainly  as  a result  of  trial-and- 
error  studies; 

• new  genetic  strains  are  not  easily  bred; 
they  are  incompatible  for  mating  and  their 
genetic  characteristics  are  poorly  under- 
stood; and 

• many  of  the  important  characteristics  of 
industrial  microbes  are  complex;  several 
genes  being  responsible  for  each. 

Changing  technologies  in  the  brewing  indus- 
try and  increased  sophistication  in  the  molec- 
ular genetics  of  yeast  have  made  it  possible  for 
researchers  to  achieve  novel  goals  in  yeast 
breeding.  One  strain  that  has  already  been  con- 
structed can  produce  a low-carbohydrate  beer 
suitable  for  diabetics.  (See  figure  26.) 

The  baking  industry  is  also  undergoing  tech- 
nological revolution,  and  yeasts  with  new  prop- 
erties are  now  needed  for  the  faster  fermenta- 
tion of  dough.  New  strains  with  improved  bio- 
logical activity,  storage  stability,  and  yield 
would  allow  improvements  in  the  baking  proc- 
ess. 

In  the  past,  most  genetic  applications  have 
come  in  the  formation  of  hybrid  yeasts.  The 
newer  genetic  approaches,  which  use  cell  fu- 
sion now  open  up  the  possibility  of  hybrids  de- 
veloped from  strains  of  yeast  that  carry  useful 
genes  but  cannot  mate  normally. 

Classical  genetic  research  has  also  been  car- 


Figure  26.— The  Use  of  Hybridization  To  Obtain 
a Yeast  Strain  for  the  Production  of 
Low-Carbohydrate  Beer 


Saccharomyces 

Saccharomyces 

carisbergensis  (yeast) 

Mated  with 

diasticus  (yeast) 

(strain  1)a 

(dextrin  fermenting 
ability) 

Hybrid  yeast  1 

Mated  with 

Saccharomyces 

(palatabie  beer) 

carisbergensis 
(strain  1) 

Hybrid  yeast  ii 
(palatabie  beer) 

Mated  with 

Wild  yeast 
(isomaltose, 
isomaltotriose 
fermenting  ability) 

Hybrid  yeast  III 
(produces  diabetic 
beer) 

^Strain  1 1s  a brewing  yeast. 

SOURCE:  Office  of  Technology  Assessment. 


ried  out  with  wincf  v(fasts.  Interestingly,  within 
the  past  10  years,  scientists  haw  isolated  in- 
duced mutants  of  witK*  yeasts  that  haw;  II  an 
increased  alcohol  tolerance  and  the  ea|)aeit\'  to 
completely  ferment  grapt'  extracts  of  unusually 
high  sugar  contcfiit;  2)  impro\(*d  sedimentation 
properties,  im|)ro\  ing  or  facilitating  separation 
of  yeasts  from  the  w incf;  and  ;0  improwd  per- 
formance in  the  production  of  certain  types  of 
wines.  Hybridization  studies  of  wine  yeasts 
have  been  actixely  pursiu'd  only  recently. 

Progress  in  de\(’loping  strains  of  y(>ast  w ith 
novel  properticfs  is  limitc'd  by  the  lack  of  (Miough 
suitable  approxcnl  systems  lor  using  recombi- 
nant DNA  (rl)N/\)  tcM'hnology.  I.vcntual  approv- 
al by  the  Kcfcomhinant  l)N  \ \d\  i.sory  ( ommit- 
tee  is  ex[)(fct(ul  to  boost  applied  research  lor  the 
Ijrewing,  baking,  and  w inemaking  industries 


I 


Ch.  6— The  Food  Processing  Industry  • 111 


\ 

Microbial  polysaccharides 

rhe  food  [)i’ot’essin^  industry  uses  [jolysac- 
eharides  (poK  iiierie  sugars)  to  alter  or  contiol 
the  physical  pi'operties  of  foods.  Many  ai'e  in- 
corporated into  foods  as  tliickeners,  gelling 
; agents,  and  agents  to  control  ice  crystal  foi  ina- 
' tion  in  frozen  foods.  They  are  used  in  instant 
foods,  salad  dressings,  sauces,  whips,  to[)pings, 
processed  cheeses,  and  tlair\'  products.  New 
uses  are  constantly  appearing.  The  annual  mai'- 
ket  in  the  I'nited  States  is  leported  to  he  o\er 
36,000  tons,  not  including  starches  atid  deri\  a- 
ti\  es  of  cellulose. 

Since  many  of  the  pol\ meric  sugars  now  used 
in  food  processing  are  derixed  from  plant 
sources,  microbial  polysaccharides  ha\e  had 
limited  use.  To  compete  economically,  a micro- 
bial pohsaccharide  must  offer  new  properties, 
meet  all  safety  requirements,  and  he  readily 
av  ailable.  \ ery  few  have  reached  the  level  of 
commercial  applications:  the  onlv  one  in  large- 
scale  commercial  production  is  .xanthan  gum.* 


'The  history  of  the  development  of  .xanthan  gum  indicates  that 
the  commercially  significant  organisms  resulted  from  an  extensive 
screening  program  for  gum  producers  stored  in  the  .Northern  Uti- 


,\  wide  variety  of  polysaccharides  could  theo- 
retically he  produced  foi’  use  in  food  processing. 
.Applied  genetics  may  increase  their  production, 
modify  those  that  are  produced,  eliminate  the 
degi'adative  enzymes  that  break  them  down,  or 
change  the  microbes  that  produce  them.  How- 
ever, as  with  other  microbial  processes,  the  ap- 
plication of  genetics  depends  on  an  understand- 
ing of  both  the  biochemical  pathway  for  synthe- 
sis of  a given  polysacchai'ide  and  the  systems 
that  control  microbial  production.  For  many  mi- 
crobial polysaccharides,  this  information  does 
not  vet  exist:  furthermore,  little  is  known  about 
the  enzymes  that  may  he  used  to  modify  poly- 
saccharides to  more  useful  forms.  Progress  will 
only  he  able  to  occur  when  these  information 
gaps  are  filled. 

lii'alion  Kcscai'i'h  and  Ocvclopmcnl  Division  of  llSO.V's  largo  mi- 
(■|■ol)ial  fullurc  colleclion.  Xanllian  gum  produced  hv  Xanlhomo- 
nas  camppstris  \RRI.  R- 14.)9  was  found  lo  ha\  e characlerislics  that 
rendei  ed  it  \ei'v  promising  as  a commercial  product.  In  19G0,  the 
Kelco  ilivisioti  of  VIerck  <4 Co..  Inc.,  cai  ried  out  pilot  plant  feasihili- 
ty  stiulies.  and  suhstantial  commercial  pi'oduction  began  in  19(i4. 
Vllhough  much  of  the  work  to  date  has  hee?i  (uirried  out  with 
polysaccharides  from  one  particular  strain,  there  is  increasing  e\  i- 
dence  to  suggest  that  they  could  also  he  produced  fj'om  other 
strains. 


Enzymes  

Enzymes  are  produced  for  industrial,  med- 
ical, and  laboratory  use  both  by  fermentation 
processes  that  employ  bacteria,  molds,  and 
yeasts  and  by  extraction  from  natural  tissues. 
The  present  world  market  for  industrial  en- 
zymes is  estimated  to  be  S150  million  to  SI 74 
million:  the  technical  (laboratory)  market  adds 
another  S20  million  to  S40  million.  Fewer  than 
50  microbial  enzymes  are  of  industrial  impor- 
tance today,  but  patents  have  been  granted  for 
more  than  a thousand.  This  reflects  the  increas- 
ing interest  in  developing  new  enzyme  prod- 


ucts; it  also  show's  that  it  is  easier  to  discover  a 
new  enzyme  than  to  create  a profitable  applica- 
tion for  it.* 

Most  industrial  enzymes  are  used  in  the  de- 
tergent industry  and  the  food  processing  in- 

*The  enzvme  literature  is  exlensiv'e  aiid  comprises  well  over 
10,000  papers  per  year.  .Although  less  than  SO  percent  of  these 
publications  are  concerned  with  microbial  enzymes  and  most  are 
found  to  have  no  industrial  interest,  a few  thousand  papers  per 
year  are  of  potential  interest  for  the  industrial  development  of  en- 
zymes. Less  than  100  papers  dealing  with  industrial  processes  ap- 
pear e\  erv  year,  and  few  descrjbe  processes  of  great  economic  sig- 
nificance. 


112  • Impacts  of  Applied  Genetics— Micro-Organisms,  Piants,  and  Animals 


dustry,  particularly  for  starch  processing.  En- 
zymes began  to  be  used  in  quantity  only  20 
years  ago.  In  the  early  1960’s,  glucoamylase  en- 
zyme treatment  began  to  replace  traditional 
acid  treatment  in  processing  starch;  around 
1965,  a stable  protease  (an  enzyme)  was  in- 
troduced into  detergent  preparations  to  help 
break  down  certain  stains;  and  in  the  1970’s, 
glucose  isomerase  was  used  to  convert  glucose 
to  fructose,  practically  creating  the  high-fruc- 
tose corn  syrup  industry. 

Genetic  engineering  and  enzymes  in 
the  food  processing  industry 

Biotechnology  applied  to  fermentation  proc- 
esses will  make  available  larger  quantities  of  ex- 
isting enzymes  as  well  as  new  ones.  (See  ch.  5.) 
The  role  of  genetic  engineering  in  opening  com- 
mercial possibilities  in  the  food  processing  in- 
dustry is  illustrated  by  the  enzyme,  pullulanase. 
This  enzyme  degrades  pullulan,  a polysaccha- 
ride, to  the  maltose  or  high-maltose  syrups  that 
give  jams  and  jellies  improved  color  and  bril- 
liance. They  reduce  off-color  development  pro- 
duced by  heat  in  candies  and  prevent  sandiness 
in  ice  cream  by  inhibiting  sugar  crystallization. 
Maltose  has  several  unique  and  favorable  char- 
acteristics. It  is  the  least  water-absorbent  of  the 
maltose  sugars  and,  although  it  is  not  as  sweet 
as  glucose,  it  has  a more  acceptable  taste.  It  is 
also  fermentable,  nonviscous,  and  easily  solu- 
ble. It  does  not  readily  crystallize  and  gives  de- 
sirable browning  reactions. 

Pullulanase  can  also  break  down  another  car- 
bohydrate, amylopectin,  to  produce  high  amy- 
lose  starches.  These  starches  are  used  in  indus- 
try as  quick-setting,  structurally  stable  gels,  as 
binders  for  strong  transparent  films,  and  as 
coatings.  Their  acetate  derivatives  are  added  to 
textile  finishes,  sizing,  adhesives,  and  binders. 
In  food,  amylose  starches  thicken  and  give  tex- 
ture to  gumdrop  candies  and  sauces,  reduce  fat 
and  grease  in  fried  foods,  and  stabilize  the  pro- 
tein, nutrients,  colors,  and  flavors  in  reconsti- 
tuted products  like  meat  analogs. 

In  view  of  the  current  shortages  of  petro- 
leum-derived plastics  and  the  need  for  a biode- 
gradable replacement,  amylose’s  ability  to  form 


plastic-like  wraps  may  prox  ide  its  largest  indus- 
trial market,  although  that  market  has  not  yet 
been  dex  eloped. 

If  applications  for  the  products  made  l)v 
pullulanase  can  be  dex  eloped,  genetic  engineei'- 
ing  can  he  used  to  insert  this  enzyme  into  in- 
dustrially useful  organisms  and  to  increase  its 
production.  Howexer,  the  food  processing  in- 
dustry is  permitted  to  use  only  enzymes  that  are 
obtained  from  sources  approxed  for  food  use. 
Since  the  chief  source  of  pullulanase  is  a patho- 
genic bacterium,  Klebsiella  aerogenes,  no  signifi- 
cant efforts  hax  e been  made  to  apply  genetics  to 
improve  its  production  or  (lualitv.  Molecular 
genetics  could  ultimately  transfer  the  pullula- 
nase trait  from  K.  aerogenes  to  a micro-organism 
approved  for  food  use,  if  a])prox  ed  micro-oi'ga- 
nisms  that  manufacture  pullulanase  cannot  he 
found. 

Sweeteners,  flavors,  and  fragrances 

Biotechnology  has  already  had  a markc'd  im- 
pact on  the  sxveetener  industry.  I he  ax  ailabilitx 
of  the  enzymes  glucose  isonu'rase,  inxcrta.se, 
and  amylase  has  made  the  production  of  high- 
fructose  corn  sweetenei's  (III'(’S)  pi'ofitahle.  Pro- 
duction of  HFCS  in  the  Unitcul  Stat('s  has  in- 
creased from  x'irtually  nothing  in  1970  to  10 
percent  of  the  entire  productit)n  ol  ('alori(' 
sxveeteners  in  1980  (11  Ih  p('r  capita).  TIk*  price 
advantage  of  HFCS  is  expcctc'd  to  cause  its  con- 
tinued groxvth,  particularly  in  the  hcx('ragc  in- 
dustry. In  fact,  the  (^oca  (’ola  ( o.  announced  in 
1980  that  fructose  will  .soon  constitute  as  much 
as  50  percent  of  the  sxvecteiK'i’  u.sc'd  in  its  name 
brand  bex  erage. 

Biotechnology  can  b(?  us(’d  to  product*  other 
sxveeteners  as  well.  While  it  is  unlikt'lx  that  su- 
crose xvill  ever  be  mad(?  In  micro-organisms  (al- 
though impi’oxements  in  sugarcane  and  sugar 
beet  yields  may  result  fi'om  agricultural  genetic 
studies,  see  ch.  8 ),  th(*  microbial  production  ol 
loxv-caloric  sxxeeteiKM's  is  a distinct  possibility 
Three  nexv  ex|)erimental  sxx  (’cteners— as|)ai - 
tame,  monellin,  and  thaumatin— arc  candidates 

Aspartame  is  synthesized  cln'micallx  Irnm 
the  amino  acids,  aspartic  acid  and  phcnxiala- 
nine,  which  can  thtMii.selx vs  he  madt*  In  Icnncn- 


Ch.  6— The  Food  Processing  Industry  • 113 


tation.  riie  possibilit\  of  using  microbes  to  cou- 
[)le  tbe  two  amino  acids  is  being  imestigated  in 
at  least  one  biotecbnoIog\'  I’esearcb  fii  in.  Cliem- 
I ical  production  of  as|Kirtame  is  e.\pensi\e  and 
benefits  from  biotecbnologx'  are  possible. 

Monellin  and  thaumatin  are  natural  sub- 
stances—proteins  obtained  from  W est  African 
plants.  Both  are  intenseK’  sweet— up  to  100, ()()() 
times  sweeter  than  table  sugar— and  the  sensa- 
tion of  sweetness  can  last  for  hours.  Their 
microbial  pi'oduction  ma\  be  competiti\e  with 
tbeii’  e.xtraction  from  plants.  Since  the  physical 
and  biological  properties  of  thaumatin  are 
known,  it  might  also  be  prcKluced  through  ge- 
netic engineering.  Such  an  approach  would  not 
onl\'  increase  the  available  su[)ply,  but  would 
offer  new  molecules  for  in\  estigating  tbe  phvsi- 
olog\'  of  taste. 


Other  flavors  and  fragrances  show  less  prom- 
ise at  present.  Although  tbe  chemistry  of  sev- 
eral flav  ors  and  aromas  has  been  identified,  too 
little  research  into  their  use  has  been  con- 
ducted. * 


'Ret't'iil  woi'k  on  tin’  Ibrmation  by  niic'm-oi'gani.snis  of  flavor 
and  aroma  chcmiral.s  known  as  larlones  and  toi'penoids  has  been 
roporlcd.  I^iclones  ociiir  as  flavor-contributinf'  components  in 
main  fermentation  products,  w here  they  are  formed  by  microltial 
reactions.  Different  |)alhways  e.xist  for  their  microbial  foi  niation. 
I■.(^.,  {'amma-luityrolactone,  which  is  formed  diii'ing  yeast  fermen- 
tation. is  found  in  sherry,  wine,  and  beer.  As  early  as  1930,  an  or- 
ganism was  i.solated  from  orange  lea\  es  that  had  a peach-like  odor 
and  was  thought  to  he  Sporoholomyces  roseus.  The  lactones,  4- 
di'canolide  and  cis-6-dodecen-4-olide  were  found  to  he  responsi- 
ble. 


Overview  

The  application  of  genetic  engineering  will  af- 
fect the  food  processing  industry  in  piecemeal 
fashion.  Isolated  successes  can  be  e.xpected  for 
certain  food  additives,  such  as  aspartame  (not 
yet  approved  hv  the  Food  and  Drug  Administra- 
tion (FDA)  for  sale  in  the  United  States)  and  fruc- 
tose, and  for  improvements  in  SCP  production. 
But  an  industrywide  impact  is  not  expected  in 
the  near  future  because  of  several  conflicting 
forces: 

• The  basic  genetic  knowledge  of  character- 
istics that  could  improve  food  has  not  been 
adequately  dev  eloped. 

• The  food  processing  industry  is  conserva- 
tive in  its  research  and  development  ex- 
penditures for  improved  processes,  gener- 
ally allocating  less  than  half  as  much  as 
more  technologically  sophisticated  indus- 
tries. 

• Products  made  by  new  microbial  sources 
must  satisfy  FDA  safety  regulations,  which 
include  undergoing  tests  to  prove  lack  of 
harmful  effects.*  It  may  be  possible  to  re- 

*E.g.. all  food  additi\es  and  micro-organisms  used  in  food  proc- 
essing must  be  approved  as  generally  regarded  as  safe. 


duce  the  amount  of  required  testing  by 
transferring  the  desired  gene  into  micro- 
organisms that  already  meet  FDA  stand- 
ards. 

Nevertheless,  the  application  of  new  genetic 
technologies  will  probably  accelerate.  Techno- 
logically sophisticated  companies  are  being 
drawn  into  the  business.  Traditionally  capital- 
intensive  companies  such  as  Union  Carbide, 
ITT,  General  Electric,  Corning  Glass,  and 
McDonnell-Douglas  can  be  expected  to  intro- 
duce automation  and  more  sophisticated  engi- 
neering to  food  processing,  modernizing  the  in- 
dustry’s technology.  As  has  been  noted  by  one 
industry  observer:* 

You  don’t  work  on  a better  way  to  preserve 
fish.  You  try  to  change  the  system  so  that  you 
no  longer  catch  fish;  you  "manufacture”  them 
and,  if  possible,  do  it  right  on  top  of  your  mar- 
ket so  that  you  don’t  have  to  preserve  them  at 
all. 


'M.  L.  Kastens,  "The  Coming  Food  Industry,"  Chemtech,  April 
1980,  pp.  215-217. 


114  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


You  don’t  worry  about  processing  bacon 
without  nitrites,  you  engineer  a synthetic  bacon 
with  designed-in  shelf  life. 

You  don’t  try  to  educate  people  to  eat  a "bal- 
anced diet;’’  you  create  a "whole”  food  with  the 
proper  balance  of  nutrients  and  supplements, 
and  you  make  it  taste  like  something  people 
already  like  to  eat. 


Genetic  engineering  can  be  expected  to  aid  in 
the  creation  of  novel  food  preparations  through 
effects  on  both  the  food  itself  and  the  additives 
used  for  texturizing,  flavoring,  and  preserving. 


chapter  7 

The  Use  of  Genetically 
Engineered  Micro-Organisms 

in  the  Environment 


chapter  7 


Page 


Mineral  Leaching  and  Recovery 117 

Microbial  Leaching 117 

Applied  Genetics  in  Strain  Improvement 118 

Metal  Recovery 118 

Oil  Recovery 119 

Enhanced  Oil  Recovery 119 

Microbial  Production  of  Chemicals  Used 

in  EOR 120 

In  Situ  Use  of  Micro-Organisms 121 

EOR  and  Genetic  Engineering 122 

Constraints  to  Applying  Genetic 

Engineering  Technologies  in  EOR 122 

Genetic  Engineering  of  Micro-Organisms 
for  Use  in  Other  Aspects  of  Oil  Recovery 

and  Treatment 123 

Overview  of  Genetic  Engineering  in  Mining 

and  Oil  Recovery 123 


Page 

Pollution  Control 123 

Enhancing  Existing  Microbial  Degradation 

Activity 124 

Adding  Microbes  to  Clean  Up  Pollution 124 

Commercial  Applications— Market  Size  and 

Prospects 125 

Genetic  Research  in  Pollution  Control 126 

Federal  Research  Support  for  Engineering 
Microbes  to  Detoxify  Hazardous  Substances  . 127 

Summary 127 

Issue  and  Options— Biotechnology 128 


Figure 


Figure  No.  Page 

27.  Chemical  Flooding  Process 120 


chapter  7 

The  Use  of  Genetically  Engineered 
Micro-Organisms  in  the  Environment 


Although  most  genetically  engineered  micro- 
organisms are  being  designed  for  contained  fa- 
cilities like  fermenters,  some  are  being  exam- 
ined for  their  usefulness  in  the  open  en\  iron- 
ment  for  such  purposes  as  mineral  leaching  and 
reco\  erv,  oil  reco\  erv,  and  pollution  control. 

■All  three  applications  are  characterized  by: 

• the  use  of  large  \olumes  of  micro-orga- 
nisms: 


• less  control  o\er  the  behavior  and  fate  of 
the  micro-organisms; 

• a possibility  of  ecological  disruption;  and 

• less  basic  research  and  development  (R&.D) 
—and  a higher  degree  of  speculation— than 
the  industries  previously  discussed. 


Mineral  leaching  and  recovery 


.All  micro-organisms  interact  with  metals. 
Two  interactions  that  are  of  potential  economic 
and  industrial  interest  are  leaching  metals  from 
their  ores,  and  concentrating  metals  from 
wastes  or  dilute  mixtures.  The  first  would  allow 
the  extraction  of  metals  from  large  quantities  of 
low-grade  ores:  the  second  would  provide  meth- 
ods for  recycling  precious  metals  and  control- 
ling pollution  caused  by  toxic  metals. 

Microbial  leaching 

In  microbial  or  bacterial  leaching,  metals  in 
ores  are  made  soluble  by  bacterial  action.  Even 
before  bacterial  leaching  systems  became  ac- 
cepted industrial  practice,  it  was  known  that 
dissolved  metals  could  be  recovered  from  mine 
and  coal  wastes.  Active  mining  operations  cur- 
rently based  on  this  process  (such  as  those  in 
Rio  Tinto,  Spain)  date  back  to  the  18th  century. 
Presently,  large-scale  operations  in  the  United 
States  use  bacterial  leaching  to  recover  copper 
from  waste  material.  Estimates  for  the  contri- 
bution of  copper  leaching  to  the  total  annual 
U.S.  production  range  from  11.5  to  15  percent. 

Leaching  begins  with  the  circulation  of  water 
through  large  quantities— often  hundreds  of 
tons— of  ore.  Bacteria,  which  are  naturally  asso- 
ciated with  the  rocks,  then  cause  the  metals  to 


be  leached  by  one  of  two  general  mechanisms: 
either  the  bacteria  act  directly  on  the  ore  to  ex- 
tract the  metal  or  they  produce  substances, 
such  as  ferric  iron  and  sulfuric  acid,  which  then 
extract  the  metal.  It  appears  that  simply  adding 
acid  is  not  as  efficient  as  using  live  bacteria. 
Although  acid  certainly  plays  a role  in  metal  ex- 
traction, it  is  possible  that  direct  bacterial  attack 
on  some  ores  is  also  involved.  In  fact,  some  of 
the  bacteria  that  are  known  to  be  involved  in 
mineral  leaching  have  been  shown  to  bind  tena- 
ciously to  those  minerals. 

The  application  of  the  leaching  process  to 
uranium  mining  is  of  particular  interest  be- 
cause of  the  possibility  of  in  situ  mining.  Instead 
of  using  conventional  techniques  to  haul  urani- 
um ore  to  the  surface,  microbial  suspensions 
can  extract  the  metal  from  its  geological  setting. 
Water  is  percolated  through  underground 
shafts  where  the  bacteria  dissolve  the  metals. 
The  solution  is  then  pumped  to  the  surface 
where  the  metal  is  recovered.  This  approach, 
also  called  "underground  solution  mining,”  is 
already  used  in  Canadian  uranium  mines, 
where  it  began  almost  by  chance.  In  1960,  after 
only  2 years  of  operation,  researchers  at  the 
Stanrock  Uranium  Mine  found  that  the  natural 
underground  water  contained  large  amounts  of 
leached  uranium.  In  1962,  over  13,000  kilo- 


777 


118  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


grams  (kg)  of  uranium  oxide  were  obtained 
from  the  water.  Thereafter,  water  was  circu- 
lated through  the  mines  as  part  of  the  mining 
operation.  It  has  been  suggested  that  extending 
this  practice  to  most  mines  would  have  signifi- 
cant environmental  benefits  because  of  the 
minimal  disruption  of  the  land  surface. 
Although  the  process  is  slower  than  the  technol- 
ogy currently  employed,  the  operating  costs 
might  be  lower  because  of  the  simplicity  of  the 
system,  since  no  grinding  machinery  is  needed. 
Furthermore,  deeper  and  lower  grade  deposits 
could  be  mined  more  readily. 

Bacterial  leaching  can  also  extract  sulfur-con- 
taining compounds,  such  as  pyrite,  from  coal, 
producing  coal  with  a lower  sulfur  content. 
Sulfur-containing  coals  from  such  areas  as  Ohio 
and  the  Appalachian  Mountains  are  now  less  de- 
sirable than  other  coals  because  of  the  sulfur 
dioxide  they  release  during  burning.  They  often 
contain  up  to  6 percent  sulfur,  of  which  70  per- 
cent can  be  in  the  form  of  pyrite.  According  to 
recent  data,  mixed  populations  of  different  bac- 
teria, rather  than  a single  species,  are  respon- 
sible for  the  most  effective  removal  of  sulfur— a 
finding  that  may  lead  to  the  genetic  engineering 
of  a single  sulfur-removing  bacterium  in  the 
future. 

Applied  genetics  in  strain  improvement 

The  bacterium  most  studied  for  its  leaching 
properties  has  been  Thiobacillus  ferroo^idans 
(which  leaches  copper),  but  others  have  also 
been  identified  in  natural  leaching  systems. 
Although  leaching  ability  is  probably  under 
genetic  control  in  these  organisms,  practically 
nothing  is  known  about  the  precise  mecha- 
nisms. This  is  largely  because  little  information 
exists  in  two  critical  areas:  the  chemistry  of  in- 
teraction between  the  bacteria  and  rock  sur- 
faces; and  the  genetic  structure  of  the  micro- 
organisms. The  finding  that  mixed  populations 
of  bacteria  interact  to  increase  leaching  efficien- 
cy complicates  the  investigation. 

Because  of  the  lack  of  genetic  and  biochemi- 
cal information  about  these  bacteria,  the  appli- 
cation of  genetic  technologies  to  mineral  leach- 
ing remains  speculative.  Progress  in  obtaining 


more  information  is  slow  because  less  than  a 
dozen  laboratories  in  the  Nation  are  actively 
performing  research. 

But  even  when  the  scientific  knowledge  is 
gathered,  two  obstacles  to  the  use  of  genetically 
engineered  micro-organisms  will  remain.  The 
first  is  the  need  to  develop  engineered  systems 
on  a scale  large  enough  to  exploit  their  biologi- 
cal activities.  A constant  interchange  must  take 
place  between  microbial  geneticists,  geologists, 
chemists,  and  engineers.  E.g.,  the  geneticists 
must  understand  the  needs  identified  by  the 
geologists  as  well  as  the  problems  faced  by  the 
engineers,  who  must  scale-up  laboratory-scale 
processes.  The  complex  nature  of  the  problem 
can  be  approached  most  successfully  by  an 
interdisciplinary  group  that  recognizes  the 
needs  and  limitations  of  each  discipline. 

The  second  obstacle  is  en\ironmental.  In- 
troducing large  numbers  of  genetically  engi- 
neered micro-organisms  into  the  en\ii'onment 
raises  questions  of  possible  ecological  disrup- 
tion, and  liability  if  damage  occurs  to  the  ('ini- 
ronrnent  or  human  health. 

In  summary,  the  present  lack  of  sufficient 
scientific  knowledge,  scientists,  and  interdis- 
ciplinary teams,  and  the  concei'iis  for  ec'ological 
safety  present  the  major  obstacles  to  the  use  of 
genetic  engineering  in  microbial  leaching. 

Metal  recovery 

The  use  of  micro-organisms  to  concentrate* 
metals  from  dilute  solutions  suc'h  as  individual 
waste  streams  has  two  goals:  to  re-cover  metals 
as  part  of  a recycling  process:  and  to  ('liminate* 
any  metal  that  may  lie  a pollutant,  I he  process 
makes  use  of  the  ability  of  micro-organisms  to 
bind  metals  to  their  surfaces  and  then  concen- 
trate them  internally. 

Studies  at  the  Oak  Kidge  National  l.ahoratory 
in  Tennessee  have  shown  that  micio-organisms 
can  he  used  to  remove  heavy  metals  from  indus- 
trial effluents.  Metals  sucli  as  cohalt,  nickel, 
silver,  gold,  uranium,  and  plutonium  in  concen- 
trations of  less  than  1 j)art  |)er  million  (ppm)  can 
be  recovered.  The  process  is  particularly  usetui 
for  recovering  metals  fi'om  dilute  solutions  ol 


c/7.  7— The  Use  of  Genetically  Engineered  Micro-Organisms  in  the  Environment  *119 


10  to  100  ppm,  v\here  nonbiological  methods 
ma\  he  uneconomical.  Organisms  such  as  the 
common  \east  Saccharonn  t'es  cerevisiae  can  ac- 
cumulate uranium  up  to  20  [)ercent  of  their 
total  weight. 

rhe  economic  competitiveness  of  biological 
methods  has  not  yet  been  proven,  hut  genetic 
improvements  have  been  attempted  only  re- 
centlv.  The  cost  of  producing  the  micro-orga- 
nisms has  been  a major  consideration.  If  it  can 
he  reduced,  however,  the  approach  might  he 
useful. 


-As  with  other  biological  systems,  genetic 
engineering  may  increase  the  efficiency  of  the 
extraction  process.  In  the  Saccharomyces  sys- 
tem, differences  in  the  ability  to  recov'er  the 
metals  have  been  demonstrated  within  popula- 
tions of  cells.  Selection  for  cells  with  the  genetic 
ability  to  accumulate  large  amounts  of  specific, 
desired  metals  would  he  an  important  step  in 
designing  a practical  system. 


Oil  recovery  

V 

Since  1970,  oil  production  in  the  Lhiited 
States  has  declined  steadily.  The  supply  can  he 
increased  by:  accelerating  explorations  for  new 
oilfields;  by  mining  oil  shale  and  coal  and  con- 
verting them  to  liquids;  and  by  developing  new 
methods  for  recov  ering  oil  from  existing  reser- 
voirs. 

In  primary  methods  of  oil  recovery,  natural 
expulsive  forces  (such  as  physical  expansion) 
drive  the  oil  out  of  the  formation.  In  secondary 
methods  of  recovery,  a fluid  such  as  water  or 
natural  gas  is  injected  into  the  reservoir  to  force 
the  oil  to  the  well.  .Approximately  50  percent  of 
domestic  crude  in  recent  years  has  been  ob- 
tained through  secondary  recovery. 

Recently,  new  methods  of  oil  recovery  have 
been  added  to  primary  and  secondary  methods, 
which  are  called  tertiary,  improved,  or  en- 
hanced oil  recovery  (EOR)  techniques.  They  em- 
ploy chemical  and  physical  methods  that  in- 
crease the  mobility  of  oil,  making  it  easier  for 
other  forces  to  drive  it  out  of  the  ground.  The 
major  target  for  EOR  is  the  oil  found  in  sand- 
stone and  limestone  formations.  It  is  here  that 
applied  genetics  may  play  a major  role, 
engineering  micro-organisms  to  aid  in  recovery. 

Oil  susceptible  to  these  processes  is  localized 
in  reservoirs  and  pools  at  depths  ranging  from 
100  ft  to  more  than  17,000  ft.  In  these  areas,  the 
oil  is  adsorbed  on  grains  of  rock,  almost  always 
accompanied  by  water  and  natural  gas.  The 


physical  association  of  the  trapped  oil  and  the 
surrounding  geological  formations  varies  signif- 
icatitly  from  site  to  site.  The  unknown  charac- 
teristics of  these  variations  are  largely  respon- 
sible for  the  economic  risk  in  an  attempted  EOR. 

Enhanced  oil  recovery 

Of  the  original  estimated  volume  of  more 
than  450  billion  barrels  (hbl)  of  U.S.  oil  reserves, 
about  120  billion  hbl  have  been  recovered  by 
primary  and  secondary  techniques,  and  another 
30  billion  hbl  are  still  accessible  by  these 
methods.  The  remaining  300  billion  bbl  how- 
ever, are  probably  recoverable  only  by  EOR 
methods.  These  figures  include  the  oil  remain- 
ing in  known  sandstone  and  limestone  reser- 
voirs and  exclude  tar  sands  and  oil  shale. 

Four  EOR  processes  are  currently  used.  All 
are  designed  to  dislodge  the  crude  oil  from  its 
natural  geological  setting: 

• In  thermal  processes,  the  oil  reservoir  is 
heated,  which  causes  the  viscosity  of  the  oil 
to  decrease,  and  with  the  aid  of  the 
pressure  of  the  air  introduced,  supports 
the  combustion  that  forces  the  petroleum 
to  the  producing  well.  Thermal  processes 
will  not  be  improved  by  genetic  technol- 
ogies. 

• Various  crude  oils  differ  in  their  viscosity- 
ability  to  flow.  Primary  and  secondary 
methods  can  easily  remove  those  that  flow 


120  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


as  readily  as  water,  but  many  of  the  reser- 
voirs contain  oil  as  viscous  as  road  tar. 
Miscible  processes  use  injected  chemicals 
that  blend  with  the  crude  oil  to  form  mix- 
tures that  flow  more  readily.  The  chemi- 
cals used  include  alcohols,  carbon  dioxide, 
petroleum  hydrocarbons  such  as  propane 
and  butane-propane  mixtures,  and  petrole- 
um gases.  A fluid  such  as  water  is  generally 
used  to  push  a “slug”  of  these  chemicals 
through  the  reservoir  to  mix  vvdth  the 
crude  oil  and  move  it  to  the  surface. 

• Chemicals  are  also  used  in  alkaline  flood- 
ing, polymer  flooding,  and  combined  sur- 
factant/polymer flooding. 

In  alkaline  flooding,  sodium  hydroxide,  sodi- 
um carbonate,  or  other  alkaline  materials  are 
used  to  enhance  the  flow  of  oil.  Neither  natural 
nor  genetically  engineered  micro-organisms  are 
considered  useful  in  this  process. 

Polymer  flooding  is  a recent  apparently  suc- 
cessful method  of  recovery.  It  depends  on  the 
ability  of  certain  chains  of  long  molecules, 
known  as  polymers,  to  increase  the  viscosity  of 
water.  Instead  of  altering  the  characteristics  of 
the  crude  oil,  the  aim  is  to  make  the  injected 
water  more  capable  of  displacing  it. 

In  the  combined  surfactant/polymer  flooding 
technique,  a detergent-like  material  (surfactant) 
is  used  to  loosen  the  oil  from  its  surrounding 
rock,  while  water  that  contains  a polymer  to  in- 
crease its  viscosity  is  used  to  drive  the  oil  from 
the  reservoir.  (See  figure  27.) 

• Other  EOR  methods  include  many  novel 
possibilities,  such  as  the  injection  of  live 
micro-organisms  into  a reservoir.  These 
may  produce  any  of  the  chemicals  used  in 
miscible  and  chemical  processes,  from  sur- 
factants and  polymers  to  carbon  dioxide. 
One  target  for  EOR  is  the  half  million  strip- 
per wells  (producing  less  than  10  barrels 
per  day  (bbl/d)  in  the  United  States. 

MICROBIAL  PRODUCTION  OF  CHEMICALS 
USED  IN  EOR 

EOR  methods  that  use  chemicals  tend  to  be 
expensive  because  of  the  cost  of  the  chemicals. 
Nevertheless,  potentially  useful  polymers  were 


Figure  27. — Chemical  Flooding  Process 


Injection  fluids  Oil  and  water 


□ Drive  water  zone  CD  Surfactant  slug  zone 
IZD  Water/polymer  IH  Oil  and  water  zone 
zone 

SOURCE'  Office  of  Technology  Assessment,  Enhanced  Oil  Recovery  Potential 
i(i  the  United  States  (Washington,  D C.:  U S Government  Printing  Of- 
fice, January  1978). 

found  in  the  early  1960’s  and  ha\’e  sinct*  heiMi 
responsible  for  the  recovery  of  mort*  than  2 
million  bhl.  Polymers  such  as  polyacrylamide 
and  xanthan  gum  can  increase  th(>  \ iscosity  ol 
water  in  concentrations  as  low  as  one  part  in  a 
thousand.  Xanthan  gum  is  readily  made  in  large 
quantities  by  micro-organisms.  Different  straiuh 
of  Enterobacter  aerogenes  product'  a w ide  \ arie- 
ty  of  other  polymers.  A useful  hiopolymei’— one 
formed  by  a biological  process— might  he  de- 
signed specifically  to  improvt*  oil  recov cry. 

Xanthan  gum,  product'd  by  Xanlhomonas 
campestris  and  currently  marketf'd  by  the  kelco 
division  of  Merck  &.  Uo.,  Inc.,  is  useful  hut  far 
from  ideal  for  oil  recovery.  While  it  has  ex- 
cellent viscous  properties,  it  is  also  very  expen- 
sive. Furthermore,  unless  it  is  exceptionally 
pure,  it  can  plug  reservoir  pon*s,  since  the  fluid 
often  has  to  travel  through  hundicds  of  meters 
of  fine  pores.  To  avoid  such  plugging,  the  fluid 
must  he  filtered  to  remove  bacterial  dehi  is  be- 
fore it  is  injected. 

Nevertheless,  micro-organisms  can  he  -.e 
lected  or  genetically  (‘iigineered  to  overcome 
many  obvious  difficulties.*  With  im|)roved 
properties,  polysaccharides  (polymeric  -aigar-.! 

'A  good  organi.sm,  loi'  (■\ani|)li‘  miglii  li.nr  iln'  i.  ^ „ 

desired  properties:  ooo|)athogeoir  to  horn, ms  pi, mis  i,  ■■■  ...il 
rapiti  growlh  on  simple,  cheap  i .iw  m.ilei'i.ds  e.i-.e  of  ..  p C:  n 
from  its  |)rodii(Ts;  limited  detriment.d  ellei  I on  le.r:  .:,  - 
plugging:  easy  disposal  ofcells  eg  h\ pnidoi  I i 1 1 lit-  ,ilii  n-,  ■ 


Ch.  7— The  Use  of  Genetically  Engineered  Micro-Organisms  in  the  Environment  • 121 


obtained  bv  microbial  t'ermentation  could  com- 
pete with  those  obtained  from  alternative 
sources,  especially  seau  eed.  C'ontrolled  fermen- 
tation is  not  affected  by  mai'ine  pollution  and 
weather,  and  pi'oduction  could  be  geared  to 
market  demand. 

Biological  processes  have  disadvantages  pri- 
marily in  the  costs  of  appro[)riate  raw  materials 
and  in  the  need  foi-  large  (juantities  of  solvent. 
(Current  efforts  to  find  cbeapei’  raw  materials, 
such  as  sugar  beet  pulp  and  starch,  show  prom- 
ise. The  need  for  solv  ents  to  precipitate  and  con- 
centrate the  polymers  before  shipment  from 
plant  to  field  can  be  circumvented  by  producing 
them  onsite. 

I Micro-organisms  can  also  produce  substances 

j like  butyl  and  propyl  alcohols  that  can  be  used 

I as  cosurfactants  in  PX)K.  It  has  been  calculated 
that  if  n-butanol  were  used  to  produce  crude  oil 
at  a level  of  5 percent  of  l^S.  consumption,  2 
billion  to  4 billion  lb  per  year— or  four  to  eight 
times  the  current  butanol  production— would 
be  required.  Micro-organisms  capable  of  pro- 
ducing such  surfactants  have  been  identified, 
and  genetically  superior  strains  were  isolated 
several  decades  ago  at  the  Northern  Regional 
Research  Udioratories  in  Illinois.  Other  chem- 
icals, such  as  alcohols  that  increase  the  rate  of 
formation  and  stability  of  chemical/crude  oil 
mi.xtures  and  the  agents  that  help  prevent  pre- 
cipitation of  the  surfactants,  have  also  been  pro- 
duced by  microbial  systems. 

The  uncertainties  of  the  technical  and  eco- 
nomic parameters  are  compounded  by  the  lack 
of  sufficient  field  experiments.  Laboratory  tests 
cannot  be  equated  with  conditions  in  actual  oil 
wells.  Each  oil  field  has  its  own  set  of  character- 
istics— salinity,  pH  (acidity  and  alkalinity), 
temperature,  porosity  of  the  rock,  and  of  the 
crude  oil  itself— and  an  injected  chemical  be- 
haves differently  in  each  setting.  In  most  cases, 
not  enough  is  known  about  a well’s  characteris- 
tics to  predict  the  nature  of  the  chemical/crude 
oil  interaction  and  to  forecast  the  efficiency  of 
oil  recovery. 


use  water  available  at  site:  grou-th  under  conditions  that  discour- 
age the  growth  of  unwanted  micro-organisms:  no  major  problems 
in  culturing  the  bacterium:  and  genetic  stability. 


I.\  SlTl'  USE  OF  MICRO-ORGANISMS 

One  alternative  to  growing  micro-organisms 
in  large  fermenters  then  extracting  their  chem- 
ical products  and  injecting  them  into  wells,  is  to 
inject  the  micro-organisms  directly  into  the 
wells.  They  could  then  produce  their  chemicals 
in  situ. 

I’nfortunately,  the  geophysical  and  geochem- 
ical conditions  in  a reservoir  seldom  favor  the 
growth  of  micro-organisms.  High  temperature, 
the  presence  of  sulfur  and  salt,  low  oxygen  and 
water,  extremes  of  pH,  and  significant  engi- 
neering hurdles  make  it  difficult  to  ov'ercome 
these  limitations.  The  micro-organisms  must  be 
fed  and  the  microenvironment  must  be  care- 
fully adjusted  to  their  needs  at  distances  of  hun- 
dreds to  thousands  of  feet.  The  oil  industry  has 
already  had  discouraging  experiences  with 
micro-organisms  in  the  past.  In  the  late  1940’s, 
for  instance,  the  injection  of  sulfite-reducing 
micro-organisms,  along  with  an  inadvertently 
high-iron  molasses  as  a carbon  source,  resulted 
in  the  formation  of  iron  sulfide,  which  clogged 
the  rock  pores.  One  oil  company  developed  a 
yeast  to  break  down  petroleum,  but  the  size  of 
the  yeast  cells  (5  to  10  micrometers,  /im)  was 
enough  to  clog  the  l-/xm  pores. 

Nevertheless,  information  from  geomicrobi- 
ologv'  suggests  that  this  approach  is  w'orth  pur- 
suing. Preliminary  field  tests  have  also  been  en- 
couraging. The  injection  of  1 to  10  gal  of  Bacillus 
or  Clostridium  species,  along  with  a water- 
suspended  mixture  of  fermentable  raw  materi- 
als such  as  cattle  feed  molasses  and  mineral 
nutrients,  has  resulted  in  copious  amounts  of 
carbon  dioxide,  methane,  and  some  nitrogen  in 
reservoirs.  The  carbon  dioxide  made  the  crude 
less  viscous,  and  the  other  gases  helped  to 
repressurize  the  reservoir.  In  addition,  large 
amounts  of  organic  acids  formed  additional  car- 
bon dioxide  through  reactions  with  carbonate 
minerals.  The  production  of  microbial  sur- 
factants further  aided  the  process. 

Although  previous  assessments  have  argued 
that  reservoir  pressure  is  a significant  hin- 
drance to  the  growth  of  micro-organisms,  more 
recent  studies  indicate  the  contrary.  The  micro- 
organisms must,  however,  be  selected  for  in- 
creased salt  and  pH  tolerance. 


122  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


EOR  AND  GENETIC  ENGINEERING 

The  current  research  approach,  funded  by 
the  Department  of  Energy  (DOE)  and,  independ- 
ently, by  various  oil  companies,  is  a two-phase 
process.  The  first  phase  is  to  find  a micro- 
organism that  can  function  in  an  oil  reservoir 
environment  with  as  many  of  the  necessary 
characteristics  as  possible.  The  second  is  to  alter 
it  genetically  to  enhance  its  overall  capability. 

The  genetic  alteration  of  micro-organisms  to 
produce  chemicals  used  in  EOR  has  been  more 
successful  than  the  alteration  of  those  that  may 
be  used  in  situ.*  However,  recombinant  DNA 
(rDNA)  technology  has  not  been  applied  in  ei- 
ther category.  All  efforts  have  employed  artifi- 
cially induced  or  naturally  occurring  mutations. 

CONSTRAINTS  TO  APPLYING  GENETIC 
ENGINEERING  TECHNOLOGIES  IN  EOR 

The  genetic  data  base  for  micro-organisms 
that  produce  useful  polysaccharides  is  weak. 
Few  genetic  studies  have  been  done.  Hence,  the- 
oretically plausible  approaches  such  as  transfer- 
ring enzyme-coding  plasmids  (see  ch.  2)  for 
polysaccharide  synthesis,  cannot  be  seriously 
contemplated  at  present.  Only  the  crudest 
methods  of  genetic  selection  for  desirable  prop- 
erties have  been  used  thus  far.  They  remain  the 
only  avenue  for  improvement  until  more  is 
learned  about  the  micro-organism’s  genetic 
mechanisms. 

The  biochemical  data  base  for  the  character- 
istics of  both  the  micro-organisms  and  their 
products  is  also  lacking.  The  wide  potential  for 
chemical  reactions  carried  out  by  microbes  re- 
mains to  be  explored.  At  the  same  time,  a sys- 
tem must  be  devised  to  allow  easy  characteriza- 
tion, classification,  and  comparison  of  products 
derived  from  a variety  of  micro-organisms. 

The  physical  data  base  for  oil  reservoirs  is 
limited.  The  uniqueness  of  each  reservoir  sug- 
gests that  no  universal  micro-organism  or  meth- 
od of  oil  recovery  will  be  found.  Compounding 

’Some  of  the  goals  have  been  to:  improve  polymer  properties  to 
enhance  their  commercial  applicability;  improve  polymer  produc- 
tion (a  major  mistake  has  been  to  reject  a micro-organism  in  the 
initial  screening  because  its  level  of  production  was  too  low);  im- 
prove culture  characteristics,  e.g.,  resistance  to  phage,  rapid 
growth,  ability  to  use  cheaper  raw  materials;  and  eliminate  en- 
zymes that  naturally  degrade  the  polymers. 


this  problem  is  the  lack  of  sufficient  physical, 
chemical,  and  biological  information  about  the 
reservoirs,  without  which  it  is  difficult  to  see 
how  a rational  genetic  scheme  can  be  con- 
structed for  strains.  Clearly,  the  activities  of 
micro-organisms  under  specified  field  condi- 
tions cannot  be  studied  unless  researchers 
know  what  the  appropriate  conditions  are. 

Three  institutional  obstacles  exist.  First,  publi- 
cation in  this  field  is  limited  because  most  re- 
search is  carried  out  in  the  commercial  world 
and  remains  largely  confidential.  Second,  nei- 
ther the  private  nor  the  public  sector  has  been 
enthusiastic  about  the  potential  role  of  micro- 
organisms in  EOR.  The  biological  apjtroach  has 
only  recently  been  given  consideration  as  a way 
to  advance  the  state  of  the  art  of  the  technology, 
and  most  oil  companies  still  ha\  e limited  staffs 
in  microbiology.  To  date,  DOE’s  Division  of 
Fossil  Fuel  Extraction  has  conducted  the  main 
Federal  effort.  Third,  any  effoi't  to  use  micro- 
organisms must  he  multidisciplinary  in  nature. 
Geologists,  microbiologists  (incliuling  mici'ohial 
physiologists  and  geneticists),  chemists,  and 
engineers  must  interact  to  evoke  successful 
schemes  of  oil  recovei'v.  'Fhus  far,  such  t(>ams 
do  not  exist. 

Environmental  and  legal  concerns  have  also  in- 
hibited progress.  Microbial  EOK  methods  usual- 
ly require  significant  (juantities  of  fresh  wat(>r 
and  thus  may  compete  with  municipal  and  agri- 
cultural uses.  Furthermore,  the  use  of  micro- 
organisms introduces  concerns  for  safety.  .All 
strains  of  Xanthomonas,  which  produce  .xanthan 
gum  polymer,  are  plant  pathogens  Other 
micro-organisms  with  potential,  such  as  Scleroti- 
um  rolfii  and  various  species  of  Aureobasidium 
have  been  associated  with  lung  disease  and 
wound  infections,  respectively. 

Immediate  environmental  and  legal  concerns, 
therefore,  arise  from  the  |)otential  risks  .issoci- 
ated  with  the  release  of  micro-organisms  into 
the  environment.  When  th(*v  naturallv  c.uisi* 
disease  or  environmental  disru|)tion.  tln’ir  use  is 
clearly  limited.  And  wIkmi  they  do  not  genetic 
engineering  raises  the  possibility  that  they 
might.  Sucli  concerns  have  ri*duced  the  jiriv.ite 
sector’s  enthusiasm  for  attempting  genetic 


Ch.  7 — The  Use  of  Genetically  Engineered  Micro-Organisms  in  the  Environment  • 123 


engineering.  (See  ch.  10  for  a moi’e  detailed 
discussion  of  risk.) 

OE.NETIC  E.\(;i\EEKI.\(;  OF  MICKO-OKO.AMSMS 
FOK  I'SE  I,\  OrilEK  ASPECTS  OF  OIL 
RECOVER V A.M)  TREATMENT 

I'wo  other  aspects  of  microhial  {)hvsiolog\’ 
deserve  attention:  the  microhial  production  of 
oil  muds  or  di'ill  luhricants,  and  the  treatment 
of  oil  once  it  has  been  recovered.  Drilling  muds 
are  suspensions  of  clays  and  other  materials 
that  serve  both  to  lubricate  the  drill  and  to 
counterbalance  the  upu'aixl  pressui’e  of  oil.  Mi- 
ci'ohially  pi'oduced  polysaccharides  have  been 
dev  eloped  for  this  use.  K.x.xon  holtls  a patent  on 
a formulation  based  on  the  production  of  xan- 
than  gum,  from  Xanthomonas  campestris,  while 
the  Pillshui'v  Co.  has  developed  a [)olysac- 
charide  (glucan)  from  various  s[)ecies  of  Scler- 
otium.  .At  least  two  of  the  small  genetic 
engineering  firms  have  begun  I'esearch  pro- 
grams to  develop  biologically  pi'oduced  polysac- 
charides with  the  desired  lubricant  qualities. 

Interest  in  the  postrecovei’v  mici'obial  treat- 
ment of  oil  after  its  extraction  centers  around 
the  ability  of  micro-organisms  to  remove  un- 


tlesirahle  contituents  from  the  crude  oil  itself. 
As  an  indication  of  recent  progress,  three  dis- 
tinct microhial  systems  have  been  developed  to 
help  remove  aromatic  sulfur-containing  mate- 
rial, a major  impurity. 

Oi'ervieiv  of  genetic  engineering  in 
mining  and  oil  recovery 

The  underlying  technical  problem  with  the 
use  of  genetically  engineered  organisms  in 
either  mining  or  oil  recovery  is  the  magnitude 
ol  the  effort,  in  both  cases,  large  areas  of  land 
and  large  volumes  of  materials  (chemicals,  flu- 
ids, micro-organisms)  must  he  used.  The  results 
ol  testing  any  new  micro-organism  in  a labora- 
tory cannot  automatically  he  extrapolated  to 
large-scale  applications.  The  change  in 
magnitude  is  fui'ther  complicated  by  the  lack  of 
rigid  controls.  Linlike  a large  fermenter  whose 
temperature,  pfl,  and  other  characteristics  can 
he  carefully  regulated,  the  natural  environment 
cannot  he  controlled.  Nevertheless,  despite  the 
formidable  obstacles,  the  potential  value  of  the 
products  in  these  areas  assures  continuing  ef- 
forts. 


Pollution  control 


Life  is  a cycle  of  synthesis  and  degradation- 
synthesis  of  complex  molecules  from  atoms  and 
simple  molecules  and  degradation  by  bacteria 
yeast,  and  fungi,  back  to  simpler  molecules  and 
atoms  when  organisms  die.  The  degradation  of 
complex  molecules  is  an  essential  part  of  life. 
U'ithout  it,  “.  . . w e’d  be  knee-deep  in  dino- 
saurs.”* A more  quantitative  statement  is  equal- 
ly thought  provoking.  Livestock  in  the  United 
States  produce  1.7  billion  tons  of  manure  an- 
nually. Almost  all  of  it  is  degraded  by  soil  micro- 
organisms. 

For  a long  time  people  have  exploited  micro- 
bial life  forms  to  degrade  and  detoxify  human 
sewage.  Now,  on  a smaller  scale,  science  is 


'R.  B.  Grubbs,  "Bacterial  Supplementation,  What  It  Can  and  Can- 
not Do."  oral  presentation  to  the  Ninth  Engineering  Foundation  on 
Environmental  Engineering  in  the  Food  Processing  Industry,  1979 
(Available  from  Flow  Laboratories,  Inc.,  Rock\  ille,  Md.l 


beginning  to  use  micro-organisms  to  deal  with 
the  pollution  problems  presented  by  industrial 
toxic  wastes.  Chemicals  in  their  place  can  be 
useful  and  beneficial;  out  of  place,  they  can  be 
polluting. 

Pollution  problems  can  be  divided  into  two 
categories;  those  that  have  been  present  for  a 
long  time  in  the  biosphere— e.g.,  most  hydro- 
carbons encountered  in  the  petroleum  industry 
and  human  and  animal  wastes— and  those  that 
owe  their  origin  to  human  inventiveness— e.g., 
certain  pesticides.  Chemicals  of  both  sorts, 
through  mishap,  poor  planning,  or  lack  of 
knowledge  at  the  time  of  their  application 
sometimes  appear  in  places  where  they  are 
potentially  or  actually  hazardous  to  human 
health  or  the  environment. 

Pollution  can  he  controlled  hy  microbes  in 
two  ways;  hy  enhancing  the  growth  and  activity 


124  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


of  microbes  already  present  at  or  near  the  site 
of  the  pollution  problem^  and  by  adding  more 
(sometimes  new)  microbes  to  the  pollution  site. 
The  first  approach  does  not  provide  an  oppor- 
tunity for  applying  genetics^  but  an  example  will 
indicate  how  it  functions. 

Enhancing  existing  microbial 
degradation  activity 

Sun  Oil  successfully  exploited  indigenous 
microbes  to  clean  up  a 6,000  gal  underground 
gasoline  spill  that  threatened  the  water  supply 
of  a town  in  Pennsylvania. ^ ^ First,  engineers 
drilled  wells  to  the  top  of  the  water  table  and 
used  pumps  to  skim  gasoline  from  the  water 
surface.  About  half  the  gasoline  was  removed  in 
this  fashion,  but  company  calculations  showed 
that  dissipating  the  remaining  gasoline  would 
require  about  100  years.  To  speedup  the  proc- 
ess, it  was  decided  to  encourage  the  growth  of 
indigenous  bacteria  that  could  degrade  the 
gasoline. 

Pollution-control  microbes,  like  all  organisms, 
require  a number  of  different  elements  and 
compounds  for  growth.  If  the  amount  of  any 
nutrient  is  limited,  the  microbe  will  not  be  able 
to  metabolize  the  pollutant  at  the  fastest  rate. 
The  cleanup  depended  on  increasing  the 
growth  rate  of  the  bacteria  by  supplying  them 
with  additional  nutrients.  In  the  case  of  the 
gasoline-degrading  bacteria,  the  gasoline  al- 
ready supplied  the  hydrocarbon,  but  the  water- 
gasoline  environment  was  deficient  in  nitrogen, 
phosphate,  and  oxygen.  Those  three  nutrients 
were  pumped  down  to  the  water  table,  bacterial 
growth  increased,  and  the  gasoline  was  metabo- 
lized into  innocuous  chemicals  by  the  bacteria. 
As  a result,  it  was  degraded  in  a single  year. 

Adding  microbes  to  clean  up  pollution 

Genetics  may  have  important  applications  in 
approaches  to  pollution  control  that  depend  on 

^R.  L.  Raymond,  V.  W.  Jamison,  J.  O.  Hudson,  "Beneficial  Stimu- 
lation of  Bacterial  Activity  in  Groundwaters  Containing  Petroleum 
Products,"  AIChE  symposium  series  73:390-404,  1976. 

^V.  W.  Jamison,  R.  L.  Raymond,  J.  O.  Hudson,  "Biodegradation  of 
High-Octane  Gasoline,"  Proceedings  of  the  Third  International  Bin- 
degradation  Symposium,  J.  M.  Sharpley  and  A,  M.  Kaplan  (eds.) 
(City????  : Applied  Science  Publishers,  1976). 


adding  microbes  to  the  pollution  site.  Three 
firms— Flow  Laboratories,  Polybac  Corp.,  and 
Sybron/Biochemicals  Corp.— sell  microbes  for 
such  use.  Two  companies  select  bacteria  for  en- 
hanced degradation  acti\itv  and  two  mutate 
bacteria  to  the  same  end,  hut  none  of  the  three 
firms  currently  uses  genetic  engineering  tech- 
niques. 

Some  "formulations”  (mixtures)  of  bacteria 
are  designed  to  degrade  particular  pollutants, 
such  as  one  that  was  used  to  digest  the  HOO.OOO 
gal  of  oily  water  that  lay  in  the  bilges  of  the* 
Queen  Mary.  After  a 6-week  treatment  with  the 
formulation,  the  water  from  the  bilges  was 
judged  safe  for  disposal  into  the  Long  Beach, 
Calif.,  harbor.  It  was  discharged  without  caus- 
ing an  oil  slick  or  harming  mai’ine  life.-*  F1o\n 
Laboratories  markets  its  ser\  ic(‘s  to  companies 
with  industrial  pollution  pi'ohlems.  It  iincsti- 
gates  the  problem,  develops  a formulation  to 
degrade  the  pollutants,  and  sells  it. 

In  addition  to  industrial  pollution  pi'ohlems. 
Flow'  markets  its  products  and  ser\  i('(\s  for  u.se 
in  sewerage  systems,  which  collect  and  hold 
human  wastes  to  facilitat(>  degradation  and  de- 
toxification. Sludge  hact(M’ia  in  sewerage  plants 
degrade  the  w^aste,  hut  they  are  not  pre.siMit  in 
the  lines  that  cany  wastes  to  the  treatiiuMit 
plant.  As  a result,  gr(\ises  and  oils  from  fat  dis- 
carded through  gai'hage  dis|)osals  and  I rom  cos- 
metic oils  and  creams  coat  tlie  inside  of  sewer- 
age lines  and  reduc(^  their  carrying  ca|)aeity.’ 

Cities  have  resisted  using  added  microbes  in 
sewerage  systems.  Standard  te.\tl)ooks  simply 
state  that  the  ideal  hactcM'ia  w ill  establish  them- 
selves in  a w'ell-|)lann('d  and  well-managed  sys- 
tem. The  idea  that  ''better"  bacteria  can  l)c 
added  to  imjjrovfj  th(‘  plant  operation  is  not 
readily  accepted. 

Fhe  value  of  adding  bacteria  to  large  sewer- 
age sytems  has  not  been  ade(|uatel\  tested 
Because  of  the  size  of  municipal  .systems  (w  Inch 
already  contain  tons  of  sludge  bacterial,  some 
have  argued  that  adding  a tew  .iddilion.il 


*.\uun.  T.nvironmental  Scirntf  anil  t rrhnolog\  1.11  isn  C ' 

•’R  I-..  Kirkup  iind  I.  R Srlsmi.  ( il\  I ikIiW  (.n-.i  -.  .ind  0,1. 
Pi'ohlem.s  in  Scwim'  SysU-ms.  Ptihlii  Works  Magarmr  (Ki.'l-. 
1977. 


Ch.  7 — The  Use  of  Genetically  Engineered  Micro-Organisms  in  the  Environment  • 125 


pounils  of  bac  teria  is  unlikely  to  ha\  e any  effect, 
rhus  far.  the  Km  ironmental  Pi  otection  .Agency 
(KI’.A)  lias  not  reconimendetl  adding  bacteria  to 
municipal  sNstems:  bo\ve\ei‘,  KI’.A  suggests  that 
tbe\  might  be  useful  in  smaller  installations  and 
foi’  specific  problems  in  lai'ge  systems. 

l)iy  foi'mulations  are  available  for  use  in 
cleaning  drains  and  pipes  in  smaller  installa- 
tions. such  as  restaurants  and  other  food  proc- 
essing facilities.  In  restaui'ants.  the  bacteria  are 
added  to  the  drain  at  the  end  of  the  workday. 
Bacteria  have  been  selected  foi-  their  inability  to 
produc'e  bydi’ogen  sulfide,  which  means  that 
the  degrading  process  does  not  produce  the  un- 
pleasant odors  fiecjuently  encountered  in  the 
digestion  of  oils  and  fats.® 

As  of  N'ovembei'  1979,  the  pollution  control 
industrv  had  few  plans  for  the  genetic  manipu- 
lation of  bacteria,  e.xcept  for  the  selection  of 
naturally  occurring  better  [)erformers.  Clon- 
sumer  resistance  to  mutants”  is  a factor  that 
discourages  the  move  to  microbial  genetics. 
Probably  even  more  important  is  the  high  cost 
of  establishing  and  maintaining  microbial  genet- 
ics lalmratories.  It  has  been  estimated  that  the 
cost  of  carrying  a single  Ph.  D.  microbial  geneti- 
cist is  over  $100,000  annually.'  This  e.xpense  is 
quite  high  relative  to  the  $2  million  to  $4  million 
sales  of  all  biological  pollution  control  com- 
panies in  1978.® 

Resistance  to  the  use  of  genetically  manip- 
ulated bacteria  is  not  universal.  Many  industrial 
wastes  are  o.xidized  to  nontoxic  chemicals  by 
biological  treatment  in  aerated  lagoons.  The 
process  depends  on  the  presence  of  microbes  in 
the  lagoons:  over  time,  those  that  grow  best  on 
the  wastes  come  to  dominate  the  microbial  pop- 
ulations. Three  companies  now  sell  bacteria 
that  they  claim  outperform  the  indigenous 
strains  found  in  the  lagoons.  E.g.,  the  Polybac 


‘.Anon..  "Clean  That  Sewage  System  VV  ith  Bugsl " Environmental 
. Science  and  Technology  13:1198-1199.  1979 

'.Anon..  " Biotechnology  DN.A  Research  E.\penditures  in  L'.S.  May 
i Reach  S500  .Million  in  1980.  W ith  About  S 150-200  Million  for  Coni- 
I mercial  Products.  " Hill  told.  Drug  Research  Reports,  " The  Blue 
i Sheet  Vlay  28  1980  p.  22. 

‘Anon..  Business  Week.  July  5.  1976.  p.  280:  Chemical  Week 
I 121:47,  1977:  and  Food  Engineering  49:138.  1977,  cited  in  T.  Gass- 
>ner.  "Microorganisms  for  Waste  Treatment,  " Microbial  Technol- 
: ogy,  2 ed..  vol.  II.  (London:  .Academic  Press,  1979),  pp.  211-222. 


Corp.  has  sold  its  products  to  all  seven  Exxon 
biological  waste  treatment  plants  to  treat  chem- 
ical wastes.  One  of  its  formulations  has  been 
used  to  degrade  toxic  dioxins  from  an  herbicide 
spill.  One  month’s  treatment  with  the  bacterial 
formulation  reduced  the  orthochlorophenol 
concentration  from  600  to  25  ppm  in  a 20,000- 
gal  lagoon.® 

Syhroiv'Biochemical,  a division  of  Sybron 
C!orp.,  sells  cultures  of  bacteria  that  are  in- 
tended to  aid  in  the  biological  oxidation  of  in- 
dusti'ial  wastewater;  this  company  also  lists  20 
different  cultures  for  application  to  specific 
wastes.  Patent  number  4,199,444  was  granted 
on  .April  22,  1980,  for  a process  involving  the 
use  of  a mutant  bacterial  culture  to  decolor 
waste  water  produced  in  Kraft  paper  process- 
ing."’ Other  patents  are  pending  on  a mixture  of 
two  strains  that  degrade  grease  and  a strain  that 
degrades  "nonhiodegradable”  detergents.” 

There  is  disagreement  about  the  value  of  add- 
ing microbes  to  decontaminate  soils  or  waters. 
One  point  of  view  argues  that  serious  spills  fre- 
quently sterilize  soils,  and  that  adding  microbes 
is  necessary  for  any  biodegradation.  The  other 
contends  that  encouraging  indigenous  microbes 
is  more  likely  to  succeed  because  they  are  ac- 
climated to  the  spill  environment.  Added  bac- 
teria have  a difficult  time  competing  with  the 
already-present  microbial  flora.  In  the  case  of 
marine  spills,  bacteria,  yeast,  and  fungi  already 
present  in  the  water  participate  in  degradation, 
no  one  has  been  able  to  demonstrate  the  useful- 
ness of  added  microbes. 

Commercial  applications — market  size 
and  prospects 

The  estimated  market  size  of  pollution-con- 
trol biological  products  in  1978  was  $2  million  to 
S4  million,  divided  among  some  20  companies, 

‘See  footnote  6. 

“L.  Davis.  J.  E.  Blair,  and  C.  VV.  Randall,  "Communication: 
Development  of  Color  Removal  Potential  in  Organisms  Treating 
Pulp  and  Paper  VV'astevvater,"  J.  Water  Pollution  Control  Fed.,  Feb- 
ruary 1978,  pp.  382-385. 

"P.  Spraher  and  N'.  Tekeocgak,  "'Foam  Control  and  Degradation 
of  Nonionic  Detergent,"  Industrial  Wastes,  January/February  1980: 
L,  David,  J.  E.  Blair,  and  C.  Randall,  "Mixed  Bacterial  Cultures  Leak 
'Non-Biodegradable'  Detergent,"  Industrial  Wastes,  May/June 
1979. 


126  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


and  the  potential  market  was  estimated  to  be  as 
much  as  $200  milliond^  These  estimates  can  be 
compared  to  Polybac’s  own  sales  records.  In 
1976,  its  first  year,  its  sales  totaled  $0.5  million 
and  in  1977,  $1.0  million.  It  expects  to  reach  $5 
million  in  1981. 

To  date  genetically  engineered  strains  have 
not  been  applied  to  pollution  problems.  At  least 
one  prominent  genetic  engineering  company 
has  decided  not  to  enter  the  pollution  control 
field,  concluding  that  it  was  improbable  that 
added  microbes  could  compete  with  indigenous 
organisms.  More  specifically,  the  possiblity  of 
liability  problems  make  the  approach  even  less 
attractive.  Pollution  control  requires  that  “new” 
life  forms  be  released  into  the  environment, 
which  is  already  seen  as  precariously  balanced. 
Such  new  forms  might  cause  health,  economic, 
or  environmental  problems.  The  problems  of 
liability  that  might  arise  from  such  applications 
are  enough  to  deter  entrepreneurs  from  con- 
templating work  in  the  field  at  this  time. 

An  additional  reason  for  the  reluctance  of 
some  companies  to  engage  in  this  activity  is  that 
the  opportunities  for  making  money  are  limited. 
Selling  microbes,  rather  than  their  products, 
may  well  be  a one-shot  opportunity.  The  mi- 
crobes, once  purchased,  might  be  propagated 
by  the  buyer.  Nevertheless,  at  least  two  small 
companies  have  announced  that  they  are  pursu- 
ing efforts  to  use  genetic  engineering. 

The  low-key  efforts  in  this  field  might  accel- 
erate quickly  if  a significant  breakthrough  oc- 
curred. To  date,  no  “new”  organism  has  ap- 
peared that  will  degrade  previously  intractable 
chemicals.  The  effect  of  such  a development 
might  be  enormous. 


Genetic  research  in  pollution  control 

The  Oil  and  Hazardous  Materials  Spills 
Branch  of  EPA  currently  supports  research 
aimed  at  isolating  organisms  to  degrade  three 
specific  chemical  compounds.  The  work  is  being 
carried  out  on  contract;  as  of  November  1979, 
no  field  trials  of  the  organisms  had  been  under- 


'^See footnote  8. 


taken.  Two  of  the  toxic  chemicals,  pentachoro- 
phenol  and  hexachlorocyclopentadiene,  are 
relatively  long-lived  compounds  and  present 
long-term  problems.  A fungus  and  a bacterium 
that  can  degrade  the  first  compound  ha\  e been 
isolated,^®  and  Sybron/Biochemical  already  sells 
a culture  specifically  for  pentachlorophenol 
degradation.  The  third  toxic  compound  is  meth- 
yl parathion.  Its  inclusion  is  more  difficult  to 
understand,  since  it  is  degraded  within  a few 
days  after  its  application  as  a pesticide. 

Efforts  have  been  made  to  isolate  bacteria 
that  can  degrade  (2,4-dichlorophenoxy)  acetic 
acid  (2,4-D)  and  (2,4,5-trichlorophenoxy)  acetic 
acid  (2,4, 5-T),  the  components  of  Agent 
Orange. Strains  of  the  bacterium  Alcaligenes 
paradoxus  rapidly  degrade  2,4-U,  and  the 
genetic  information  for  the  degradation  acti\  ity 
has  been  located  on  a plasmid.  The  inv  estigator 
who  found  that  strain,  while  optimistic  about 
the  opportunities  for  isolating  and  transferring 
other  resistance  genes,  has  been  unable  to  find 
a bacterium  that  degrades  2, 4, 5-1'  oi'  its  very 
toxic  contaminant,  2,3,7,8-tetrachlorodihenzo- 
para-dioxin  (TCDD  or  dioxin). 

By  far  the  best  known  research  in  this  area  is 
that  of  Dr.  Ananda  M.  Chaki  ahai  ty  who  iMigi- 
neered  two  strains  of  Pseudonionas,  each  of 
which  has  the  ability  to  degrade  th('  four  class(\s 
of  chemicals  found  in  oil  spills,  (diakrahartv 
began  with  four  different  strains  of  Pseudo- 
monas. None  of  them  presented  a threat  to 
human  health,  and  each  could  d(‘gi  ade  one  of 
the  four  classes  of  chemicals.  His  research 
showed  that  the  genes  controlling  the  degi  ading 
activities  were  located  on  plasmids,  faking  ad- 
vantage of  the  relative  ease  of  moving  such 
genes  among  bacteria,  he  produced  two  recom- 
binant bacteria. 

Chakraharty  has  presented  ev  idence  that  his 
bacterium  degrades  complex  petioleum  mix- 
tures such  as  crude  oil  or  Bunkj'i’  (' " oil.  and  In* 


'^N.  K.  rhuma.  P.  K.  O'Nrill,  S (.  Ili-msnliT  .met  H ^ V.ilrnlmf 
"Laboratory  Feasil)ilily  and  Pilot  Plant  Stndii-s  Novel  ItiodeKi  .id.i 
tion  Processes  for  the  Ultimate  Dispos.il  ol  Spilli-d  li.i/.n dme. 
Materials,"  National  I-'.nv  ironmi'nt  Keseai  c h I enti-i  t s I muon 
mental  Protection  Af'eney,  Cincinnati,  Ohio,  I!I7H 

‘“J.  M.  Pemberton,  "Pesticide  De^radin^  PlaMtiuK  V lliolof;M  .d 
Answer  to  Knvironmental  Pollution  by  Phenow  berim  id<  ' tei/.,,, 

8:202-20.';,  1979. 


Ch.  7 — The  Use  of  Genetically  Engineered  Micro-Organisms  in  the  Environment  • 127 


has  proposed  a method  tor  using  it  to  clean  up 
oil  s[)ills.  The  bacteria  are  to  he  grown  in  the 
lahoratorv,  mixed  with  sti'aw,  and  dried.  The 
hacteria-coated  straw  can  he  stored  until 
needed,  then  dropped  from  a ship  or  airci'aft 
onto  oil  spills.  The  straw  ahsoi'hs  the  oil  and  the 
bacteria  degrades  it.'*  To  completely  cleanu[)  a 
spill  w ill  prohahh’  retjuire  mechanical  efforts  in 
addition  to  the  biological  attack.  It  was  the  pro- 
duction of  one  of  (diakraharty’s  strains  that  led 
to  the  Supreme  (!ourt  decision  on  "the  patenting 
of  life.”  (See  ch.  12  for  further  details.) 

The  essential  difference  between  the  well- 
publicized  Chakraharty  approach  and  a less 
well-known  one  is  that  all  the  desired  acti\  ities 
in  C'hakrabarty’s  approach  are  combined  in  a 
single  organism;  while  in  the  other  method, 
bacteria  hearing  single  activities  are  mixed 
together  to  yield  a desired  “formulation.”  In  yet 
another  approach,  Sybron/Biochemical  uses 
mutation  and  selection  to  produce  specialized 
degradation  activities.  It  also  sells  mixed 
cultures  for  some  applications. 

The  single-organism,  multiple-enzyme  system 
has  the  adv  antage  that  ev  ery  bacterium  can  at- 
tack a number  of  compounds.  The  mixed  for- 
mulations allow  the  preferential  proliferation  of 
bacteria  that  feed  on  the  most  abundant  chem- 
ical; then,  as  that  chemical  is  exhausted,  other 
bacteria,  which  nourish  on  the  next  most  abun- 
dant cbemical,  become  dominant.  The  pref- 
erential surv  ival  of  only  one  or  a few  strains  in  a 
mixed  formulation  might  result  in  no  bacteria 
being  available  to  degrade  some  compounds. 
The  multienzyme  bacteria,  on  the  other  hand, 
can  degrade  one  chemical  after  another,  or 
alternativ  ely,  more  than  one  at  the  same  time. 

Federal  research  support  for 
engineering  microbes  to  detoxify 
hazardous  substances 

EPA  currently  limits  its  support  to  research 
aimed  at  selecting  indigenous  microbes,  an  area 


'^Patent  Specification  1 436  573,  May  19,  1976,  Patent  Office, 
London.  England. 


that  has  already  attracted  some  commercial 
research  supjioi't.  Commercial  firms  are  looking 
for  lai'ge-scale  markets,  such  as  sewerage  sys- 
tems, or  commonly  occurring  smaller  markets, 
such  as  gasoline  spills  and  common  industrial 
wastes. 

Whatever  potential  exists  in  identifying, 
growing,  and  using  naturally  occurring  mi- 
crobes tor  pollution  control  pales  beside  the  op- 
portunities ottered  by  engineering  new  ones. 
I'ntortunately,  the  potential  risks  increase  as 
well.  EP,A  has  taken  a preliminary  step  toward 
assessing  the  risks  by  soliciting  studies  to  deter- 
mine what  environmental  risks  may  exist  from 
accidentally  or  deliberately  released  engineered 
microbes. 


Summary 

While  some  unreported  efforts  may  be 
underway,  genetics  bas  apparently  been  little 
applied  to  pollution  abatement.  Nevertheless, 
the  production  of  "new”  life  forms  that  offer  a 
significant  improvement  in  pollution  control  is  a 
possibility.  The  constraints  are  questions  of 
liability  in  the  event  of  health,  economic,  or  en- 
. v'ironmental  damage;  the  contention  that  added 
organisms  are  not  likely  to  be  a significant  im- 
provement; and  the  assumption  that  selling 
microbes  rather  than  products  or  processes  is 
not  likely  to  be  profitable. 

The  factors  that  have  discouraged  develop- 
ments in  this  area  would  probably  become  less 
deterring  if  convincing  evidence  were  found 
that  microbes  could  remove  or  degrade  an  in- 
tractable pollutant.  In  the  meantime,  the  re- 
search necessary  to  produce  marked  improve- 
ments has  been  inhibited.  Overcoming  this  in- 
hibition may  require  a governmental  commit- 
ment to  support  the  research,  to  buy  the 
microbes,  and  to  provide  for  protection  against 
liability  suits.  Such  a governmental  role  would 
be  in  keeping  with  its  commitment  to  protecting 
health  and  the  environment  from  the  toxic  ef- 
fects of  pollutants. 


128  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


Issue  and  Options — Biotechnology 


ISSUE;  How  can  the  Federal  Govern- 
ment promote  advances  in  bio- 
technology and  genetic  engi- 
neering? 

The  United  States  is  a leader  in  applying  ge- 
netic engineering  and  biotechnology  to  indus- 
try. One  reason  is  the  long-standing  commit- 
ment by  the  Federal  Government  to  the  funding 
of  basic  biological  research;  several  decades  of 
support  for  some  of  the  most  esoteric  basic  re- 
search has  unexpectedly  provided  the  founda- 
tion for  a highly  useful  technology.  A second  is 
the  availability  of  venture  capital,  which  has  al- 
lowed the  formation  of  small  innovative  compa- 
nies that  can  build  on  the  basic  research. 

The  argument  for  Government  promotion  of 
biotechnology  and  genetic  engineering  is  that 
Federal  help  is  needed  in  those  high  priority 
areas  not  being  developed  by  industry. 

The  argument  against  such  assistance  is  that 
industry  will  develop  everything  of  commercial 
value  without  Federal  help. 

A look  at  what  industry  is  now  attempting  in- 
dicates that  sufficient  investment  capital  is 
available  to  pursue  specific  manufacturing  ob- 
jectives, such  as  for  interferon  and  ethanol,  but 
that  some  high-risk  areas  that  might  be  of  in- 
terest to  society,  such  as  pollution  control,  may 
need  promotion  by  the  Government.  Other 
areas,  such  as  continued  basic  biological  re- 
search, might  not  be  profitable  soon  enough  to 
attract  industry’s  investment.  Specialized  educa- 
tion and  training  are  areas  in  which  the  Govern- 
ment has  already  played  a major  role,  although 
industry  has  both  supported  university  training 
and  conducted  its  own  inhouse  training. 

OPTIONS: 

A.  Congress  could  allocate  funds  specifically  for 
genetic  engineering  and  biotechnology  R&cD  in 
the  budget  of  appropriate  agencies,  such  as 
the  National  Science  Foundation  (NSF),  the 
U.S.  Department  of  Agriculture  (USD A),  the 
Department  of  Flealth  and  Human  Services 
(DHHS),  the  Department  of  Energy  (DOE),  the 


Department  of  Commerce  (DOC),  and  the  De- 
partment of  Defense  (DOD). 

Congress  has  a long  history  of  recognizing 
areas  of  R&D  that  need  priority  treatment  in 
the  allocation  of  funds.  Biotechnology  has  not 
been  one  of  these.  Even  though  agencies  like 
NSF  receive  congressional  funding,  its  Alter- 
native Biological  Sources  of  Materials  program 
is  one  of  the  few  applied  programs  that  is  not 
congressionally  mandated.  As  a result,  the  fiscal 
year  1980  budget  saw  a reduction  in  the  alloca- 
tion of  funds,  from  $4.1  million  in  1979  to  $2.9 
million.  A congressionally  mandated  program, 
analogous  to  the  successful  NSF  F^irthcjuake 
Hazard  Mitigation  program,  could  he  written 
into  law.  Other  programs,  such  as  the  com- 
petitive grants  program  at  USD  A (or  the  Office 
of  Basic  Biological  Research  at  DOE),  are  also 
modestly  funded. 

Increasing  the  amount  of  money  in  an  ag(Mi- 
cy’s  biotechnology  program  could  bring  criti- 
cism from  other  jirograms  within  each  agcMicy  if 
their  levels  of  funding  are  not  increased  com- 
mensurately.  The  Competiti\e  Gi’ants  Fi’ogram 
at  USDA  has  similar  problems;  those  who  are 
most  critical  of  it  argue  that  it  should  not  take 
funds  from  traditional  programs.  Ne\'ertheless, 
Congress  could  promote  two  typers  of  [)rograms: 
those  with  long-range  payoffs  (basic;  i'(?s('archl, 
and  those  which  industi'v  is  not  willing  to  un- 
dertake hut  that  might  he  in  tlu;  national  in- 
terest. 

B.  Congress  could  establish  a separate  Institute 
of  Biotechnology  as  a funding  agency. 

The  merits  of  a sepai'atc;  institution  lie  in  the 
possibility  of  coordinating  a wide*  range  of  ef- 
forts, all  related  to  hiotechnolog\'.  .Among  pre.s- 
ent  organizations,  biotechnology  and  ap|)lic*d  ge- 
netics cut  across  several  institutes  and  di\  isions 
within  them.  McKlically  oricMiled  r(*seareh  falls 
primarily  under  the  domain  of  the  Natiotial  In- 
stitutes of  Health  (NIH).  Id’, A is  concerned  with 
the  prevention  of  pollution;  w bile  NSt  s etiort  m 
biotechnology  has  hecMi  icstiicted  to  modest 
support  scattered  thi’ough  several  divisions 


Ch.  7 — The  Use  of  Genetically  Engineered  Micro-Organisms  in  the  Environment  • 129 


rhe  creation  of  an  organization  such  as  the  Na- 
tional Technolog)’  Foundation  (H  R.  (S910)  would 
represent  the  kind  of  commitment  to  engineer- 
ing. in  general,  that  currently  does  not  exist. 

C^ompetition  for  funds  \\  ithin  other  agencies 
would  i)e  a\  oided,  since  funding  \\  oukl  now  oc- 
cur at  the  le\  el  of  congressional  appropriations. 
.A  separate  institute,  carrying  the  stamp  of 
(io\ernment  recognition,  would  make  it  clear  to 
the  puhlic  that  this  is  a major  new  area  with 
great  potential.  This  might  foster  greater  aca- 
demic and  commercial  interest  in  hiotechnolog\’ 
and  genetic  engineering. 

On  the  other  hand,  hiotechnolog\'  and  genetic 
engineering  co\  er  such  a broad  range  of  disci- 
plines that  a single  agency  would  over  lap  the 
mandates  of  existing  agencies.  Furthermoi'e,  the 
creation  of  yet  another  agency  carries  w ith  it  all 
the  disadv  antages  of  incr'eased  bui'eauci'acy  and 
competition  for  funds  at  the  agency  level. 

C.  Congress  could  establish  research  centers  in 
universities  to  foster  interdisciplinary  ap- 
proaches to  biotechnology'.  In  addition,  a pro- 
gram of  training  grants  could  be  offered  to 
train  scientists  in  biological  engineering. 

The  successful  use  of  biological  techniques  in 
industry  depends  on  a multidisciplinary  ap- 
proach involving  biochemists,  geneticists,  mi- 
crobiologists, process  engineers,  and  chemists. 
Little  is  now  being  done,  publicly  or  privately, 
to  develop  expertise  in  this  interdisciplinary 
area. 

In  1979,  President  Carter  proposed  the  crea- 
tion of  generic  technology  centers  (useful  to  a 
broad  range  of  industries)  as  one  way  to  stim- 
ulate innovation.  The  centers  would  conduct 
the  kind  of  research  that  an  individual  company 
might  not  consider  cost  effective,  but  that  might 
ultimately  benefit  sev  eral  companies.  Each  cen- 
ter would  be  jointly  funded  by  Government  and 
industry,  with  Government  prov  iding  the  seed 
money  and  industry  carrying  most  of  the  costs 
within  5 years.  If  the  centers  were  established 
at  universities,  startup  costs  could  be  mini- 
mized. _ 

Several  congressional  bills  contain  prov  isions 
for  centers  similar  to  these.  For  example,  on 


October  21,  1980,  President  Carter  signed  into 
law  a bill  (S.  1250)  that  would  establish  Centers 
for  Industrial  Technology  to  foster  research 
links  between  industry  and  universities.  They 
would  he  affiliated  with  a university  or  non- 
profit institution. 

One  or  more  of  these  centers  could  be  specifi- 
cally designated  to  specialize  in  biotechnology. 
In  addition,  training  grants  could  be  used  to 
support  the  education  of  hiotechnologists  at  the 
centers  or  elsewhere.  Currently,  there  is  no  na- 
tionwide training  program  to  train  students  in 
this  discipline.  Education  programs,  especially 
for  the  postgraduate  and  graduate  training  of 
engineers,  could  further  the  idea  of  using  bio- 
logical techniques  to  solve  engineering  prob- 
lems. 

D.  Congress  could  use  ta^  incentives  to  stimulate 
biotechnology'. 

The  tax  laws  could  be  used  to  stimulate  bio- 
technologv'  in  several  ways.  First,  they  could  ex- 
pand the  supply  of  capital  for  small  high-risk 
firms,  which  are  generally  considered  more  in- 
novative than  established  firms,  because  of 
their  willingness  to  undertake  the  risks  of  in- 
novation. \luch  of  the  pioneering  work  in  the 
industrial  application  of  genetic  techniques  has 
been  done  by  such  firms.  By  nature,  they  are 
speculative,  high-risk  investments.  Second,  the 
tax  law  could  provide  special  subsidies  to  new 
high-technology  firms,  which  cannot  use  the 
standard  investment  incentives,  such  as  the  in- 
vestment tax  credit,  because  they  usually  have 
no  taxable  profits  for  the  first  several  years 
against  which  to  apply  the  tax  credit.  Third,  tax 
incentives  could  be  provided  for  both  estab- 
lished and  new  firms  to  make  the  investment  of 
money  for  R&D  more  attractive. 

There  are  a number  of  ways  to  expand  the 
supply  of  venture  capital.  One  is  to  decrease  the 
tax  rate  on  capital  gains  or  the  period  an  asset 
must  be  held  for  it  to  be  considered  a capital 
gain  rather  than  ordinary  income.  This  change 
could  be  limited  to  stocks  in  high-technology 
firms  in  order  to  focus  its  impact  and  minimize 
revenue  loss.  Other  options  involving  the  stock 
of  high-technology  companies  are;  a tax  credit 
to  the  investor  who  purchases  the  stock;  defer- 


130  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


ment  of  capital  gains  taxes  on  the  sales  of  these 
stocks  if  the  proceeds  are  reinvested  into  simi- 
larly qualifying  stock;  and  more  liberal  capital 
loss  provisions. 

In  addition  to  focusing  on  the  supply  of  cap- 
ital, tax  policy  could  attempt  to  directly  increase 
the  profitability  of  potential  growth  companies. 
Since  most  are  not  profitable  for  several  years, 
they  cannot  take  full  advantage  of  the  invest- 
ment tax  credit— or  even  the  provision  for  car- 
rying net  operating  losses  back  3 years  and  for- 
ward 7 years  to  offset  otherwise  taxable  profits. 
Two  proposals  may  remedy  this  situation.  First, 
the  investment  tax  credit  could  be  refundable  to 
the  extent  it  exceeded  any  tax  liability  of  the 
firm.  A preliminary  estimate  of  the  revenue  loss 
for  this  proposal  was  $1  billion  for  1979.  Sec- 
ond, new  companies  could  be  permitted  to 
carry  net  operating  losses  forward  for  10  years. 
This  change  would  give  new  firms  the  same 
number  of  years  over  which  to  deduct  losses  as 
established  firms. 

The  final  type  of  tax  incentive  is  directed  at 
increasing  R&D  expenditures.  Two  major  pro- 
posals would  permit  companies  to  take  tax  cred- 
its on  a certain  percentage  of  their  R&D  ex- 
penses, and  on  contributions  to  universities  for 
research. 

The  R&D  credit  has  been  advocated  for  sev- 
eral reasons.  First,  it  would  increase  the  after- 
tax return  on  R&D  investments,  making  them 
more  attractive.  Second,  it  would  reduce  the 
degree  of  risk  on  such  investments;  with  a 10- 
percent  credit,  the  real  after-tax  expense  of  a $1 
million  investment  is  $900,000.  Finally,  it  would 
give  firms  maximum  flexibility  in  selecting  proj- 
ects for  investment. 

Questions  have  been  raised  about  the  cost  ef- 
fectiveness of  the  credit.  For  calendar  year 
1980,  the  Treasury  Department  estimated  the 
cost  of  a 10-percent  R&D  credit  to  be  $1.9  bil- 
lion. Since  R&D  costs  average  only  10  to  20  per- 
cent of  the  total  cost  of  bringing  a new  product 
or  process  to  the  market,  the  net  reduction  in 
the  cost  of  commercializing  an  invention  would 
be  1 to  2 percent.  Moreover,  the  commercial 
stage  of  innovation  is  thought  to  be  riskier  and 
costlier  than  the  technical  stage.  Another  prob- 


lem is  that  the  credit  may  be  a windfall  for  firms 
that  would  be  investing  in  R&D  anyway.  Finally, 
the  credit  would  subsidize  R&D  devoted  to 
minor  product  changes  or  incremental  improve- 
ments in  addition  to  R&D  directed  to  more  fun- 
damental breakthroughs. 

One  of  the  provisions  of  a pending  congres- 
sional bill  (H.R.  5829)  provides  for  a credit  of  25 
percent  for  incremental  research  expenditures 
above  those  for  a base  period.  By  limiting  the 
credit  to  incremental  expenditures,  the  hill 
would  create  a more  cost-effecti\e  credit,  if 
passed. 

The  final  type  of  tax  credit  would  he  for  cor- 
porate contributions  to  university  research.  The 
Treasury  Department  estimated  that  a 25  per- 
cent credit  for  research  in  all  fields  would  cost 
$40  million  in  1980.  This  credit  would  he  tar- 
geted to  more  fundamental  research  and  not  to 
the  subsidy  of  short-term,  incremental  projects 
that  are  usually  a significant  |)art  of  corpoi’ale 
R&D  budgets. 

E.  Congress  could  improve  the  conditions  under 
which  U.S.  companies  can  collaborate  with 
academic  scientists  and  make  use  of  the  tech- 
nology developed  in  universities  in  whole  or  in 
part  at  the  taxpayer's  expense. 

Developments  in  genetic  engimuM'ing  hav(‘ 
kindled  interest  in  this  oj)tion.  Ne\ei'th(>l(\ss,  the 
Government’s  role  in  fostering  uni\(M\sily-aca- 
demic  interaction  is  far  from  accepted.  Such  a 
role  may  limit  the  flexibility  of  a (^oop(>rali\ c (‘f- 
fort.  At  the  very  least,  disincentives  siu'h  as  pat- 
ent restrictions  could  he  remo\  ('d. 

The  controversy  has  hecMi  summed  up  as  fol- 
lows:' 

At  the  next  level  of  invoIvcMiienl,  iIk'  (io\crn- 
ment  could  identify  [)otential  partners,  and  fa- 
cilitate negotiations.  A more  active;  role*  would 
inv'olve  the  Government's  pieniiling  startup 
funds.  Finally,  the  GovernmeMit  could  he  a third 
partner,  sharing  costs  with  industry  and  the 
university.  In  this  case,  too  laige*  a Government 
role  could  lead  to  Fedeial  interventioti  in  activ 
ities  that  should  he  the  i(*sponsihility  ol  busi- 
ness and  industry. 


'Dennis  I’rager,  (i.  S.  Omenn.  Srirnrr  207  ;)7!t  SK  I eiKti 


Ch.  7 — The  Use  of  Genetically  Engineered  Micro-Organisms  in  the  Environment  • 131 


Certainly  the  Go\  ernment  can  facilitate  com- 
munication: in  the  health  field,  MH,  for  in- 
stance, is  an  effectixe  stimulus  for  contacts 
among  scientists. 


The  possible  ad\  antages  and  disacK  antages  of 
university-industry  interaction  is  illusti'ated  by 
a recent  case  in\  ol\  ing  a plan  by  Harx  ard  L'ni- 
\ ersity  to  collaborate  with  a genetic  engineering 
company.  The  plan  had  called  for  the  establish- 
ment of  a corporation  to  commercialize  the 
results  of  research  being  done  in  the  laboratory 
of  a Harxard  molecular  biologist,  who  would 
ha\e  been  a principal  in  the  firm.  The  Univer- 
sity would  not  ha\e  been  inxoKed  in  financing 
or  managing  the  firm,  which  would  also  ha\e 
been  housed  separately  from  the  campus.  How- 
ever, Harxard  would  ha\e  derixed  substantial 
income  if  the  company  proxed  successful 
through  a gift  of  10  to  15  percent  of  the  equity 
and  a royalty  on  sales.  .After  much  debate 
among  the  Harxard  faculty  and  educators  na- 
tionxvide,  the  administration  decided  not  to  im- 
plement the  plan  because  of  concerns  about 
possible  adx  erse  impacts  on  academic  x alues. 


Proponents  of  such  arrangements  argue  that 
the  unix  ersities  should  reap  some  return  from 
the  commercialization  of  research  conducted 
by  tbeir  staff.  In  addition,  many  universities  are 
pressed  for  money,  and  joint  xentures  or  re- 
search funding  arrangements  xvith  industry 
provide  an  attractixe  source  of  funds  for  re- 
search programs,  especially  xvhen  Federal  sup- 
port may  decline.  In  return,  industry  xvould 
gain  access  to  the  kind  of  fundamental  research 
that  is  the  foundation  for  innovation  and  ap- 
pears to  be  especially  crucial  in  the  field  of  ge- 
netic engineering,  xvhere  the  gap  betxveen  basic 
research  and  product  dexelopment  is  smaller 
than  for  other  fields. 


Opponents  of  these  arrangements,  especially 
ones  inx'olx’ing  significant  interaction  as  in  the 
Harx  ard  plan,  fear  that  the  profit-seeking  goals 
of  industry  may  be  incompatible  xvith  academic 
x alues.  The  folloxx  ing  possible  adverse  impacts, 
among  others,  have  been  articulated:  1)  in- 
crease in  secrecy,  to  the  detriment  of  the  free 
exchange  of  ideas  so  important  in  academia;  2) 
discrimination  by  the  university  in  its  hiring  and 
promotion  policies  in  fax  or  of  those  doing  the 


rexenue-producing  research;  and  3)  distortion 
in  the  direction  of  research  and  in  the  training 
of  graduate  students. 

F.  Congress  could  mandate  support  for  specific 
research  tasks,  such  as  pollution  control  using 
microbes. 

Inxestment  in  creating  microbes  to  degrade 
pollutants  is  sloxv  because  the  potential  market 
is  thought  to  be  small  and  because  of  the  severe 
liability  problems  that  might  arise  from  inten- 
tional release  of  commercially  supplied  mi- 
crobes. 

But  microbes  may  be  useful  in  degrading  in- 
tractable xxaste  and  pollutants.  Genetic  deter- 
minants for  desired  degradation  activities  may 
be  present  in  naturally  occurring  organisms,  or 
scientists  may  haxe  to  combine  genes  from  dif- 
ferent sources  into  a single  organism.  Current 
research,  hoxxexer,  is  limited  to  isolating  orga- 
nisms from  natural  sources  or  from  mutated 
cultures.  More  elaborate  efforts,  involving  re- 
combinant DNA  (rDNA)  techniques  or  other 
forms  of  microbial  genetic  exchange,  will  re- 
quire additional  effort. 

A decision  by  the  Federal  Government  to  sup- 
port research  and  to  reduce  liability  concerns  is 
probably  needed  before  the  potential  of  micro- 
bial control  of  pollution  can  be  realized.  Federal 
actix  ity  might  depend  on  the  results  of  an  eval- 
uation of  the  technical  feasibility  of  microbial 
pollution  control,  xvhich  could  be  made  by 
either  an  interagency  task  force  or  a special 
commission.  If  the  evaluation  is  negative.  Con- 
gress might  elect  to  do  nothing  to  encourage  the 
technology.  If  the  evaluation  is  positive.  Con- 
gress might  select  from  the  following  sub- 
options: 

1.  Initiate  no  research  support  nor  any  Fed- 
eral relief  from  or  limit  on  potential  liabili- 
ty claims.  This  option  would  not  foreclose 
private  commercial  efforts,  but  it  would 
limit  them  because  of  restricted  research 
funds  and  large  liability  questions.  If  suffi- 
ciently large  markets  were  anticipated  or 
found,  the  limitations  would  be  overcome. 

2.  Initiate  research  support  programs.  Re- 
search might  be  directed  at  problems 
posed  by  particular  pollutants  (contract  re- 


132  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


search).  Federal  support  of  biological  re- 
search is  managed  by  several  agencies,  and 
this  course  would  create  few,  if  any,  major 
administrative  problems. 

3.  Guarantee  markets  for  particular  prod- 
ucts. In  addition  to  patent  protection, 
which  would  be  of  little  value  in  the  case  of 
an  organism  purposefully  disseminated 
into  the  environment,  the  Government 
could  offer  to  buy  desirable  microbes.  This 
public  sector  market  might  provide  enough 
incentive  to  research  to  make  Federal  fund- 
ing unnecessary,  or  the  market  incentive 
and  research  support  might  be  used  jointly. 

4.  Fix  a limit  on  liability  and  set  up  liability  in- 
surance, funded  partly  or  wholly  by  tbe 
Government.  This  option  would  reduce  tbe 
financial  risk  for  entrepreneurs  who  ven- 
ture to  clean  up  pollutants  with  microbes. 
Sucb  an  insurance  scheme  would  require 
that  a Federal  agency  (EPA,  for  instance)  be 
satisfied  that  little  risk  was  attendent  in  the 
use  of  the  microbe. 

5.  Arrange  a scheme  to  test  micro-organisms 
for  known  and  anticipated  risks  before 
they  are  released.  The  Federal  Government 
might  have  to  bear  these  costs  as  part  of  a 
research  program. 

6.  Leave  most  efforts  to  industry  and  allow 
each  Government  agency  to  develop  pro- 
grams in  the  fields  of  genetic  engineering 
and  biotechnology  as  it  sees  fit. 


This  option,  currently  the  status  quo,  seems 
to  be  favored  by  some  industry  officials.  If  it  is 
worth  doing,  they  argue,  industry  will  do  it.  To 
a large  extent,  the  availability  of  venture  capital 
in  the  United  States  has  allowed  many  com- 
panies to  pursue  projects  that  are  deemed  prac- 
tical and  economically  important.  The  produc- 
tion of  interferon,  insulin,  ethanol,  ethylene 
glycol,  and  fructose  are  cited  as  examples  of 
successful  applications  that  were  motivated  by 
industry. 

Generic  research,  or  research  that  is  fun- 
damentally useful  to  a broad  range  of  com- 
panies, will  probably  not  be  undertaken  by  any 
one  company.  When  the  payoff  does  not  come 
soon  enough,  the  Government  has  traditionally 
taken  the  responsibility  for  funding  the  work. 
E.g.,  NIH  supported  717  basic  research  projects 
involving  rDNA  in  fiscal  year  1980  at  a cost  of 
$91.5  million.  Similarly,  high-risk  research  with 
high  capital  costs  would  be  likely  targets  for 
Government  support. 

Leaving  all  R&.D  in  industry's  hands  would 
still  produce  major  commercial  successes,  but 
would  not  ensure  the  development  of  generic 
knowledge  or  the  undertaking  of  high-risk  proj- 
ects. 


Part  II 

Agriculture 


Chapter  8— The  Application  of  Genetics  to  Plants 137 

Chapter  9— Advances  in  Reproductive  Biology  and  Their  Effects  on  Animal  Improvement . . . 167 


chapter  8 

The  Application  of 
Genetics  to  Plants 


chapter  8 


Page 

Perspective  on  Plant  Breeding 137 

The  Plant  Breeder’s  Approach  to 

Commercialization  of  New  Varieties 138 

Major  Constraints  on  Crop  Improvement 139 

Genetic  Technologies  as  Breeding  Tools 140 

New  Genetic  Technologies  for  Plant  Breeding  . 141 

Phase  I:  Tissue  Culture  to  Clone  Plants 141 

Phase  II:  Engineering  Changes  to  Alter 
Genetic  Makeup;  Selecting  Desired  Traits.  . 144 
Phase  III:  Regenerating  Whole  Plants  From 

Cells  in  Tissue  Culture 146 

Constraints  on  the  New  Genetic  Technologies  . 149 

Technical  Constraints 149 

Institutional  Constraints 150 

Impacts  on  Generating  New  V'arieties 151 

Examples  of  New  Genetic  Approaches 152 

Selection  of  Plants  for  Metabolic  Efficiency  . . . 152 

Nitrogen  Fixation 152 

Genetic  Variability,  Crop  Vulnerability,  and 

Storage  of  Germplasm 154 

The  Amount  of  Genetic  Erosion  That  Has 

Taken  Place 154 

The  Amount  of  Germplasm  Needed 154 

The  National  Germplasm  System 155 

The  Basis  for  Genetic  Uniformity 157 

Six  Factors  Affecting  Adequate  Management 
of  Genetic  Resources 158 


Page 

Summary 160 

Issues  and  Options— Plants 161 

Technical  Notes 162 

Tables 

Table  No.  Page 

24.  Average  Yield  per  Acre  of  Major  Crops  in 

1930  and  1975 137 

25.  Some  Plants  Propagated  Through  Tissue 

Culture  for  Production  or  Breeding 143 

26.  Representative  List  of  Tissue  Culture 
Programs  of  Commercial  Significance  in 

the  United  States 143 

27.  Gene  Resource  Responsibilities  of  Federal 

Agencies 155 

28.  Estimated  Economic  Rates  of  Return  From 

Germplasm  Accessions 156 

29.  Acreage  and  Farm  Value  of  Major  U.S.  Crops 

and  Extent  to  Which  Small  Numbers  of 
Varieties  Dominate  Crop  Average 157 

Figures 

Figure  No.  Page 

28.  The  Process  of  Plant  Regeneration  From 

Single  Cells  in  Culture 147 

29.  A Model  for  Genetic  Engineering  of  Forest 

Trees 149 


Chapter  8 

The  Application  of  Genetics  to  Plants 


Perspective  on  plant  breeding 


As  primitive  people  moved  from  hunting  and 
gathering  to  tanning,  they  learned  to  identify 
broad  genetic  ti'aits,  selecting  and  sow  ing  seeds 
from  [jlants  that  grew  faster,  proilucetl  larger 
fruit,  or  were  more  resistant  to  pests  and  dis- 
eases. Often,  a single  trait  that  appeared  in  one 
plant  as  a l esult  of  a mutation  (see  Tech.  ,\ote  1, 
p.  162.)  was  selected  and  bred  to  increase  the 
trait's  frequency  in  the  total  crop  population. 

Mendel’s  laws  of  trait  segregation  enabled 
breeders  to  predict  the  outcomes  of  hybridiza- 
tion and  refinements  in  breeding  methods.  (See 
app.  II-.-\.)  Conseciuentlv , thev  achieved  breed- 
ing objectives  faster  and  with  more  precision, 
significantly  increasing  production.  During  the 
past  80  years  classical  applied  genetics  has  been 
responsible  for: 

• increased  yields: 

• ov  ercoming  natural  breeding  barriers; 

• increased  genetic  diversity  for  specific 
uses: 

• e.xpanded  geographical  limits  where  crops 
can  be  grown;  and 

• improv  ed  plant  quality. 

Since  the  beginning  of  the  20th  century,  plant 
breeders  have  helped  increase  the  productivity 
(see  Tech.  Note  2,  p.  162.)  of  many  important 
crops  for  food,  feed,  fiber,  and  pharmaceuticals 
by  successfully  developing  cultivars  (cultivated 
V arieties)  to  fit  specific  environments  and  pro- 
duction practices.  Some  breeding  objectives 
have  met  the  needs  of  the  local  farmer,  while 
other  genetic  improvements  have  been  applied 
worldwide.  The  commercial  development  of 
hybrid  corn  in  the  1920’s  and  1930’s  and  of 
"green  revolution”  wheats  in  the  1950’s  and 
1960's  are  but  two  examples  of  how  plant 
breeding  has  affected  the  supply  of  food  avail- 
able to  the  world  market.  (See  Tech.  Note  3,  p. 
162.)  A comparison  of  av  erage  yields  per  acre  in 


1930  and  UlTv';  in  table  24  gives  a measure  of  the 
contribution  of  genetics.* 

It  is  im[Dossible  to  determine  exactly  to  wbat 
degree  applied  genetics  has  directly  contributed 
to  increases  in  yield,  because  there  have  been 
simultaneous  improvements  in  farm  manage- 
ment, pest  control,  and  cropping  tecbniques 
using  herbicides,  irrigation,  and  fertilizers.  V'ar- 
ious  estimates,  however,  indicate  that  applied 
genetics  has  accounted  for  as  much  as  50  per- 
cent of  harvest  increases  in  this  century.  The 
yield  superiority  of  new  varieties  has  been  a ma- 
jor impetus  to  their  adoption  by  farmers.  Histor- 
ically, the  primary  breeding  objective  bas  been 
to  maintain  and  improve  crop  yields.  Other 


'(;.  r.  Sprague,  O.  K.  .Vlcwander,  and  J.  VV'.  Uudley,  "Plant  Breed- 
ing and  (ienelic  engineering:  A Perspective,”  BioScience  30(1):  17, 
1980. 

Table  24.— Average  Yield  per  Acre  of  Major  Crops 
in  1930  and  1975 


Average  yield  per  acre  Percent 


1930 

1975 

Unit 

increase 

Wheat 

14.2 

30.6 

Bushels 

115 

Rye 

12.4 

22.0 

Bushels 

77 

Rice.  

46.5 

101.0 

Bushels 

117 

Corn 

20.5 

86.2 

Bushels 

320 

Oats 

32.0 

48.1 

Bushels 

50 

Barley 

23.8 

44.0 

Bushels 

85 

Grain  sorghum.  . . . 

10.7 

49.0 

Bushels 

358 

Cotton 

157.1 

453.0 

Pounds 

188 

Sugar  beets 

11.9 

19.3 

Tons 

62 

Sugarcane 

15.5 

37.4 

Tons 

141 

Tobacco 

. . 775.9 

2,011.0 

Pounds 

159 

Peanuts  

. . 649.9 

2,565.0 

Pounds 

295 

Soybeans 

13.4 

28.4 

Bushels 

112 

Snap  beans 

27.9 

37.0 

Cwt 

33 

Potatoes 

61.0 

251.0 

Cwt 

129 

Onions 

Tomatoes: 

159.0 

306.0 

Cwt 

92 

Fresh  market . . . 

61.0 

166.0 

Cwt 

172 

Processing 

4.3 

22.1 

Tons 

413 

Hops 

. . 1,202.0 

1,742.0 

Pounds 

45 

SOURCE:  U.S.  Department  of  Agriculture,  Plant  Genetic  Resources:  Conserva- 
tion and  Use  (Washington,  D.C.:  USDA,  1979). 


137 


138  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


breeding  objectives  are  specific  responses  to  the 
needs  of  local  growers,  to  consumer  demands, 
and  to  the  requirements  of  the  food  processing 
firms  and  marketing  systems. 

Developing  new  varieties  does  the  farmer  lit- 
tle good  unless  they  can  be  integrated  profitably 
into  the  farming  system  either  by  increasing 
yields  and  the  quality  of  crops  or  by  keeping 
costs  down.  The  three  major  goals  of  crop 
breeding  are  often  interrelated.  They  are: 

• to  maintain  or  increase  yields  by  selecting 
varieties  for: 

—pest  (disease)  resistance; 

—drought  resistance; 

—increased  response  to  fertilizers;  and 
—tolerance  to  adverse  soil  conditions. 

• to  increase  the  value  of  the  yield  by  select- 
ing varieties  with  such  traits  as: 

—increased  oil  content; 

—improved  storage  qualities; 

—improved  milling  and  baking  qualities; 
and 

—increased  nutritional  value,  such  as  high- 
er levels  of  proteins. 

• to  reduce  production  costs  by  selecting 
varieties  that: 

—can  be  mechanically  harvested,  reducing 
labor  requirements; 

—require  fewer  chemical  protectants  or 
fertilizers;  and 

—can  be  used  with  minimum  tillage  sys- 
tems, conserving  fuel  or  labor  by  reduc- 
ing the  number  of  cultivation  operations. 

The  plant  breeder's  approach  to 
commercialization  of  new  varieties 

The  commercialization  of  new  varieties 
strongly  depends  on  the  genetic  variability  that 
can  be  selected  and  evaluated.  A typical  plant 
breeding  system  consists  of  six  basic  steps: 

1.  Selecting  the  crop  to  be  bred. 

2.  Identifying  the  breeding  goal. 

3.  Choosing  the  methodological  approach 
needed  to  reach  that  goal. 

4.  Exchanging  genetic  material  by  breeding. 

5.  Evaluating  the  resulting  strain  under  field 
conditions,  and  correcting  any  deficiencies 
in  meeting  the  breeding  goal. 


6.  Producing  the  seed  for  distribution  to  the 
farmer. 

The  responsibilities  for  the  different  breeding 
phases  are  distributed  but  interactive.  In  the 
United  States,  responsibility  for  crop  impro\  e- 
ment  through  plant  breeding  is  shared  by  the 
Federal  and  State  governments,  commercial 
firms,  and  foundations.^  Although  some  specific 
genes  have  been  identified  for  breeding  pro- 
grams, most  improvements  are  due  to  gradual 
selection  for  favorable  combinations  of  genes  in 
superior  lines.  The  ability  to  select  promising 
lines  is  often  more  of  an  art  (in\'ol\  ing  years  of 
experience  and  intuition)  than  a science. 

The  plant  breeder’s  approach  is  detei’iiiined 
for  the  most  part  by  the  particular  biological 
characteristics  of  the  crop  being  bred— e.g.,  the 
breeder  may  choose  to  use  a system  of  inbreed- 
ing or  outbreeding,  or  the  two  in  combination, 
as  an  approach  to  controlling  and  manipulating 
genetic  variability.  The  choice  is  influenced  l)v 
whether  a particular  plant  in  question  naturally 
fertilizes  itself  or  is  fertilized  by  a neighboring 
plant.  To  a lesser  degree,  the  breeding  objec- 
tives influence  the  choice  of  methods  and  the  se- 
quence of  breeding  procedures. 

Repeated  cycles  of  self-fertilization  leduce 
the  heterozygosity  in  a plant,  so  that  after  nu- 
merous generations,  the  breeder  has  homozy- 
gous, pure  lines  that  breed  true.  (S(?e  Ti'ch.  Note 
4,  p.  162.)  Cross-fertilization,  on  the*  oth(>r  hand, 
results  in  a new  mixture  of  genes  or  increased 
genetic  variability.  Using  these  two  ap|)i()aches 
in  combination  produces  a hybrid— scnci-al  lin«>s 
are  inbred  for  homozygosity  and  tlu'ii  ci-ossed 
to  produce  a parental  line  of  enhanc(>d  gencMie 
potential.  More  vigorous  hybrids  can  he  se- 
lected for  further  testing,  fhe  (dfeets  of  hyl)rid 
vigor  vary  and  include  earlier  gei  niination,  in- 
creased growth  rate  or  size;,  and  grc’ater  ci'op 
uniformity. 

A second  method  for  exchanging  or  adding 
genes  is  achieved  through  altering  the  number 
of  chromosomes,  or  ploidy  (s(‘e  I'eeh  ,\oi«*  .')  |) 
162.),  of  the  plant.  Sinc(?  chromosomes  are 


^Natioriiil  AcacifMiiv  of  Sciences,  (V»n.ser\.'ifi()/i  iit  f,c/ m/i/.iMn  /), 
sources:  An  Imperative,  U ashinulon  I)  ( I!I7K 


c/7,  fl — The  Application  of  Genetics  to  Plants  • 139 


generally  inherited  in  sets,  plants  whose  ploidy 
is  increased  usnally  gain  full  sets  of  new 
chromosomes.  0\er  one-third  of  domesticated 
species  are  polyploids.^  (ienerally,  crop  im- 
pro\  ement  due  to  increased  ploid\’  corresponds 
to  an  o\erall  enlargement  in  plant  size;  leaxes 
can  he  broader  and  thicker  \\  ith  larger  flow  ers, 
fruits,  or  seeds.  .A  well-known  e.xample  is  the 
cultixated  strawberry,  which  has  four  times 
more  chromosomes  than  the  wild  type,  and  is 
much  fleshier. 

.Another  technique,  called  backcrossing,  can 
improx  e a commercially  superior  x ariety  by  lift- 
ing one  or  more  desirable  traits  from  an  inferior 
one.  Generally,  this  is  accomplished  by  making  a 
series  of  crosses  from  the  inferior  to  the  superi- 
or plant  XX  hile  selecting  for  the  desired  traits  in 
each  successixe  generation.  Self-fertilizing  the 
last  backcrossed  generation  results  in  some 
progenx  that  are  homozygous  for  the  genes  be- 
ing transferred  and  that  are  identical  xxith  the 
superior  xariety  in  all  other  respects.  Single 
gene  resistance  to  plant  pests  and  disease-caus- 
ing agents  has  been  successfully  transferred 
through  backcrossing. 

Major  constraints  on  crop 
improvement 

Txx  o of  the  many  constraints  on  crop  breed- 
ing are  related  to  genetics. 

Many  important  traits  are  determined  by  several 
genes. 

The  genetic  bases  for  improx  ements  in  x'ield 
and  other  characteristics  are  not  completely 
defined,  mainly  because  most  biological  traits, 
such  as  plant  height,  are  caused  by  the  interac- 
tion of  numerous  genes.  Although  many— per- 
haps thousands— of  genes  contribute  to  quan- 
titatix  e traits,  much  x ariation  can  be  explained 
by  a few'  genes  that  haxe  major  impact  on 
the  obserxable  appearance  (phenotype)'^— e.g., 
the  height  of  some  genetic  dwarx'es  in  wheat 
can  be  doubled  by  a single  gene.  Many  other 
genes  contribute  to  the  general  health  of  the 


^W.  J.  C.  Lawrence.  Plant  Breeding  (London:  Edward  Arnold  Ltd., 
1968). 

■•J.  \.  Thompson,  Jr.,  ".Analysis  of  Gene  Number  and  Develop- 
ment in  Polygenic  Systems,"  Stadler  Genetics  Symposium  9:63. 


plant  (such  as  resistance  to  pests  and  diseases), 
although  some  of  their  contributions  are  small 
and  difficult  to  assess.  Fax'orable  combinations 
of  genes  result  in  plants  xx^ell-adapted  to  par- 
ticular groxving  conditions  and  agronomic  prac- 
tices. With  thousands  of  genes  in  a single  plant 
contributing  to  overall  fitness,  the  possible  com- 
binations are  almost  infinite. 

Most  poor  combinations  of  genes  are  elim- 
inated by  selection  of  the  best  progeny;  initially 
faxorable  combinations  are  preserved  and  im- 
prox  ed.  laterally  millions  of  plants  may  be  ex- 
amined each  year  to  find  particularly  favorable 
genotypes  for  development  into  nexv  breeding 
stocks.  Increasingly  sophisticated  field  testing 
procedures,  as  xvell  as  adx  anced  statistical  anal- 
yses, are  noxv  used  to  evaluate  the  success  of 
breeding  efforts.  Oxerall  yield  is  still  the  most 
important  criterion  for  success,  although  con- 
siderable care  is  taken  to  test  stress  tolerance, 
pest  and  disease  resistances,  mechanical  har- 
xestabilitv,  and  consumer  acceptability.  Breed- 
ing programs  xvith  specialized  goals  often  use 
rapid  and  accurate  chemical  procedures  to 
screen  lines  and  progeny  for  improvements. 

Because  the  x igor  of  the  plant  depends  on  the 
interaction  of  many  genes,  it  has  been  difficult 
to  identify  individual  genes  of  physiological 
significance  in  xvhole  plants.  As  a result,  many 
important  genes  have  not  been  mapped  in 
major  crop  species.  There  is  little  doubt  that 
breeders  xvould  select  traits  like  photosynthetic 
efficiency  (the  ability  to  convert  light  to  such 
organic  compounds  as  carbohydrates)  or  miner- 
al uptake  if  the  genes  could  be  identified  and 
manipulated  in  the  same  xvays  that  resistance  is 
selected  for  pathogens. 

It  is  uncertain  how  much  genetic  variation  for  im- 
provement exists. 

Although  the  world’s  germplasm  resources 
have  not  been  completely  exploited,  it  has 
become  more  difficult  for  breeders  to  improve 
many  of  the  highly  developed  varieties  now  in 
use— e.g.,  height  reduction  in  wheat  has  made 
enormous  contributions  to  its  productivity,  but 
further  improvement  on  this  basis  seems  to  be 
limited.®  A parallel  condition  in  the  potato  crop 

®N.  F.  Jensen,  "Limits  to  Growth  in  World  Food  Production,"  Sci- 
ence 201:317,  1978. 


140  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Photo  credit:  U.S.  Department  of  Agriculture 

Bundles  of  wheat  showing  variance  in  height 


was  recognized  by  the  National  Research  Coun- 
cil’s Committee  on  Genetic  Vulnerability  of  Ma- 
jor Crops:® 

If  we  bear  in  mind  the  fairly  recent  origin  of 
modern  potato  varieties  and  that  they  are,  for 
the  most  part,  derived  from  the  survivors  of  the 
late  blight  epidemics  of  the  1840’s  in  Europe  and 
North  America,  it  seems  likely  that  the  genetic 

•^National  Academy  of  Sciences,  Genetic  Vulnerability  of  Major 
Crops,  Washington,  D.C.,  1972. 


base  was  already  somewhat  narrow  by  the  time 
modern  potato  breeding  got  under  way.  The 
five-fold  increase  in  yield  resulting  from  selec- 
tion during  the  last  100  years  of  potato  improve- 
ment has  produced  a group  of  varieties  that  are 
genetically  similar  and  unlikely  to  respond  to 
further  selection  for  yield.  In  the  long  run  re- 
sponse to  selection  for  other  characteristics  is 
also  likely  to  be  limited. 

As  these  examples  indicate,  the  le\  el  of  genetic 
homogeneity  of  some  crops  may  make  selection 
for  higher  yields  in  general  more  difficult. 
Nevertheless,  while  the  genetic  basis  for  o\  erall 
crop  improvement  is  poorly  undei'stood,  refine- 
ments in  plant  breeding  techni(|ues  may  in- 
crease the  potential  for  greater  efficiency  in  the 
transfer  of  genetic  information  for  more  precise 
selection  methods,  and  as  a new  source  of  ge- 
netic variation. 

Besides  these  two  constraints,  othei'  |)i'cs- 
sLires  and  limitations  may  also  affect  crop  pro- 
ductivity; some  are  biological  (see  Itudi.  Not(’  B, 
p.  162.),  requiring  technological  breakthroughs, 
while  others  are  related  to  environmental, 
social,  and  political  factors.  (See  Tech.  Note  7,  p. 
162.)— e.g.,  it  has  been  argued  that  the  agri- 
cultural rate  of  growth  is  declining:  In  1976,  tin? 
U.S.  Department  of  Agriculture  (USDA)  esti- 
mated that  the  total-factor  [yrothictivity  of  U.S. 
agriculture  increased  by  2 percent  per  yeai' 
from  1939  to  I960,  hut  by  only  0.9  pei’cenl  fiom 
the  period  of  1960  to  1970.^ 

'U.S.  Department  of  .Agricultun'.  I•.('()n()mi(>,  Statistics  and  ( «)■ 
operation  Ser\  ic:es,  Agricultural  Product ivity:  I'.t^pamllnfi,  the  l.imils. 
Agriculture  Information  Bulletin  .4:11 . Washington.  I)(  I!I79 


Genetic  technologies  as  breeding  tools 


The  new  technologies  may  provide  potential- 
ly useful  tools,  but  they  must  be  used  in  com- 
bination with  classical  plant  breeding  tech- 
niques to  be  effective.  The  technologies  devel- 
oped for  classical  plant  breeding  and  those  of 
the  new  genetics  are  not  mutually  exclusive, 
they  are  both  tools  for  effectively  manipulating 
genetic  information  through  methods  that  have 
been  adapted  from  genetic  recombination  ob- 
served in  nature.  Plant  breeders  have  many 
techniques  for  artificially  controlling  pollina- 


tion-some are  capable  of  o\ (Mcoming  natural 
harriers  such  as  incompatibility.  Net  (>\en 
though  one  new  technology— proto[)last  lusion 
—allows  breeders  to  o\(M’come  incom[)atihilit\ . 
the  new  plant  must  still  he  selected,  regener- 
ated from  single-c(?ll  culture,  and  evaluated 
under  field  conditions  to  ensure  that  the  genetic 
change  is  stable  and  the  atti  ihutes  ot  the  new 
variety  meet  ('ommereial  re(|uirements  I v.ilu.i 
lion  is  still  the  most  expi'iisiv c and  time-eonsum 
ing  step. 


Ch.  8— The  Application  of  Genetics  to  Plants  • 141 


A'eif'  frenetic  technologies  for 
plant  breeding 

The  recent  breaklliroughs  in  genetic  engi- 
neering permit  the  plant  breeder  to  bypass  tbt' 
various  natural  breeding  barriers  that  have 
limited  control  ot  the  transfer  of  genetic  in- 
j formation.  \\  bile  the  new  technologies  do  not 
' necessai'ily  offer  the  plant  bi'eeder  the  radical 
I changes  that  recombinant  1)\'.\  (rl)\'.\)  technol- 
! ogv  provides  the  microbiologist,  they  will,  in 
theory,  s[)eedup  and  perfect  the  process  of  ge- 
netic refinement. 

The  new  technologies  fall  into  two  catego- 
ries: those  involving  genetic  transformations 
through  cell  fusion,  and  those  involv  ing  the  in- 
sertion or  modification  of  genetic  information 
through  the  cloning  (e.xactly  copving)  ot  t)\'.\ 
and  DN,A  vectors  (transfer  DNA).  Most  genetic 
transformations  require  that  enzymes  digest 
the  plant's  impermeable  cell  wall,  a process  that 
leaves  behind  a cell  without  a wall,  or  a proto- 
plast. f’rotoplasts  can  fuse  w ith  each  other,  as 
well  as  with  other  components  of  cells.  In 
theory,  their  ability  to  do  this  permits  a wider 
e.xchange  of  genetic  information. 

The  approach  e.xploiting  the  new  technol- 
ogies is  usually  a three-phase  program. 

Phase  I.  Isolated  cells  from  a plant  are  estab- 
lished in  tissue  culture  and  kept  aliv  e. 
Phase  If.  Genetic  changes  are  engineered  in 
those  cells  to  alter  the  genetic  makeup 
of  the  plant;  and  desired  traits  are 
selected  at  this  stage,  if  possible. 

Phase  III.  The  regeneration  of  the  altered  single 
cells  is  initiated  so  that  they  grow  into 
entire  plants. 

This  approach  contains  similarities  to  the  genet- 
ic manipulation  of  micro-organisms.  However, 
there  is  one  major  conceptual  difference.  In 
micro-organisms,  the  changes  made  on  the  cel- 
lular level  are  the  goals  of  the  manipulation. 
W ith  crops,  changes  made  on  the  cellular  level 
are  meaningless  unless  they  can  be  reproduced 
in  the  entire  plant.  Therefore,  unless  single  cells 
in  culture  can  be  grown  into  mature  plants  that 
have  the  new,  desired  characteristics— a proce- 
dure which,  at  this  time,  has  had  limited  suc- 
cess—the  benefits  of  genetic  engineering  will 


not  be  widespread.  If  the  harriers  can  be  over- 
come, the  new  technologies  will  offer  a new 
way  to  control  and  direct  the  genetic  character- 
istics of  plants. 

PHASE  I:  TISSPE  CULTURE  TO  CLONE  PLANTS 

Tissue  culture  involves  gi'ovving  cells  from  a 
plant  in  a culture  or  medium  that  will  support 
them  and  keep  them  viable.  It  can  be  started  at 
three  diffei'ent  levels  of  biological  organization: 
with  plant  organs  (functional  units  such  as 
leaves  or  i-oots):*  with  tissues  (functioning  ag- 
gregates of  one  type  of  cell,  such  as  epidermal 
cells  (outermost  layer)  in  a leaf;  and  with  single 
cells,  rissue  cultures  by  themselves  offer  spe- 
cific benefits  to  plant  breeders;  just  as  fermenta- 
tion is  crucial  to  microliial  genetic  technologies, 
tissue  culture  is  basic  to  the  application  of  the 
other  new  genetic  technologies  for  plants. 

The  idea  of  growing  cells  from  higher  plants 
or  animals  and  then  regenerating  entire  plants 
from  these  laboratory-grown  cells  is  not  new. 
However,  a better  scientific  understanding  now 
exists  of  what  is  needed  to  keep  the  plant  parts 
alive. 

In  tissue  culture,  isolated  single  plant  cells  are 
typically  induced  to  undergo  repeated  cell  divi- 
sions in  a broth  or  gel,  the  resulting  amorphous 
cell  clump  Is  known  as  a callus.  It  culture  condi- 
tions are  readjusted  when  the  callus  appears,  its 
cells  can  undergo  further  proliferation.  As  the 
resulting  cells  differentiate  (become  special- 
ized), they  can  grow  into  the  well-organized 
tissues  and  organs  of  a complete  normal  plant. 
The  callus  can  be  further  subcultured,  allowing 
mass  propagation  of  a desired  plant. 

At  this  time,  it  is  not  uncommon  to  produce  as 
many  as  a thousand  plants  from  each  gram  of 
starting  cells;  1 g of  starting  carrot  callus  rou- 
tinely produces  500  plants.  The  ultimate  goal  of 
tissue  culturing  is  to  havm  these  plantlets  placed 
in  regular  soil  so  that  they  can  grow  and  devel- 
op into  fully  functional  mature  plants.  The  com- 
plete cycle  (from  plant  to  cell  to  plant)  permits 
production  of  plants  on  a far  more  massive 
scale,  and  in  a far  shorter  period,  than  is  possi- 
ble by  conventional  means.  (See  table  25  for  a 


‘Also  referred  to  as  organ  culture. 


142  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


Photo  credits:  Flow  Laboratories 
Transfer  of  plantlets  grown  on  agar  to  soil 


cies  basis.  However,  several  conimereial  uses  ol' 
tissue  culture  already  exist.  (See  table  2(t.) 

Storage  of  Germplasm.— lissue  culture 
can  be  used  in  the  long-term  storagtt  of  special- 
ized germplasm,  which  in\ol\(ts  tret'zing  c(*lls 
and  types  of  shoots.  The  culturt?  pro\  idtts  stable 
genetic  material,  reduces  storage?  space,  and 
decreases  maintenance  costs. 

Carrot  tissues  ha\e  been  frozen  in  litiuid  ni- 
trogen, thawed  2 years  later,  and  [(‘generated 
into  normal  plants.  Ibis  techni(|ue  has  also 
proved  successful  with  morning  glories,  syca- 
mores, potatoes,  and  carnations.  Cc’nerally.  the 
technique  is  most  useful  for  plant  material  that 
is  vegetatively  propagated,  although  if  it  can  h(> 
generally  applied  it  could  become  important  for 
other  agriculturally  imj)oi’tant  crops. 

Production  of  Phaniiacciilicals  and 
Other  Chemicals  From  l*Ianl  Cells.— be- 
cause plant  cells  in  culture?  are  similai-  to  micro- 
organisms in  fermentation  systems,  they  can  he 
engineered  to  work  as  "factories  " to  produce 


First  stage  in  plant  tissue  culturing;  inoculation 
of  plant  tissue 


list  of  some  plants  propagated  through  tissue 
culture.) 

Each  of  the  four  stages  of  the  complete 
cycle— establishment  in  culture,  organogenesis, 
plantlet  amplification,  and  reestablishment  in 
soil— requires  precise  biological  environments 
that  have  to  be  determined  on  a species-by-spe- 


Shows  the  gradual  development  of  the  plant  tissue 
on  an  agar  medium 


Ch.  8— The  Application  of  Genetics  to  Plants  • 143 


Table  25.— Some  Plants  Propagated  Through 
Tissue  Culture  for  Production  or  Breeding 


Agriculture  and 
horticulture 

Vegetable  crops 
Asparagus 
Beet 

Brussels  sprouts 

Cauliflower 

Eggplant 

Onion 

Spinach 

Sweet  potato 

Tomato 

Fruit  and  nut  trees 
Almond 
Apple 
Banana 
Coffee 
Date 

Grapefruit 

Lemon 

Olive 

Orange 

Peach 

Fruit  and  berries 
Blackberry 
Grape 
Pineapple 
Strawberry 

Foliage 
Silver  vase 
Begonia 
Cryptanthus 
Dieffenbachia 
Dracaena 
Fiddleleaf 


Pointsettia 
Weeping  fig 
Rubber  plant 
Flowers 
African  violet 
Anthruium 
Chrysanthemum 
Gerbera  daisy 
Gloxinia 
Petunia 
Rose 
Orchid 
Ferns 

Australian  tree  fern 
Boston  fern 
Maidenhair  fern 
Rabbitsfoot  fern 
Staghorn  fern 
Sword  fern 
Bulbs 
Lily 
Daylily 
Easter  lily 
Hyacinth 

Pharmaceutical 

Atropa 

Ginseng 

Pyrethium 

Silviculture  (forestry) 

Douglas  fir 

Pine 

Ouaking  aspen 
Redwood 
Rubber  tree 


SOURCE:  Office  of  Technology  Assessment. 


Photo  credit:  U.S.  Department  ol  Agricutiure 

Seed  samples  being  withdrawn  from  a tank  of  liquid 
nitrogen  where  they  had  been  stored  at  - 190°  C for 
6 months.  In  addition  to  testing  these  seeds  for  retained 
germination  potential,  some  will  be  grown  into  fully  mature 
plants  to  determine  if  any  genetic  changes  occurred 
during  storage 


Table  26.— Representative  List  of  Tissue  Culture  Programs  of  Commercial  Significance  in  the  United  States 


Industry 


Application 


Economic  benefits 


Asparagus  industry Rapid  multiplication  of  seed  stock  Improved  productivity,  earliness,  and  spear  quality 


Chemical  and  pharmaceutical . Biosynthesis  of  chemicals 

Propagation  of  medicinal  plants 

Citrus  industry Virus  elimination 

Coffee  industry Disease  resistance  breeding 

Land  reclamation Mass  propagation 

Ornamental  horticulture Mass  propagation 


Pineapple  industry Mass  propagation 

Strawberry  industry Mass  propagation 


Reduced  production  costs 
High  volumes  of  plants  for  planting 
Improved  quality,  high  productivity 
Disease  resistance 

Availability  of  select  clones  of  wild  species  for  revegetation 

Reduced  costs  of  certain  species 

Virus  elimination  of  certain  species 

Introduction  of  new  selections 

Increased  volumes  of  difficult  selections 

Improved  quality  in  higher  volumes 

Rapid  introduction  of  new  strains 


SOURCE:  Office  of  Technology  Assessment. 


144  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


plant  products  or  byproducts.  In  recent  years, 
economic  benefits  have  been  achieved  from  the 
production  of  plant  constituents  through  cell 
culture.  Among  those  currently  produced  com- 
mercially are  camptothecin  (an  alkaloid  with 
antitumor  and  antileukemic  activity),  proteinase 
inhibitors  (such  as  heparin),  and  antiviral  sub- 
stances. Flavorings,  oils,  other  medicinals,  and 
insecticides  will  also  probably  be  extracted  from 
the  cells. 

The  vinca  alkaloids— vincristine  and  vin- 
blastine, for  instance— are  major  chemothera- 
peutic agents  in  the  treatment  of  leukemias  and 
lymphomas.  They  are  derived  from  the  leaves 
of  the  Madagascar  periwinkle  [Catharanthus 
roseus).  Over  2,000  kilograms  (kg)  of  leaves  are 
required  for  the  production  of  every  gram  of 
vinca  alkaloid  at  a cost  of  about  $250/g.  Plant 
cells  have  recently  been  isolated  from  the  peri- 
winkle, immobilized,  and  placed  in  culture.  This 
culture  of  cells  not  only  continues  to  synthesize 
alkaloids  at  high  rates,  but  even  secretes  the  ma- 
terial directly  into  the  culture  medium  instead 
of  accumulating  it  within  the  cell,  thus  remov- 
ing the  need  for  extensive  extraction  pro- 
cedures. 

Similarly,  cells  from  the  Cowage  velvetbean 
are  currently  being  cultured  in  Japan  as  a 
source  of  L-Dopa,  an  important  drug  in  the 
treatment  of  Parkinson’s  disease.  Cells  from  the 
opium  poppy  synthesize  both  the  plant’s  normal 
alkaloids  in  culture  and,  apparently,  some  alka- 
loids that  have  not  as  yet  been  found  in  extracts 
from  the  whole  plant. 

Another  pharmaceutical,  diosgenein,  is  the 
major  raw  material  for  the  production  of  corti- 
costeroids and  sex  steroids  like  the  estrogens 
and  progestins  used  in  birth  control  pill.  The 
large  tuberous  roots  of  its  plant  source, 
Dioscorea,  are  still  collected  for  this  purpose  in 
the  jungles  of  Central  America,  but  its  cells  have 
been  cultured  in  the  laboratory. 

Other  plant  products,  from  flavorings  and 
oils  to  insecticides,  industrial  organic  chemicals, 
and  sweeteners,  are  also  beginning  to  be  de- 
rived from  plants  in  cell-cultures.  Glycyrrhiza, 
the  nonnutritive  sweetener  of  licorice,  has  been 
produced  in  cultures  of  Glycyrrhiza  glabra,  and 


anthraquinones,  which  are  used  as  dye  bases, 
accumulate  in  copious  amounts  over  several 
weeks  in  cultures  of  the  mulberry,  Morinda  citri- 
folia. 

PHASE  II:  ENGINEERING  CHANGES  TO  ALTER 
GENETIC  MAKECP;  SELECTING  DESIRED  TRAITS 

The  second  phase  of  the  cycle  inxoKes  the 
genetic  manipulation  of  cells  in  tissue  culture, 
followed  by  the  selection  of  desired  traits. 
Tissue  culturing,  in  combination  with  the  new 
genetic  tools,  could  allow  the  insertion  of  new 
genetic  information  directly  into  plant  cells. 
Several  approaches  to  exchanging  genetic  infoi’- 
mation  through  new  engineering  technologies 
exist: 

• culturing  plant  sex  cells  and  embryos; 

• protoplast  fusion;  and 

• transfer  by  DNA  clones  and  foreign  \ec- 
tors. 

These  are  then  followed  by: 

• screening  for  desired  traits. 

Culturing^  Plant  Cells  and  Embryos.— 

Culturing  the  plant’s  sex  cells— the  ogg  from  the 
ovary  and  the  pollen  from  the  anther  (pollen- 
secreting  organ)— can  inci’ease  the  (d'ficitMicy  of 
creating  pure  plant  lines  for  breeding.  Since  sex 
cells  contain  only  a single  set  of  unpaiiH'd 
chromosomes  per  cell,  plantlets  derived  from 
them  also  contain  only  a single  set.  I'hus,  any 
genetic  change  will  heconu;  ap[)arent  in  the  re- 
generated plant,  because  a second  paired  gene 
cannot  mask  its  effect.  I,ai’g(>  numheis  of  hap- 
loid plants  (cells  contain  half  the  normal  num- 
ber of  chromosomes)  haw.  been  |)i  {)duc«‘d  lor 
more  than  20  sp(!cies.  Sim|)l(?  treatment  with 
the  chemical,  colchicine,  can  usually  induc»' 
them  to  du{)licate  their  genomes  lhaploid  .set  of 
chromosomes)— resulting  in  fully  normal,  dip- 
loid plants.  The  only  major  crop  that  has  been 
bred  by  this  technitiue  is  the  aspai  agus  " 

If  the  remaining  technical  harrieis  can  he 
overcome,  the  techni(|ue  can  he  used  to  en- 
hance the  selection  of  ('lite  trees  and  to  create 
hybrids  of  important  crops.  Although  still 


"J.  C.  I'orrev,  "Cvtiulidci  ciiliiilion  in  ( iiltni  rd  ( i-IU  .mil  I 
HortSciencp  12(2):  1 3«.  1!)77 


Ch.  8— The  Application  of  Genetics  to  Plants  • 145 


pi  imarily  experimental,  sueeesst’ul  plant  sex-cell 
cultures  ha\e  l)een  achie\ecl  lor  a \arietv  of 
im[)oi’tant  culti\ars,  including  rice,  tobacco, 
wheat,  hai'lev,  oats,  sorghum,  and  tomato,  llou- 
e\ei',  because  th(>  t(‘chni(|ue  can  lead  to  hizai'i'e 
unstable  chromosomal  ar’rangements,  it  has  had 
few  applications. 

Kmhryo  culturi's  ha\ c been  used  to  g(M'mi- 
nate,  in  \itro,  those  iMiihryos  that  might  not 
otherwise  sur\  i\  e because  of  basic  incom[)atihil- 
ities,  especialh  wlien  plants  from  different 
genei'a  are  crossed.  Kmhi'yos  may  function  as 
starting  material  in  tissue  cultuia'  s\stems  re- 
(|uiring  jmcnile  material.  They  are  being  used 
to  speed  up  germination  in  such  sjjecies  as  oil 
[)alms.  v\hich  take  u[)  to  2 years  to  g(>rminate 
under  natural  coiulitions. 

Protoplast  I'usioii.— In  proto[)last  fusion, 
either  two  entire  protoplasts  are  brought  to- 
gether, or  a single  protoplast  is  joined  to  cell 
components— or  organelles— from  a second  pro- 
toplast. When  the  com[)onents  are  mixed  under 
the  right  conditions,  they  fuse  to  form  a single 
hybrid  cell.  I'he  hv  hrids  can  he  induced  to  pro- 
liferate and  to  regenerate  cell  walls.  The  func- 
tional plant  cell  that  results  may  often  he 
cultured  fui  ther  and  regenerated  into  an  entire 
plant— one  that  contains  a combination  of  genet- 
ic material  from  both  starting  plant  cell  progeni- 
tors. \\  hen  protoplasts  are  induced  to  fuse,  they 
can,  in  theor\',  exchange  genetic  information 
w ithout  the  restriction  of  natural  breeding  har- 
riers. ,At  present,  protoplast  fusion  still  has 
many  limitations,  mainly  due  to  the  instability  of 
chromosome  pairing. 

Organelles  are  small,  specialized  components 
within  the  cell,  such  as  chloroplasts  and  mito- 
chondria. Some  organelles,  called  plastids,  carry 
their  own  autonomously  replicating  genes,  as  a 
result,  they  may  hold  promise  for  gene  transfer 
and  for  carrying  new  genetic  information  into 
protoplasts  in  cultures,  or  possibly  for  influenc- 
ing the  functions  of  genes  in  the  cell  nucleus. 
(See  Tech.  Note  8,  p.  163.) 

The  feasibiliU'  of  protoplast  fusion  has  been 
borne  out  in  recent  work  with  tobacco — a plant 
that  seems  particularly  amenable  to  manipula- 
tion in  culture.  ,An  albino  mutant  of  Nicotiana 


tahacum  was  fused  with  a \arietv  of  a sexually 
incompatible  Nicotiana  species.  The  resultant 
hybrids  were  easily  recognized  by  their  inter- 
mediate light  green  color.  They  ha\  e now  been 
rt^genei'ated  into  adult  plants,  and  are  currently 
being  used  as  a promising  source  of  hornvvorm 
resistance  in  tobacco  plants. 

Iranslfer  by  DIVA  Clontis  and  Foreign 
A'eetors.— Hecomhinant  DNA  technology 
makes  possible  the  selection  and  production  of 
moi-e  copies  (amplification)  of  specific  DNA 
segments.  Se\  eral  basic  approaches  exist.  In  the 
"shotgun”  appi'oach,  the  whole  plant  genome  is 
cut  by  one  or  moi'e  of  the  commercially  avail- 
able restriction  enzymes.  The  DNA  to  he  trans- 
ferred is  then  attached  to  a plasmid  or  phage, 
w hich  carries  genetic  infoi-mation  into  the  plant 
cell.— E.g.,  a gene  coding  for  a protein  (zein)  that 
is  a major  component  of  corn  seeds  has  been 
spliced  into  plasmids  and  cloned  in  micro-orga- 
tiisms.  It  is  hoped  that  the  zein-gene  sequence 
can  he  modified  through  this  approach  to  in- 
crease the  nutritional  quality  of  corn  protein 
before  it  is  reintroduced  into  the  corn  plant. 

f’oreign  \ectors  are  nonplant  materials  (vi- 
ruses and  bacterial  plasmids)  that  can  he  used  to 
transfer  DNA  into  higher  plant  cells.  Trans- 
formation through  foreign  vectors  might  im- 
prove plant  varieties  or,  by  amplifying  the  de- 
sired DNA  sequence,  make  it  easier  to  recover  a 
cell  product  from  culture.  In  addition,  methods 
have  been  discovered  that  eliminate  the  foreign 
DN,A  from  the  transformed  mixture,  leaving 
only  the  desired  gene  in  the  transformed  plant. 
The  most  promising  vector  so  far  seems  to  be 
the  tumor-inducing  (Ti)  plasmid  carried  by 
Agrobacterium  tumefaciens.  This  bacterium 
causes  tumorous  growths  around  the  root 
crow  ns  of  plants.  It  infects  one  major  group  of 
plants— the  dicots  (such  as  peas  and  beans),  so- 
called  because  their  germinating  seeds  initially 
sprout  double  leaves.  Its  virulence  is  due  to  the 
Ti  plasmid,  which,  when  it  is  transferred  to 
plant  cells,  induces  tumors.  Once  inside  the  cell, 
a smaller  segment  of  the  Ti  plasmid,  called  T- 
DNA,  is  actually  incorporated  into  the  recipient 
plant  cell’s  chromosomes.  It  is  carried  in  this 
form,  replicating  right  along  with  the  rest  of  the 


146  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animais 


chromosomal  DNA  as  plant  cell  proliferation 
proceeds.  Researchers  have  been  wondering 
whether  new  genetic  material  for  plant  im- 
provement can  be  inserted  into  the  T-DNA 
region  and  carried  into  plant  cell  chromosomes 
in  functional  form. 

Adding  foreign  genetic  material  to  the  T-DNA 
region  has  proved  successful  in  several  ex- 
periments. Furthermore,  it  has  been  found  that 
one  type  of  plant  tumor  cell  that  contains 
mutagenized  T-DNA  can  be  regenerated  into  a 
complete  plant.  This  new  discovery  supports 
the  use  of  the  Agrobacterium  system  as  a model 
for  the  introduction  of  foreign  genes  into  the 
single  cells  of  higher  plants. 

Many  unanswered  questions  remain  before 
Agrobacterium  becomes  a useful  vector  for 
plant  breeding.  Considerable  controversy  exists 
about  exactly  where  the  Ti  plasmid  integrates 
into  the  host  plant  chromosomes;  some  inser- 
tions might  disrupt  plant  genes  required  for 
growth.  In  addition,  these  transformations  may 
not  be  genetically  stable  in  recipient  plants; 
there  is  evidence  that  the  progeny  of  Ti-plasmid- 
containing  plants  do  not  retain  copies  of  the  Ti 
sequence.  Finally,  Agrobacterium  does  not  read- 
ily infect  monocots  (a  second  group  of  plants), 
which  limits  its  use  for  major  grain  crops. 

Another  promising  vector  is  the  cauliflower 
mosaic  virus  (CaMV).  Since  none  of  the  known 
plant  DNA  viruses  has  ever  been  found  in  plant 
nuclear  DNA,  CaMV  may  be  used  as  a vector  for 
introducing  genetic  information  into  plant 
cytoplasm.  Although  studies  of  the  structural 
organization,  transcription,  and  translation  of 
the  CaMV  are  being  undertaken,  information 
available  today  suggests  that  the  system  needs 
further  evaluation  before  it  can  be  considered 
an  alternative  to  the  Agrobacterium  system. 

Although  work  remains  to  be  done  on  Ti- 
plasmid  and  CaMV  genetic  mechanisms,  these 
systems  have  enormous  potential.  Most  immedi- 
ately, they  offer  ways  of  examining  basic  mech- 
anisms of  differentiation  and  genetic  regulation 
and  of  delineating  the  organization  of  the 
genome  within  the  higher  plant  cell.  If  this  can 
be  accomplished,  the  systems  may  provide  a 
way  of  incorporating  complex  genetic  traits  into 
whole  plants  in  stable  and  lasting  form. 


Screening  for  Desired  Traits.— The  bene- 
fits of  any  genetic  alteration  will  be  realized 
only  if  they  are  combined  with  an  adequate  svs- 
tem  of  selection  to  recover  the  desired  traits.  In 
some  cases,  selection  pressures  can  be  useful  in 
recovery.®  The  toxin  from  plant  pathogens,  foi’ 
example,  can  help  to  identify  disease  resistance 
in  plants  by  killing  those  that  are  not  resistant. 
So  far,  this  method  has  been  limited  to  identify- 
ing toxins  excreted  by  bacteria  or  fungi  and 
their  analog;  after  sugarcane  calluses  were  ex- 
posed to  toxins  of  leaf  blight,  the  resistant  lines 
that  survived  were  then  used  to  dexelop  new 
commercial  varieties.  In  theory,  however,  it  is 
possible  to  select  for  many  important  traits. 
Tissue  culture  breeding  for  resistance  to  salts, 
herbicides,  high  or  low  temperatures,  drought, 
and  new  varieties  that  are  more  responsive  to 
fertilizers  is  currently  under  study. 

Five  basic  problems  must  he  overcome  hefor(> 
any  selected  trait  can  he  considei’ed  beneficial 
(see  figure  28): 

• the  trait  itself  must  he  identifieil: 

• a selection  scheme  must  he  found  to  iden- 
tify cells  with  altered  prop(M’ti('s: 

• the  properties  must  |)rove  to  he  du(*  to  ge- 
netic changes; 

• cells  with  altered  properties  must  cotif(*r 
similar  properties  on  the  vv  hole  plant:  and 

• the  alteration  must  not  adwr.selv  affe('t 
such  commerciallv  important  charactei  is- 
tics  as  yield. 

While  initial  scr(!ens  inv olv  ing  cells  are  easier 
to  carry  out  than  sci’eening  tests  of  entire 
plants,  tolerance?  at  the?  ('(‘Ilular  level  must  he 
confirmed  by  inoculations  of  the  mature  plants 
with  the  actual  pathog(*n  under  field  conditions 

PHASE  III:  HE«E\EKATI\(;  UHOI.E  PEAMS 
FROM  CELLS  I\  TISSl'E  Cl  I-'HIRI: 

New  methods  are  hiMiig  develo|)ed  to: 

• increase  the  speed  with  which  (Tops  are 
multiplied  through  mass  propagation,  and 

• create  and  maintain  disease-tree  plants 

Mass  Propagalioii.— The  greatest  single 
use  of  tissue  culture  systems  to  date  has  heen 
for  mass  propagation,  to  (*stahlish  selected 

®J.  K.  Shepard,  I).  Hidiii-v,  atui  I Shahiii  I’dlaln  I’nilupl.i  i i' 
Crop  Improvi^menl.  " .Science  20K  17  1!1K(I 


Ch.8— The  Application  of  Genetics  to  Plants  • 147 


Figure  28.— The  Process  of  Plant  Regeneration  From  Single  Cells  in  Culture 


Desired  plant 


Leaf 


Virus-free 


Field  performance  tests 


Cell  multiplication 


Cell  wall 
removal 


Tissue 


Exposure  to 
selection  pressure 
e g.,  high  salt 
concentration 

Roots  and 
shoots 


Surviving  cells 
go  on  to  form  callus 


-*■  Root-promoting 
hormones 


The  process  of  plant  propagation  from  single  cells  in  culture  can  produce  plants  with  selected  characteristics.  These  selec- 
tions must  be  tested  in  the  field  to  evaluate  their  performance. 


SOURCE:  Office  of  Technology  Assessment. 


Photo  credit:  Plant  Resources  Institute 


Multiplying  shoots  of  jojoba  plant  in  tissue  culture  on  a 
petri  dish.  These  plants  may  potentially  be  selected  for 
higher  oil  content 

culture  because  of  the  increased  speed  with 
sources  of  impro\ed  seed  or  cutting  material. 
(See  table  26.)  In  some  cases,  producing  plants 
bv  other  means  is  simply  not  economically  com- 
petitive. A classic  example  is  the  Boston  fern, 
which,  while  it  is  easy  to  propagate  from  runner 
tips,  is  commercially  propagated  through  tissue 


which  it  multiplies  and  the  reduced  costs  of 
stock  plant  maintenance.  A tissue  culture  stock 
of  only  2 square  feet  (ft^)  can  produce  20,000 
plants  per  month.’® 

Currently,  mass  production  of  such  cultivars 
as  strawberries  (see  Tech.  Note  9,  p.  163.), 
asparagus,  oil  palms,  and  pineapples  is  being 
carried  out  through  plant  tissue  cultures.”  Very 
recently,  alfalfa  was  propagated  in  the  same 
wav,  giv'ing  rise  to  over  200,000  plants,  several 
thousand  of  w'hich  are  currently  being  tested  in 
field  trials.  Also,  1,300  oil  palms,  selected  for 
high  yield  and  disease  resistance,  are  being 
tested*^  in  Malaysia. Other  crops  not  produced 
by  this  method  but  for  which  cell  culture  is  an 
important  source  of  breeding  variation  include 


'“D.  P.  Holdgate,  “Propagation  of  Ornamentals  by  Tissue  Cul- 
ture," in  Plant  Cell,  Tissue,  and  Organ  Culture,  J.  Feinert  and  Y.  P.  S. 
Bajaj  (eds.)  (New  York:  Springer-V'erlag,  1977). 

"T.  Murashige,  "Current  Status  of  Plant  Cell  and  Organ  Cul- 
tures," HorfScience  12(2):127,  1977. 

'^“The  Second  Green  Revolution,”  special  report,  Business  Week, 
Aug.  25,  1980. 


148  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


beets,  brussels  sprout,  cauliflower,  tomatoes, 
citrus  fruits,  and  bananas.  Various  horticultural 
plants— such  as  chrysanthemums,  carnations, 
African  violets,  foliage  plants,  and  ferns— are 
also  being  produced  by  in  vitro  techniques. 

Accelerating  propagation  and  selection  in 
culture  is  especially  compelling  for  economical- 
ly important  forest  species  for  which  traditional 
breeding  approaches  take  a century  or  more. 
Trees  that  reach  maturity  within  5 years  re- 
quire approximately  50  years  to  achieve  a useful 
homozygous  strain  for  further  breeding.  Spe- 
cies such  as  the  sequoia,  which  do  not  flower 
until  they  are  15  to  20  years  old,  require  be- 
tween 1 and  2 centuries  before  traits  are  sta- 
bilized and  preliminary  field  trials  are  eval- 


A  plantlet  of  loblolly  pine  grown  in  Weyerhaeuser  Co.’s 
tissue  culture  laboratory.  The  next  step  in  this  procedure 
is  to  transfer  the  plantlet  from  its  sterile  and  humid 
environment  to  the  soil 


uated.  Thus,  tissue  culture  production  of  trees 
has  become  an  area  of  considerable  interest. 
Already,  2,500  tissue-cultured  redwoods  ha\e 
been  grown  under  field  conditions  for  compari- 
son with  regular,  sexually  produced  seedlings. 
(See  app.  II-B.)  Loblolly  pine  and  Douglas  fir  are 
also  being  cultured;  the  numher  of  trees  that 
can  be  grown  from  cells  in  100  liters  (1)  of  media 
in  3 months  are  enough  to  reforest  roughly 
120,000  acres  of  land  at  a 12  x 12  ft  spacing.'*  To 
date,  3,000  tissue-cultured  Douglas  firs  ha\  e ac- 
tually been  planted  in  natural  soil  conditions. 
(See  figure  29.) 

'^D.  J.  Durzan,  "Progress  and  Promise  in  I'oresl  Cetielirs."  in 
Proceedings,  50th  Anniversary  Symposium  Paper.  Srirnrr  :ind 
Technology  . . . The  Cutting  Edge  (.\()pleton.  U is:  Institute  of  Paper 
Chemistry,  1980). 


Photo  i*.  . . • f 

A young  Douglas  fir  free  propagated  4 vcU!-  'Qo  'i,.*-  , 
small  piece  of  seedling  leaf  tissue.  Three  y«  I'  a-;-!'  ■ 
at  the  test-tube  stage  seen  in  the  loblolly  pme  : 'i 


Ch.8 — The  Application  of  Genetics  to  Plants  • 149 


Figure  29.— A Model  for  Genetic  Engineering 
I of  Forest  Trees 

1 


- a.  Selection  of  genetic  material  from  germplasm  bank 
^ b.  Insertion  of  selected  genes  into  protoplasts 

c.  Regeneration  of  cells  from  protoplasts  and 
multiplication  of  cell  clones 

d.  Mass  production  of  embryos  from  cells 

e.  Encapsulation  to  form  ‘seeds’ 

I f.  Field  germination  of  ‘seeds' 

g.  Forests  of  new  trees 

I 

SOURCE;  Office  of  Technology  Assessment. 

Creation  and  Maintenance  of  Disease- 
; Free  Plants.— Cultixars  maintained  through 
i standard  asexual  propagation  over  long  periods 
f often  pick  up  viruses  or  other  harmful  path- 
I ogens,  which  while  they  might  not  necessarily 
kill  the  plants,  may  cause  less  healthy  growth.  A 
plant’s  true  economic  potential  may  be  reached 
I only  if  these  pathogens  are  removed— a task 
, which  culturing  of  a plant’s  meristem  (growing 
I point)  and  subsequent  heat  therapy  can  per- 
‘ form.  Not  all  plants  produced  through  these 
methods  are  \ irus-free,  so  screening  cells  for 
viruses  must  be  done  to  ensure  a pathogen-free 
plant.  In  horticultural  species,  the  adv  antages  of 


vii'us-free  stock  often  appear  as  larger  flowers, 
moi'e  \ igorous  growth,  and  improved  foliage 
(|uality. 

T oday,  \ irus-free  fruit  plants  are  maintained 
and  distributed  from  both  pi'ivate  and  public 
re|)ositories.  Work  of  commercial  importance 
has  been  done  with  such  plants  as  sti'awherries, 
sweet  [)otatoes,  citrus,  freesias,  irises,  rhuharhs, 
gooseberries,  lilies,  hops,  gladiolus,  geraniums, 
and  chrysanthemums.'-*  Over  134  \ irus-free 
potato  cultures  have  also  been  developed  by  tis- 
sue culture.'® 

Constraints  on  the  neiv  genetic 
technologies 

Although  genetic  information  has  been  trans- 
ferred by  vectors  and  proto|)last  fusion,  iio  DNA 
transfoi’iiiations  of  commercial  value  have  yet 
been  performed.  The  constraints  on  the  suc- 
cessful application  of  molecular  genetic  technol- 
ogies are  both  technical  and  institutional. 

TECHMCAL  CONSTRAINTS 

Molecular  engineering  has  been  impeded  by  a 
lack  of  understanding  about  which  genes  would 
he  useful  for  plant  breeding  purposes,  as  well  as 
by  insufficient  knowledge  about  cytogenetics. 
In  addition,  the  available  tools— vectors  and 
mutants— and  methods  for  transforming  plant 
cells  using  purified  DNA  are  still  limited. 

Cells  carrying  traits  important  to  crop  pro- 
ductiv’ity  must  be  identified  after  they  have 
been  genetically  altered.  Even  if  selection  for  an 
identified  trait  is  successful,  it  must  be  dem- 
onstrated that  cells  with  altered  properties  con- 
fer similar  properties  on  tissues,  organs,  and, 
ultimately  on  the  whole  plant,  and  that  the 
genetic  change  does  not  adversely  affect  yield 
or  other  desired  characteristics.  Finally,  only 
limited  success  has  been  achieved  in  regenerat- 
ing whole  plants  from  individual  cells.  While  the 
list  of  plant  species  that  can  be  regenerated 
from  tissue  culture  has  increased  over  the  last  5 
years,  it  includes  mostly  vegetables,  fruit  and 

'“M.  Misawa,  K.  Sakato,  M.  Tanaka,  M.  Havashi,  and  H.  Same- 
jima,  "Production  of  Physiologically  Active  Substances  by  Plant 
Cell  Suspension  Cultures,"  H,  E.  Street  (ed  ),  Tissue  Culture  and 
Plant  Science  (New  York:  Academic  Press,  1974). 

'^Murashige,  op.  cit. 


150  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


nut  trees,  flowers,  and  foliage  crops.  Some  of 
the  most  important  crops— like  wheat,  oats,  and 
barley— have  yet  to  be  regenerated.  In  addition, 
cells  that  form  calluses  in  culture  cannot  always 
be  coaxed  into  forming  embryos,  which  must 
precede  the  formation  of  leaves,  shoots,  and 
roots.  Technical  breakthroughs  have  come  on  a 
species-by-species  basis;  key  technical  discov- 
eries are  not  often  applicable  to  all  plants.  And 
even  when  the  new  technologies  succeed  in 
transferring  genetic  information,  the  changes 
can  be  unstable. 

The  hope  that  protoplast  fusion  would  open 
extensive  avenues  for  gene  transfer  between 
distantly  related  plant  species  has  diminished 
with  the  observation  of  this  instability.  How- 
ever, if  whole  chromosomes  or  chromosome 
fragments  could  be  transferred  in  plants  where 
sexual  hybridization  is  presently  impossible,  the 
possibilities  would  be  enormous. 

INSTITUTIONAL  CONSTRAINTS 

Institutional  constraints  on  molecular  genet- 
ics include  those  in  funding,  in  regulation,  in 
manpower,  and  in  industry. 

Federal  funding  for  plant  molecular  genetics 
in  agriculture  has  come  from  the  National 
Science  Foundation  (NSF)  and  from  USDA.  Re- 
search support  in  USDA  is  channeled  primarily 
through  the  flexible  Competitive  Grants  Pro- 
gram (fiscal  year  1980  budget  of  $15  million)  for 
the  support  of  new  research  directions  in  plant 
biology.  The  panel  on  genetic  mechanisms  (an- 
nual budget  less  than  $4  million)  is  of  particular 
significance  for  developing  new  genetic  technol- 
ogies. The  panel’s  charter  specifically  seeks  pro- 
posals on  novel  genetic  technologies.  The  re- 
maining three  panels  concerned  with  plants— 
nitrogen,  photosynthesis,  and  stress— also  sup- 
port projects  to  define  the  molecular  basis  of 
fundamental  plant  properties.  The  success  of 
the  USDA  Competitive  Grants  Program  is  hard 
to  assess  after  just  2 years  of  operation;  how- 
ever, its  budget  over  the  past  2 years  has  severe- 
ly limited  expansion  of  the  program  into  new 
areas  of  research. 

Some  private  institutions'®  argue  that  the 

•®V.  Walbot,  Past,  Present  and  Future  Trends  in  Crop  Breeding, 
Vol.  II,  Working  Papers,  Impact  of  applied  Genetics,  NTIS,  1981. 


Competitive  Grants  Program  is  shifting  support 
from  ongoing  USDA  programs  to  new  genetics 
research  programs  that  are  not  aimed  at  the 
important  problems  facing  agriculture  today. 
There  is  no  opposition  to  supporting  the  molec- 
ular approaches  as  long  as  they  do  not  come  at 
the  expense  of  traditional  breeding  programs, 
and  as  long  as  both  molecular  biologists  and 
classical  geneticists  working  with  major  croj) 
plants  are  assured  of  enough  support  to  foster 
research  groups  of  sufficient  size. 

At  present,  funds  from  nine  programs  at  the 
NSF— primarily  in  the  Directorate  for  Biological, 
Behavioral,  and  Social  Sciences— support  plant 
research.  The  total  sujiport  for  the  plant  sci- 
ences may  he  as  high  as  $25  million,  of  which 
only  about  $1  million  is  designated  specifically 
for  molecular  genetics. 

The  regulation  of  the  release  of  genetically 
altered  plants  into  the  environment  has  not  had 
much  effect  to  date.  As  of  Nm  emluM'  1980,  only 
one  application  (which  re(|uested  exception 
from  the  NIH  Ciuidelines  (see  ch.  11)  to  releasi' 
rDNA-treated  corn  into  the  einironmenti  has 
been  filed  with  the  Office  of  Recombinant  DN,\ 
Activities  (ORDA).  \Vh(4h(;r  regulation  will  pro- 
duce major  obstacles  is  difficult  to  predict  at 
present.  It  is  also  unclear  whetlKM’  restrictions 
w'ill  be  placed  on  other  geii(4ic  acli\  itii's,  such 
as  protoplast  fusion.  Gurrently,  at  least  one 
other  nation  (New  Zealand)  includes  such  re- 
strictions in  its  guidelines.  It  is  not  clear  how 
much  the  uncertainty  of  possible  ecological  dis- 
ruption and  the  attiMidiMil  liability  coiucrns 
from  intentional  release  of  genetically  engi- 
neered plants  has  pre\(Mit('d  the  industrial  sec- 
tor from  mo\  ing  toward  comnu'rcial  application 
of  the  new'  technology. 

Only  a few  universities  have  e,\prrtisr  in  both 
plant  and  molecular  biolog\’.  In  addition.  onl\  .i 
few'  scientists  work  with  imnlern  molecular 
techni(|ues  related  to  w holi'  plant  problems  .\s 
a result,  a business  firm  could  easily  diwelop  .i 
capability  exceeding  that  at  an\’  indi\idu.il  I .S 
university.  Howener,  building  industrial  lahor.i 
lories  and  hiring  from  the  uni\ ci  sities  could 
easily  deplete?  the?  ex|)erti.s(‘  at  the  uni\ersit\ 
le\el.  V\'ilh  the?  ri'cent  iincstment  acti\it\  in 
bioengineering  firms,  this  tr»*nd  has  aliead\ 


Ch.8 — The  Application  of  Genetics  to  Plants  • 151 


iH’gun:  in  the'  long-run  it  could  ha\  e serious  con- 
se(|uences  tor  the  (|uality  of  uni\  ersity  research. 

Despite  these  consti'aints,  [)i'ogi'ess  in  o\er- 
coining  the  difficulties  is  continuing.  .\t  the 
prestigious  1980  (lordon  Conference,  w here  sci- 
entists meet  tt)  e.xchange  ideas  and  recent  find- 
ings, plant  moleculai'  hiolog\'  was  added  to  the 
list  of  meetings  for  the  first  time.  In  addition, 
four  other  recent  meetings  ha\e  concentrated 
on  plant  molecular  hiolog\  . ''  Up  to  .10  [)ercent 
of  the  par'ticipants  at  these  meetings  came  from 
nonplant-oriented  disciplines  .searching  for  fu- 
ture re.search  topics.  I his  influ.x  of  in\  estigators 
from  other  fields  can  he  expected  to  enrich  the 
\ariet\’  of  appi'oaches  u.sed  to  soKe  the  prob- 
lems of  the  plant  hrt'eder. 

'^Genome  Ort^nnization  and  Espression  in  Plants.  .\  \ I'O  sym- 
posium held  in  Kdinhurf’h.  Srollund.  July  1979:  Gcnrtic  Enf^ineering 
of  Symbiotic  \itmgen  Fixation  and  Conservation  of  Fi.xed  .\itrof^en. 
June  29-July  2.  19S0.  raluH'  City,  ('alit':  ' .Molerular  Biologists  l.<K)k 
at  (ireen  Plants. ' Siyth  Annual  Symposium.  Sept.  29-Oel.  2.  t9H(). 
Heidelberg.  W est  liermany:  anil  Fourth  International  Svmposium 
on  .S'itro^en  Fixation.  Dec.  l-.i.  19«0.  Canberra,  \ustralia. 


Impacts  on  generating  nei 

Progress  in  the  manipulation  of  gene  expres- 
sion in  eukaryotic  (nucleus-containing)  cells, 
which  include  the  cells  of  higher  plants,  has 
been  enormous.  Most  of  the  new  methodologies 
have  been  derhed  from  fruit  flies  and  mam- 
malian tissue  culture  lines:  but  many  should  be 
directly  applicable  to  studies  with  plant  genes. 
There  has  been  great  progress  in  isolating  spe- 
cific RXA  from  plants,  in  cloning  plant  DNA,  and 
in  understanding  more  about  the  organization 
of  plant  genomes.  Techniques  are  available  for 
manipulating  organs,  tissues,  cells,  or  pro- 
toplasts in  culture;  for  selecting  markers;  for 
regenerating  plants;  and  for  testing  the  genetic 
basis  of  novel  traits.  So  far  however,  these 
techniques  are  routine  only  in  a few  species. 
Perfecting  procedures  for  regenerating  single 
cells  into  whole  plants  is  a prerequisite  for  the 
success  of  many  of  the  novel  genetic  technol- 
ogies. In  addition,  work  is  progressing  on 
\ iruses,  the  Ti  plasmid  of  Agrobacterium,  and 
engineered  cloning  vehicles  for  introducing 
DNA  into  plants  in  a directed  fashion.  There 


Finally,  as  a general  rule,  tradeoffs  arise  in 
the  use  of  the  new  technologies  that  may  inter- 
fere with  their  ajjplication.  It  is  impossible  to  get 
something  for  nothing  from  nature— e.g.,  in  ni- 
trogen fixation  the  symbiotic  relationship  bet- 
ween plant  and  micro-organism  requires  ener- 
gy' from  tbe  plant:  screening  for  plants  that  can 
produce  and  transfer  the  end  products  of  pho- 
tosynthesis to  the  nodules  in  the  root  more  effi- 
ciently may  reduce  inorganic  nitrogen  require- 
ments hut  may  also  reduce  the  overall  yield. 
This  was  the  case  for  the  high  lysine  varieties  of 
corn.  (See  Tech.  Note  10,  p.  163.)  Farmers  in  the 
Lhiited  States  tended  to  avoid  them  because  im- 
proving the  protein  quality  reduced  the  yield, 
an  unacceptable  tradeoff  at  the  market  price. 
Thus,  unless  the  genetic  innovation  fits  the  re- 
quii'ements  of  the  total  agricultural  industry, 
potentials  for  crop  improvement  may  not  be 
realized. 


varieties  

have  been  few  demonstrations  in  which  the  in- 
heritance of  a new  trait  was  maintained  over 
several  sexual  generations  in  the  whole  plant. 

Because  new  varieties  have  to  be  tested 
under  different  environmental  conditions  once 
the  problems  of  plant  regeneration  are  over- 
come, it  is  difficult  to  assess  the  specific  impacts 
of  the  new  technologies.— E.g.,  it  is  impossible  to 
determine  at  this  time  whether  technical  and 
biological  barriers  will  ever  be  overcome  for 
regenerating  wheat  from  protoplasts.  Never- 
theless, the  impact  of  genetics  on  the  structure 
of  American  agriculture  can  be  discussed  with 
some  degree  of  confidence. 

Genetic  engineering  can  affect  not  only  what 
crops  can  be  grown,  but  where  and  how  those 
crops  are  cultivated.  Although  it  is  a variable  ii 
production,  it  usually  acts  in  conjunction  with 
other  biological  and  mechanical  innovations, 
whose  deployment  is  governed  by  social,  eco- 
nomic, and  political  factors. 


152  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


Examples  of  netv  genetic  approaches 

The  ways  in  which  the  new  genetic  ap- 
proaches could  aid  modern  agriculture  are 
described  in  the  following  two  examples: 

SELECTION  OF  PLANTS  FOR 
METABOLIC  EFFICIENCY 

Because  terrestrial  plants  are  immobile,  they  live  and  die 
according  to  the  dictates  of  the  soil  and  weather  conditions 
in  which  they  are  planted;  any  environmental  stress  can 
greatly  reduce  their  yield.  The  major  soil  stresses  faced  by 
plants  include  insufficient  soil  nutrients  and  water  or  toxic 
excesses  of  minerals  and  salts.  The  total  land  area  with 
these  conditions  approaches  4 billion  hectares  (ha),  or 
about  30  percent  of  the  land  area  of  the  Earth. 

Traditionally,  through  the  use  of  fertilizers,  lime, 
drainage,  or  freshwater  irrigation,  environments  have 
been  manipulated  to  suit  the  plant.  Modern  genetic  tech- 
nologies might  make  it  easier  to  modify  the  plant  to  suit  the 
environment. 

Many  micro-organisms  and  some  higher  plants  can  tol- 
erate salt  levels  equal  to  or  greater  than  those  of  sea  water. 
While  salt  tolerance  has  been  achieved  in  some  varieties  of 
plants,  the  classical  breeding  process  is  arduous  and  lim- 
ited. If  the  genes  can  be  identified,  the  possibility  of  actual- 
ly transferring  those  for  salt  tolerance  into  plants  makes 
the  adaptation  of  plants  to  high  salt,  semiarid  regions  with 
high  mineral  toxicities  or  deficiencies  a more  feasible  pros- 
pect. In  the  future,  selecting  among  tissue  cultures  for 
metabolic  efficiency  could  become  important.  Tissue 
culture  systems  could  be  used  to  select  cell  lines  for 
resistance  to  salts  and  for  responsiveness  to  low-nutrient 
levels  or  less  fertilizer.  However,  too  little  is  known  about 
the  biochemistry  and  physiology  of  plants  to  allow  a more 
directed  approach  at  this  time.  Chances  for  success  would 
be  increased  with  a better  understanding  of  plant  cell 
biology. 

Such  techniques  could  be  applied  to  agricultural  pro- 
grams in  less  developed  countries,  where,  commonly,  sup- 
plies of  fertilizers  and  lime  are  scarce,  the  potential  for  ir- 
rigation is  small,  and  adequate  support  for  technological 
innovation  is  limited.  In  addition,  the  United  States  itself 
contains  marginal  land  that  could  be  exploited  for  forest 
products  and  biomass.  The  semiarid  lands  of  the  South- 
west, impoverished  land  in  the  Lake  States,  and  reclaimed 
mining  lands  could  become  cost-effective  areas  for  produc- 
tion. 

NITROGEN  FIXATION 

It  has  been  known  since  the  early  1800’s  that  biological 
fixation  of  nitrogen  is  important  to  soil  fertility.  In  fixation, 
micro-organisms,  such  as  the  bacterium  Rhizobium, 
transform  atmospheric  nitrogen  into  a form  that  plants 
can  use.  In  some  cases— e.g.,  with  legumes  this  process  oc- 
curs through  a symbiotic  relationship  between  the  micro- 
organism and  the  plant  in  specialized  nodules  on  the  plant 
roots.  Unfortunately,  the  major  cereal  crops  such  as 


wheat,  corn,  rice,  and  forage  grasses  do  not  have  the 
capacity  to  fix  atmospheric  nitrogen,  thus  are  largely 
dependent  on  chemically  produced  nitrogen  fertilizers. 
Because  of  these  crops,  it  has  been  estimated  that  the 
world  demand  for  nitrogen  fertilizers  will  grow  from  51.4 
million  metric  tonnes  (1979  estimate)  to  144  million  to  180 
million  tonnes  by  the  year  2000.’*  Therefore,  geneticists 
are  looking  into  the  possibility  that  the  genes  for  nitrogen 
fixation  present  in  certain  bacteria  (called  "nif  genes")  can 
be  transferred  to  the  major  crops. 

Laboratory  investigation  has  focused  on  the  molecular 
biology  of  nitrogen  fixation  in  the  free  living  bacterium, 
Klebsiella  pneumoniae.  A cluster  of  15  nif  genes  has  been 
successfully  cloned  onto  bacterial  plasmids  using  rllN.A 
technology.  These  clones  are  being  used  to  study  the 
molecular  regulation  of  nif  gene  expression  and  the 
physical  organization  of  the  nif  genes  on  the  Klebsiella 
chromosome.  In  addition  they  have  aided  the  search  for 
nitrogen  fixation  genes  in  other  bacteria. 

It  is  thought  that  a self-sufficient  package  of  nitrogen- 
fixing genes  evolved  during  the  course  of  plant  adaptation, 
and  that  this  unit  has  been  transferred  in  a functional 
form  to  a variety  of  different  bacterial  spt^cies,  including 
Klebsiella  and  Rhizobium.  If  the  right  IlNA  \ector  can  he 
found,  the  nif  genes  might  he  transferred  from  bacteria  to 
plants.  The  chloroplasts,  the  cauliflower  mosaic  \ irus,  and 
the  Agrobacterium  Ti-plasmid  are  being  in\estigated  as 
possible  vectors. 

The  way  that  Agrubacteria,  in  particular,  infect  cells  is 
similar  to  the  way  Rhizobia  infect  plants  and  form 
nitrogen-fixing  nodules.  In  both  cases,  the  |)hysical  attach- 
ment between  bacterium  and  plant  tissue  is  necessary  for 
successful  infectioti.  In  the  case  of  Agrobarleria,  tumors 
form  when  a segment  of  the  Ti-plasmid  is  ins«*rted  into  the 
nuclear  genome  of  the  |)lant  cell.  Scientists  do  not  yet 
know  exactly  how  a segment  of  tin?  rhizohi.il  genome  is 
transferred  into  the  root  tissue  to  induce  the  formation  of 
nodules;  nevertheless,  it  is  ho|)ed  that  Agrobarleria  u ill  act 
as  vectors  for  the  introduction  and  expression  of  toriMgn 
genes  into  plant  cells,  just  as  Rhizobia  do  naturalK 

Other  researchers  ha\c  hec'ii  iincstigating  the  re- 
quirements for  getting  nif  geni's  to  exjiress  themselves  m 
plants.  Nif  genes  from  Klebsiella  have  alie.idy  been 
transferred  into  common  yeast,  an  organism  that  can  he 
grown  in  environments  without  o.xvgen  Unfortun.itelv 
the  [jresence  of  oxygen  destroys  a majoi'  enzyme  lor 
nitrogen  fixation  and  sevei'ely  limits  the  potential  .ipplic  .i 
tions  in  higher  plants.  Nevei  theless,  it  is  hoped  that  ml 
gene  expression  in  yeast  will  he  applicable  to  higher  pl.iots 

An  approach  that  does  not  invoKi'  genetu  engineering 
uses  improved  Rhizobia  strains  that  .ire  sviiihiotu  with 
.soybeans.  I hrough  selection,  Rhizobia  imit.inis  .ire  being 
found  that  out  perform  the  original  wild  strains  I uriher 


'“F.  Aiisulief  "Biological  ,\ilrogen  I iv.iliotr  .Sii/i/Hirfaig  /’.i/w'n 
World  Food  and  Kulrilion  Sludv  (U  aNhington.  I)  ( \alional  \i  .ul 
emv  of  Sciences,  l!)77l. 


Ch.  8 — The  Application  of  Genetics  to  Plants  • 153 


testing  is  needed  to  determine  whether  the  impro\ement 
can  he  maintained  in  field  trials,  where  the  improved 
strains  must  compete  against  wild-Upe  Rhizobia  already 
present  in  the  soil. 

Another  wa\  to  improve  nitrogen  fi.xation  is  to  select 
plants  that  ha\e  more  efficient  s\'mhiotic  relationships 
with  nitrogen-fi.xing  organisms.  Since  the  biological  proc- 
ess retiuires  a large  amount  of  energ\'  from  the  plant,  it 
may  he  possible  to  select  for  plants  that  are  more  efficient 
in  producing,  and  then  to  transfer  the  end  products  of 
photosynthesis  to  the  nodules  in  the  roots.  .Also  e.xisting 
nitrogen-fi.xing  bacterial  strains  that  can  interact  w ith  crop 
plants  which  do  not  oidinarily  fix  nitrogen  could  be 
searched  for  or  dev  eloped. 

Keducing  the  amount  of  chemically  fixed  nitrogen 
fertilizer— and  the  cost  of  the  natural  gas  prev  iously  used 
in  the  chemical  process— would  he  the  largest  benefit  of 
successfullv  fixing  nitrogen  in  crops.  Knv  ironmental  bene- 
fits. from  the  smaller  amount  of  fertilizer  runoff  into 
water  systems,  would  accrue  as  well.  But  is  it  difficult  to 
predict  w hen  these  w ill  become  reality.  Experts  in  the  field 
disagree:  some  feel  the  breakthrough  is  imminent:  others 
feel  that  it  might  take  sev  eral  decades  to  achiev  e. 

The  refinements  in  breeding  methods  pro- 
vided bv  the  new  technologies  may  allow  major 
crops  to  be  bred  more  and  more  for  specialized 
uses— as  feed  for  specific  animals,  perhaps,  or 
to  conform  to  special  processing  requirements. 
In  addition,  since  the  populations  in  less  de\el- 
oped  countries  suffer  more  often  from  major 
nutritional  deficiencies  than  those  in  industrial- 
ized countries,  a specific  export  market  of  cere- 
al grains  for  human  consumption,  like  wheat 
with  higher  protein  levels,  may  be  developed. 

But  genetic  methods  are  only  the  tools  and 
catalysts  for  the  changes  in  how  society  pro- 
duces its  food;  financial  pressures  and  Federal 
regulation  will  continue  to  direct  their  course. 
E.g.,  the  automation  of  tissue  culture  systems 
will  decrease  the  labor  needed  to  direct  plant 
propagation  and  drastically  reduce  the  cost  per 
plantlet  to  a level  competitive  with  seed  prices 
for  many  crops.  W^hile  such  breakthroughs  may 
increase  the  commercial  applications  of  many 
technologies,  the  effects  of  a displaced  labor 
force  and  cheaper  and  more  efficient  plants  are 
hard  to  predict. 


Although  it  is  difficult  to  make  economic  pro- 
jections, there  are  several  areas  where  genetic 
technologies  w ill  clearly  have  an  impact  if  the 
predicted  breakthroughs  occur: 

• Batch  culture  of  plant  cells  in  automated 
systems  will  he  enhanced  by  the  ability  to 
engineer  and  select  strains  that  produce 
larger  quantities  of  plant  substances,  such 
as  pharmaceutical  drugs. 

• The  technologies  will  allow  development  of 
elite  tree  lines  that  will  greatly  increase 
yield,  both  through  breeding  programs 
similar  to  those  used  for  agricultural  crops 
and  by  ox  ercoming  breeding  barriers  and 
lengthy  breeding  cycles.  Refined  methods 
of  selection  and  hybridization  will  increase 
the  potential  of  short-rotation  forestry, 
which  can  provide  cellulosic  substrates  for 
such  products  as  ethanol  or  methanol. 

• The  biological  efficiency  of  many  economi- 
cally important  crops  wall  increase.  Ad- 
vances will  depend  on  the  ability  of  the 
techniques  to  select  for  whole  plant  charac- 
teristics, such  as  photosynthetic  soil  and 
nutrient  efficiency.’® 

• Besides  narrowing  breeding  goals,  the 
techniques  will  increase  the  potential  for 
faster  improvement  of  underexploited 
plants  with  promising  economic  value. 

For  such  adxances  to  occur,  genetic  factors 
must  be  selected  from  superior  germplasm,  the 
genetic  contributions  must  be  integrated  into 
improv'ed  cultural  practices,  and  the  improved 
varieties  must  be  efficiently  propagated  for 
distribution. 


’’For  the  soybean  and  tomato  crops,  the  research  area  for  im- 
proved biological  efficiency  received  the  highest  allotment  of 
funds  in  fiscal  year  1978.  Total  funding  was  S12.9  million  for  soy- 
beans and  S2.1  million  for  tomatoes.  The  second  largest  category 
to  be  funded  was  control  of  diseases  and  nematodes  of  soybeans  at 
S5.1  million  and  for  tomato  at  SI. 6 million. 


154  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


Genetic  variability;  crop  vulnerability;  and 
storage  of  germplasm  


Successful  plant  breeding  is  based  on  tbe 
amount  of  genetic  diversity  available  for  the  in- 
sertion of  new  genes  into  plants.  Hence,  it  is 
essential  to  have  an  adequate  scientific  under- 
standing of  how  much  genetic  erosion  has  taken 
place  and  how  much  germplasm  is  needed.  Nei- 
ther of  these  questions  can  be  satisfactorily  an- 
swered today. 

The  amount  of  genetic  erosion  that 
has  taken  place 

Most  genetic  diversity  is  being  lost  because  of 
the  displacement  of  vegetation  in  areas  outside 
the  United  States.  The  demand  for  increased 
agricultural  production  is  a principal  pressure 
causing  deforestation  of  tropical  latitudes  (see 
Tech.  Note  11,  p.  163),  zones  that  contain  exten- 
sive genetic  diversity  for  both  plants  and 
animals. 

It  has  been  estimated  that  several  hundred 
plant  species  become  extinct  every  year  and 
that  thousands  of  indigenous  crop  varieties 
(wild  types)  have  already  been  lost.  However,  it 
is  difficult  to  measure  this  loss,  not  only  because 
resources  are  on  foreign  soil  but  because  ero- 
sion must  be  examined  on  a species-by-species 
basis.  In  theory,  an  adequate  evaluation  would 
require  knowledge  of  both  the  quantity  of  di- 
versity within  a species  and  the  breadth  of  that 
diversity;  this  process  has  in  practice,  just  be- 
gun. What  is  known  is  that  the  lost  material  can- 
not be  replaced. 

The  amount  of  germplasm  needed 

Germplasm  is  needed  as  a resource  for  im- 
proving characteristics  of  plants  and  as  a means 
for  guaranteeing  supplies  of  known  plant 
derivatives  and  potential  new  ones.  Even  if 
plant  breeders  adequately  understand  the 
amount  of  germplasm  presently  needed,  it  is  dif- 


ficult to  predict  future  needs.  Because  pests  and 
pathogens  are  constantly  mutating,  there  is 
always  the  possibility  that  some  resistance  w ill 
be  broken  down.  Even  though  genetic  dix  ersity 
can  reduce  the  severity  of  economic  loss,  an  epi- 
demic might  require  the  introduction  of  a new 
resistant  variety.  In  addition,  other  pressures 
will  determine  which  crops  will  lie  grown  for 
food,  fiber,  fuel,  and  pharmaceuticals,  and  how 
they  will  be  cultivated;  genetic  dixersity  x\  ill  he 
fundamental  to  these  innovations. 

Even  if  genetic  needs  can  he  ade(|uately  iden- 
tified, there  is  disagreement  about  how  much 
germplasm  to  collect.  In  the  past,  its  collection  ' 
has  been  guided  by  differences  in  moriihologx’ 
(form  and  structure),  xx  hich  hax  e not  often  been 
directly  correlated  to  breeding  ohjectixes.  I'ui'- 
thermore,  the  extent  to  xx  hich  the  nexx  gcMUMir 
technologies  xvill  affect  genetic  xariahility,  xiil- 
nerability,  or  the  storage  of  germplasm,  has  not 
been  determined.  (See  apji.  Il-A.) 

In  addition  to  its  uses  in  plant  improx cment. 
germplasm  can  prox  ide  both  old  and  nexx  piod- 
ucts.  Recent  interest  in  gioxx  ing  guax  ule  as  a 
source  of  hydrocarbons  (for  ruhhei-,  energ^v 
materials,  etc.)  has  focused  attention  on  plants 
that  may  possibly  he  undei’utilized.  It  has  been 
found  that  past  collections  of  guaxule  gei  iii- 
plasm  haxe  not  been  ade(|uatelx  maintained, 
making  current  genetic  improx cnK'iits  more  dil- 
ficult.  In  addition,  half  of  the  world's  medicitial 
compounds  are  obtained  from  plants:  maintain- 
ing as  many  xarieties  as  possible  would  ensure 
the  ax  ailaliility  of  compounds  known  to  he  use- 
ful, as  xvell  as  ntnv,  and  as  xot  u nd i.s( ox  e red 
compounds— e.g.,  the  (|uinine  drugs  used  in  the 
treatment  of  malaria  xvere  originally  obtained 
from  the  Cinchona  plant,  ,\  USD  \ collection  ol 
superior  gei’mplasm  (‘stahlish(>d  in  1!M0  in 
Guatemala  xvas  not  maintained  \s  a conse-  1 


Ch.  8 — The  Application  of  Genetics  to  Plants  • 155 


quence,  cliffiiiilties  arose  during  the  V ietnam 
War  when  the  new  antimalarial  drugs  became 
less  ett'ectix  e on  resistant  sti  ains  of  the  j)arasite 
I and  natural  quinines  were  oitce  again  used. 

i 

I An  inq)ortant  ilistinction  exists  between  pre- 
I ser\  ing  genetic  rt'sources  in  situ  and  presets  ing 
germplasm  stored  in  repositories.  Although 
I genetic  loss  can  occui'  at  each  location,  evolu- 
tion will  continue  only  in  natural  ecosystems. 
I W ith  better  stoi’age  tet'hni(iues.  seed  loss  and 
genetic  drift"  can  he  kef)t  to  a minimum.  .Nev  er- 
theless, s})ecies  extinction  in  situ  will  continue. 


The  iXational  Germplasm  System 

I'SDA  has  been  responsible  for  collecting  and 
cataloging  seed  (mostly  from  agriculturally  im- 
portant plants)  since  1898.  Vet  it  is  important  to 
realize  that  other  Federal  agencies  also  have 
responsibilities  for  gene  resource  management. 
(See  table  27.)  Over  the  past  century,  over 
440, 000  plant  introductions  from  more  than  150 
expeditions  to  centers  of  crop  diversity  have 
been  cataloged. 

The  expeditions  were  needed  because  the 
United  States  is  gene  poor.  The  economically  im- 


Table  27.— Gene  Resource  Responsibilities  of  Federal  Agencies 


Type  of  ecosystems 
under  Federal 

Agency ownership/control 

i U.S.  Department  of  Agriculture 
Animal  & Plant  Health  Inspection 
Service — 

Forest  Service Forestlands  and 

rangelands  (U.S. 
National  Forest) 

Science  & Education  Administration  .— 

! Soil  Conservation  Service — 

Department  of  Commerce 
National  Oceanic  & Atmospheric 

Administration Oceans — between  3 

I and  200  miles  off 

the  U.S.  coasts 

Department  of  Energy — 

Department  of  Health  & Human  Services 
National  Institutes  of  Health — 


Responsibilities 


Controls  insect  and  disease  problems  of  commercially 
valuable  animals  and  plants. 

Manages  forestland  and  rangeland  living  resources  for 
production. 

Develops  animal  breeds,  crop  varieties,  and  microbial  strains. 
Manages  a system  for  conserving  crop  gene  resources. 

Develops  plant  varieties  suitable  for  reducing  soil  erosion  and 
other  problems. 


Manages  marine  fisheries. 


Develops  new  energy  sources  from  biomass. 

Utilizes  animals,  plants,  and  micro-organisms  in  medical 
research. 


Department  of  the  Interior 

Bureau  of  Land  Management Forestlands, 

rangelands,  and 
deserts 

Fish  & Wildlife  Service Broad  range  of 

habitats,  including 
oceans  up  to  3 miles 
off  U.S.  coasts 

National  Park  Service Forestlands, 

rangelands,  and 
deserts  (U.S. 
National  Parks) 

Department  of  State — 


(Agency  for  International  Development . . — 

Environmental  Protection  Agency — 

National  Science  Foundation — 

SOURCE:  David  Kapton,  National  Association  for  Gene  Resource  Conservation. 


Manages  forest,  range,  and  desert  living  resources  for 
production. 

Manages  game  animals,  including  fish,  birds,  and  mammals. 


Conserves  forestland,  rangeland,  and  desert-living  resources. 


Concerned  with  international  relations  regarding  gene 
resources. 

Assists  in  the  development  of  industries  in  other  countries 
including  their  agriculture,  forestry,  and  fisheries. 

Regulates  and  monitors  pollution. 

Provides  funding  for  genetic  stock  collections  and  for  research 
related  to  gene  resource  conservation. 


156  • Impacts  of  Applied  Genetics— Micro-Organisms,  Piants,  and  Animais 


portant  food  plants  indigenous  to  the  continen- 
tal United  States  are  limited  to  the  sunflower, 
cranberry,  blueberry,  strawberry,  and  pecan. 
The  centers  of  genetic  diversity,  found  mostly  in 
tropical  latitudes  around  the  world,  are  be- 
lieved to  be  the  areas  where  progenitors  of  ma- 
jor crop  plants  originated.  Today,  they  contain 
genetic  diversity  that  can  be  used  for  plant  im- 
provement. 

It  is  difficult  to  estimate  the  financial  return 
from  the  germplasm  that  has  been  collected, 
but  its  impact  on  the  breeding  system  has  been 
substantial.  A wild  melon  collected  in  India,  for 
instance,  was  the  source  of  resistance  to  pow- 
dery mildew  and  prevented  the  destruction  of 
California  melons.  A seemingly  useless  wheat 
strain  from  Turkey— thin-stalked,  highly  sus- 
ceptible to  red  rust,  and  with  poor  milling  prop- 
erties—was  the  source  of  genetic  resistance  to 
stripe  rust  when  it  became  a problem  in  the 
Pacific  Northwest.  Similarly,  a Peruvian  species 
contributed  "ripe  rot”  resistance  to  American 
pepper  plants,  while  a Korean  cucumber  strain 
provided  high-yield  production  of  hybrid  cu- 
cumber seed  for  U.S.  farmers.  And  a gene  for 
resistance  to  Northern  corn  blight  transferred 
to  Corn  Belt  hybrids  has  resulted  in  an  esti- 
mated savings  of  30  to  50  bushels  (bu)  per  acre, 
with  a seasonal  value  in  excess  of  $200  million.^® 
(See  table  28.) 

The  effort  to  store  and  evaluate  this  collected 
germplasm  was  promoted  by  the  Agricultural 
Marketing  Act  of  1946,  which  authorized  re- 
gional and  interregional  plant  introduction  sta- 
tions (National  Seed  Storage  Centers)  run  coop- 
eratively by  both  Federal  and  State  Govern- 
ments. The  federally  controlled  National  Seed 
Storage  Laboratory  in  Fort  Collins,  Colo.,  was 
established  in  1958  to  provide  permanent  stor- 
age for  seed.  In  the  1970’s,  it  was  recognized 
that  the  system  should  include  clonal  material 
for  vegetatively  propagated  crops,  which  can- 
not be  stored  as  seed.  Although  their  storage  re- 
quires more  space  than  comparable  seed  stor- 


^“U.S.  Department  of  Agriculture,  Agricultural  Research  Serv- 
ice, Introduction,  Classification,  Maintenance,  Evaluation,  and  Docu- 
mentation of  Plant  Germplasm,  (ARS)  National  Research  Program 
No.  20160  (Washington,  D.C.,  U.S.  Government  Printing  Office, 
1976). 


Table  28.— Estimated  Economic  Rates  of  Return 
From  Germplasm  Accessions 


1.  A plant  introduction  of  wheat  from  Turkey  was  found  to 
have  resistance  to  ali  known  races  of  common  and 
dwarf  bunts,  resistance  to  stripe  rust  and  flag  smut, 
plus  field  resistance  to  powdery  and  snow  mold.  It  has 
contributed  to  many  commercial  varieties,  with 
estimated  annual  benefits  of  $50  million. 

2.  The  highly  successful  variety  of  short-strawed  wheat, 
‘Gaines’  has  in  its  lineage  three  plant  introductions  that 
contributed  to  the  genes  for  the  short  stature  and  for 
resistance  to  several  diseases.  During  the  3 years, 
1964-66,  about  60  percent  of  the  wheat  grown  in  the 
State  of  Washington  was  with  the  variety  ‘Gaines’.  In- 
creased production  with  this  variety  averaged  slightly 
over  13  million  bu  or  $17.5  million  per  year  in  the  3-year 
period. 

3.  Two  soybean  introductions  from  Nanking  and  China 
were  used  for  large-scale  production,  because  they  are 
well-adapted  to  a wide  range  of  soil  conditions.  All  ma- 
jor soybean  varieties  now  grown  in  t e Southern  United 
States  contain  genes  from  one  or  both  of  these  in- 
troductions. Farm  gate  value  of  soybean  crop  in  the 
South  exceeded  $2  billion  in  1974. 

4.  Two  varieties  of  white,  seedless  grapes  resulted  from 
crosses  of  two  plant  introductions.  These  varieties 
ripen  2 weeks  ahead  of  ‘Thompson  Seedless’.  Benefits 
to  the  California  grape  industry  estimated  to  be  more 
than  $5  million  annually. 


SOURCE:  U S.  Department  of  Agriculture,  Agricultural  Research  Service.  In- 
troduction, Classilicatlon,  Maintenance.  Evaluation,  and  Docu- 
mentation ol  Plant  Germplasm,  (ARS)  National  Research  Program  No 
20160  (Washington,  D C.,  U.S.  Government  Printing  0(flce,1976) 

age,  12  new  repositories  for  fruit  and  not  crops 
as  well  as  for  other  important  crops,  from  hops 
to  mint,  were  proposed  by  the  National  (ierm- 
plasm  Committee  as  additions  to  the  National 
Germplasm  System  (see  lech.  Note  12,  p.  163). 
(The  development  of  tissue  culture  storage 
methods  may  reduce  storage  costs  for  thest> 
proposed  repositories.) 

The  National  Germplasm  System  is  a \ ital  link 
in  ensuring  that  germiilasm  now  ivxisting  will 
still  be  available  in  the  futurt'.  Ilowmt'i',  the 
present  system  was  challenged  after  the  Soutli- 
ern  corn  blight  epidemic  of  1970.  Many  scien- 
tists questioned  whether  it  was  large  enough 
and  broad  enough  in  its  pi'csent  lorm  to  pio\  ide 
the  genetic  resources  that  might  he  needed. 

The  devastating  effects  of  the  corn  blight  ol 
1970  actually  led  to  the  coining  of  the  term  cro[) 
vulnerability.  During  the  e|)idemic,  as  much  as 
15  percent  of  the  entire  \ield  was  lost.  Sunn* 
fields  lost  their  whole  crop,  and  entiic  sections 
of  some  Southern  States  lost  50  pt'rcent  ol  their 


Ch.  8— The  Application  of  Genetics  to  Plants  • 157 


com.  Epidemics  like  this  one  are,  of  course,  not 
new.  In  the  19th  centurv,  the  phvllo.xera  disease 
of  grapes  almost  desti'oved  the  wine  industry  of 
France,  coffee  I'ust  disrupted  the  economy  of 
Ceylon,  and  the  potato  famine  triggered  e.xten- 
si\  e local  star\  ation  in  Ireland  and  mass  emigra- 
tion to  \orth  .Amei'ica.  In  1916,  the  red  rust  de- 
stroN'ed  2 million  hu  of  wheat  in  the  United 
States  and  an  additional  million  in  Canada.  Fur- 
ther epidemics  of  wheat  rust  occurred  in  1935 
and  1953.  The  corn  hlight  epidemic  in  the 
United  States  stimulated  a stud\’  that  led  to  the 
publication  of  a repoi't  on  the  "Genetic  \ ulner- 
ahility  of  .Major  Crops”.-'  It  contained  two  cen- 
tral findings:  that  \ ulnerahility  stems  from  ge- 
netic uniformity,  and  that  some  .American  crops 
are,  on  this  basis,  highly  \ iilnerahle.  (See  table 
29.) 

However,  genetic  variability,  is  only  a hedge 
against  \ ulnerahility.  It  does  not  guarantee  that 
an  epidemic  will  be  avoided.  In  addition,  path- 
ogens from  abroad  can  become  serious  prob- 
lems when  they  are  introduced  into  new  envi- 
ronments. .As  clearly  stated  in  the  study,  a tri- 
angular relationship  e.xists  between  host,  path- 
ogen, and  env  ironment,  and  the  coincidence  of 
their  interaction  dictates  the  severity  of  disease. 


^'.National  .Vcademv  of  Sciences.  Genetic  Vulnerabililv  of  Major 
Crops,  Washington.  D.  C.,  1972. 


The  basis  for  genetic  uniformity 

Crop  unifoi'mity  results  most  often  from  soci- 
etal decisions  on  how  to  produce  food.  The 
structure  of  agriculture  is  extremely  sensitive  to 
changes  in  the  market.  Some  of  the  basic  factors 
influencing  uniformity  are: 

• the  consumer’s  demand  for  high-quality 
produce; 

• the  food  processing  industry’s  demand  for 
harvest  uniformity; 

• the  farmer’s  demand  for  the  “best”  variety 
that  offers  high  yields  and  meets  the  needs 
of  a mechanized  farm  system;  and 

• the  increased  world  demand  for  food, 
which  is  I'elated  to  both  economic  and  pop- 
ulation grow  th. 

New'  varieties  of  crops  are  bred  all  the  time, 
but  several  can  dominate  agricultural  produc- 
tion—e.g.,  Norman  Borlaug  and  his  colleagues  in 
Mexico  pioneered  the  "green  revolution”  by 
developing  high-yielding  varieties  (HYV)  of 
wheat  that  required  less  daylight  to  mature  and 
possessed  stiffer  straw  and  shorter  stems.  Since 
the  new  varieties  (see  Tech.  Note  13,  p.  163) 
gave  excellent  yields  in  response  to  applications 
of  fertilizer,  pesticides,  and  irrigation,  the  in- 
novation was  subsequently  introduced  into 
countries  like  India  and  Pakistan.  When  a single 


Table  29.— Acreage  and  Farm  Value  of  Major  U.S.  Crops  and  Extent  to  Which 
Small  Numbers  of  Varieties  Dominate  Crop  Average  (1969  figures) 


Crop 

Acreage 

(millions) 

Value 
(millions  of 
dollars) 

Total 

varieties 

Major 

varieties 

Acreage 

(percent) 

Bean,  dry 

1.4 

143 

25 

2 

60 

Bean, snap  

0.3 

99 

70 

3 

76 

Cotton 

11.2 

1,200 

50 

3 

53 

Corns 

66.3 

5,200 

197b 

6= 

71 

Millet 

2.0 

7 

— 

3 

100 

Peanut  

1.4 

312 

15 

9 

95 

Peas 

0.4 

80 

50 

2 

96 

Potato 

1.4 

616 

82 

4 

72 

Rice 

1.8 

449 

14 

4 

65 

Sorghum 

16.8 

795 

7 

7 

7 

Soybean  

42.4 

2,500 

62 

6 

56 

Sugar  beet 

1.4 

367 

16 

2 

42 

Sweet  potato 

0.13 

63 

48 

1 

69 

Wheat 

44.3 

1,800 

269 

9 

50 

3Com  includes  seeds,  forage,  and  silage. 

^Released  public  inbreds  only. 

•^here  were  six  major  public  lines  used  in  breeding  the  major  varieties  of  corn,  so  the  actual  number  of  varieties  is  higher. 


SOURCE:  National  Academy  of  Sciences,  Genetic  Vulnerability  of  Major  Crops,  Washington,  D.C.,  1972. 


158  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


variety  dominates  the  planting  of  a crop,  there 
is  some  loss  of  genetic  variability,  the  resulting 
uniformity  causes  crop  vulnerability— and  the 
displacement  of  indigenous  varieties— a real 
problem. 

The  rate  of  adoption  of  HYVs  levels  off  below 
100  percent  in  most  countries,  mainly  because 
of  the  numerous  factors  affecting  supply  and 
demand:^^ 

• supply  factors: 

—the  present  HYVs  are  not  suitable  for  all 
soil  and  climatic  conditions; 

—they  require  seeds  and  inputs  (such  as 
fertilizers,  water,  and  pesticides)  that  are 
either  unavailable  or  not  fully  utilized  by 
every  farmer;  and 

—in  some  regions,  a strong  demand  still  ex- 
ists for  the  longer  straw  of  traditional 
varieties. 

• demand  factors: 

—consumers  may  not  prefer  the  HYVs  over 
traditional  food  varieties; 

—Government  price  policies  may  not  en- 
courage the  production  of  HYVs. 

For  these  and  other  reasons,  countries  already 
using  a great  deal  of  HYVs  will  continue  to  adopt 
them  more  slowly. 

Six  factors  affecting  adequate 
management  of  genetic  resources 

1.  Estimating  the  potential  value  of  genetic  re- 
sources is  difficult. 

Of  the  world’s  estimated  300,000  species  of 
higher  plants,  only  about  1 percent  have  been 
screened  for  their  use  in  meeting  the  diverse 
demands  for  food,  animal  feed,  fiber,  and  phar- 
maceuticals.Genetic  resources  not  yet  col- 
lected or  evaluated  are  valuable  until  proven 
otherwise,  and  the  efforts  to  conserve,  collect, 
and  evaluate  plant  resources  should  reflect  this 
assumption.  This  point  of  view  was  strongly  re- 

“D.  G.  Dalrymple,  Development  and  Spread  of  High-Yielding  Vari- 
eties of  Wheat  and  Rice  in  the  Less  Developed  Nations,  6th  ed. 
(Washington  D.C.:  U.S.  Department  of  Agriculture,  Office  of  Inter- 
national Cooperation  and  Development  in  cooperation  with  U.S. 
Agency  for  International  Development,  1978). 

Mvers,  "Conserving  Our  Global  Stock,"  Environment 
21(9):25,  1979. 


fleeted  in  a 1978  recommendation  by  the  Na- 
tional Plant  Genetic  Resources  Board.  It’s  recom- 
mendation was  that  four  major  areas  of  genetic 
storage— collection,  maintenance,  e\aluation, 
and  distribution— be  viewed  as  a "continuum 
that  sets  up  a gene  flow  from  source  to  end 
use’’.^“* 

2.  The  management  of  genetic  resources  is  com- 
plect and  costly. 

The  question  of  how  much  germplasm  to  col- 
lect is  difficult  and  strongly  influenced  by  cost. 
Thus  far,  only  a fraction  of  the  ax  ailahle  di\  ersi- 
ty  has  been  collected.  A better  scientific  undiM’- 
standing  of  the  genetic  makeup  and  pre\  ious 
breeding  history  of  major  cro|)s  will  help  deter- 
mine just  how  much  germplasm  should  he  col- 
lected. Efforts  to  gi\e  priorities  for  coIUu’tioiH* 
have  been  hindered  by  the  scientific  ga[)s  in 
knowledge  about  what  is  presently  stored 
worldwide.  And  while  attempts  ha\e  been 
made  to  estimate  the  economic  return  fi’om  in- 
troduction of  specific  plants  (see  table  28),  the 
degree  to  which  agricultui'al  production  and 
stability  are  dependent  on  generic  \ai'iahility 
has  not  been  adetjuately  analyzc'd. 

Evaluation  of  genetic  characteristics  must  he 
conducted  at  different  ecological  sit(\s  by  multi- 
disciplinary teams.  The  data  ohtaiiK'd  w ill  only 
be  useful  if  adetiuately  assessed  and  made  a\  ail- 
able  to  the  breeding  community  (see  l ech.  Note 
14,  p.  163). 

Germplasm  must  he  adec|uately  maintained  to 
assure  viability,  "woi’king  stocks"  must  he  made 
available  to  the  breeding  community.  Hu‘  |)i’i- 
mary  objective  of  storing  geriii|)Iasm  is  to  make 
the  genetic  information  axailahle  to  hreedei.s 
and  researchers. 

3.  How  much  plant  diversity  can  he  lost  without 
disrupting  the  ecological  balances  of  natural 
and  agricultural  systems  is  not  known. 


^■‘Kepoft  to  the  Sc(  rcliiry  ol  Vgrit  iillurr  b\  ihi-  \--.iNl.ini  '«•< 
retarv  lor  (:ons(*r\  alion.  Rc.-icarch,  .iiid  l.dm  .ilioti  b.i'-i-d  on  llii  di- 
liberations  and  r(>commi‘ndalions  National  I'lanI  ta-nelii  III 
sources  Boaril.  July  1978 

^“Secretarial,  International  Ho.ird  Ini'  I’lanI  (a-ncin  Kcjmc.i 
Annual  Report  197/i,  Rome,  ( onsullalne  (.roup  on  Inlcmalu'  .al 
Agricultural  Research.  I!)7i) 


Ch.8 — The  Application  of  Genetics  to  Plants  • 159 


The  arguments  parallel  those  pi-e\  iously  clis- 
cussetl  in  C\)iigress  for  protection  of  enclan- 
geretl  species  (see  lech.  Note  15,  p.  163).  The 
last  decade  has  shown  that  modes  of  [)i'oduction 
and  de\  elo[)ment  can  se\  erely  affect  the  ecolog- 

! ical  balance  of  com[)le.\  ecosystems.  \\  hat  is  not 
known  is  how  much  species  disruption  can  take 
place  before  the  ([uality  of  life  is  also  affected. 

4.  The  e.\tent  to  which  the  new  genetic  technol- 
ogies will  afYect  genetic  variability,  ^ermftlasrn 

I storage  methodoloy,ies,  ami  crop  vulnerability 
has  not  been  tietennineii. 

rhe  new  genetic  technologies  could  either  in- 
crease or  tleci'ease  crop  \ ulnerahility.  In  theoi'y, 
they  could  he  useful  in  de\eloj)ing  early  warn- 
ing systems  for  \ ulnerahility  by  screening  for 
inherent  weaknesses  in  major  crop  I'esistance. 
However,  the  relationship  between  the  genetic 
characteristics  of  plant  \ arieties  and  theii*  j)ests 
and  pathogens  is  not  understood  (see  l ech.  Note 
16,  p.  164). 

The  new  technologies  ma\  also  enhance  the 
prospects  of  using  variability,  creating  new 
sources  of  genetic  div  ersity  and  storing  genetic 
material  by: 

• increasing  v ariabilitv  during  cell  regenera- 

* tion, 

1' 

• incorporating  new  combinations  of  genetic 
information  during  cell  fusion, 

• changing  the  ploidy  lev  el  of  plants,  and 

• introducing  foreign  (nonplant)  material 
and  distantly  related  plant  material  by 
means  of  rDX.A. 

With  the  potential  benefits,  however,  come 
risks.  Because  genetic  changes  during  the  devel- 
opment of  new  varieties  are  often  cumulative, 
and  because  superior  varieties  are  often  used 
e.xtensively,  the  new  technologies  could  in- 
crease both  the  degree  of  genetic  uniformity 
and  the  rate  at  which  improved  varieties  dis- 
place indigenous  crop  types.  Furthermore,  it 
has  not  been  determined  how  overcoming  natu- 
ral breeding  barriers  by  cell  fusion  or  rDXA  will 
affect  a crop's  susceptibility  to  pests  and  dis- 
eases. 

5.  Because  pests  and  pathogens  are  constantly 
mutating,  plant  resistance  can  be  broken  down, 
requiring  the  introduction  of  new  varieties. 


Historically,  success  and  lailure  in  biXHHling 
[programs  are  linked  to  pests  and  pathogens 
overcoming  resistance.  H(mic(\  plant  breeders 
try  to  kee|)  one  step  ahead  of  mutations  or 
changes  in  |)est  and  pathogen  populations;  a 
plant  v ariety  usually  lasts  only  5 to  15  years  on 
the  market.  rher(>  is  some  ev  idence  that  patho- 
gens are  becoming  more  vii'ulent  and  aggres- 
sive— vv  hich  could  increase  the  rate  of  infection, 
enhancing  the  potential  for  an  epidemic  (see 
Tech.  Xote  17,  p.  164). 

6.  Other  economic  and  social  pressures  affect  the 
use  of  genetic  resources. 

The  Plant  \ ariety  Protection  Act  has  been 
criticized  for  being  a |)rimary  cause  of  planting 
uniform  varieties,  loss  of  germplasm,  and  con- 
glomei’ate  acxiuisition  of  seed  companies.  In  its 
op[)onents'  v iew,  such  ownership  I’ights  prov  ide 
a strong  incentive  for  seed  com[)anies  to  en- 
courage farmers  to  buy  "superior"  varieties  that 
can  he  })rotected,  instead  of  indigenous  varieties 
that  cannot,  rhe'v  also  make  plant  breeding  so 
lucrativ  e that  the  ow  nership  of  seed  companies, 
is  being  concentrated  in  multinational  corpora- 
tions—e.g.,  opponents  claim  that  79  percent  of 
the  U.S.  patents  on  beans  have  been  issued  to 
four  companies  and  that  almost  50  once-inde- 
pendent  seed  companies  have  been  acquired  by 
The  Upjohn  Co.,  ITT,  and  others.^®  One  concern 
raised  about  sucb  ownership  is  that  some  of 
these  companies  also  make  fertilizer  and 
pesticides  and  have  no  incentive  to  breed  for 
pest  resistance  or  nitrogen-fixation.  For  the 
above  reasons,  one  public  interest  group  has 
concluded^’’ 

(tlhanks  to  the  patent  laws,  the  bulk  of  the 
world's  food  supply  is  now  owned  and  devel- 
oped by  a handful  of  corporations  w'hich  alone, 
without  any  public  input,  determine  which 
strains  are  used  and  how. 

Xumerous  arguments  have  been  advanced 
against  the  above  position.  Planting  of  a single 
variety,  for  instance,  is  claimed  to  be  a function 
of  the  normal  desires  of  farmers  to  purchase 
the  best  available  seed,  especially  in  the  com- 


R.  Mooney,  Seed  of  the  Earth  (London:  International 
Coalition  for  Development  Action,  1979), 

^'Brief  for  Peoples'  Business  Commission  as  Amicus  Curiae, 
Diamondx.  Chakrabarty,  100  S.  Ct.  2204  (1980),  p.  9, 


160  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animais 


petitive  environment  in  which  they  operate. 
Moreover,  hybrid  varieties  (such  as  corn),  are 
not  covered  by  the  plant  protection  laws;  yet 
they  comprise  about  90  percent  of  the  seed 
trade. 

As  for  the  loss  of  varieties  by  vegetation 
displacement,  statutory  protection  has  been  too 
recent  to  counter  a phenomenon  that  has  oc- 
curred over  a 30-  to  40-year  period,  and  avail- 
able evidence  indicates  that  some  crops  are  ac- 
tually becoming  more  diverse.  Since  most  major 
food  crops  are  sexually  produced,  they  have 
only  been  subject  to  protection  since  1970  when 
the  Plant  Variety  Protection  Act  was  passed;  the 
first  certificates  under  that  Act  were  not  even 
issued  until  1972.  Moreover,  at  least  in  the  case 
of  wheat,  as  many  new  varieties  were  devel- 
oped in  the  7 years  after  the  passage  of  the  Plant 
Variety  Protection  Act  as  in  the  previous  17.^® 

It  is  clear  that  large  corporations  have  been 
acquiring  seed  companies.  However,  the  con- 


Kept.  No.  96-1115,  96th  Cong.,  2d  sess.,  p.  5 (June  20,  1980). 


Summary  

The  science  and  structure  of  agriculture  are 
not  static.  The  technical  and  industrial  revolu- 
tions and  the  population  explosion  have  all  con- 
tributed to  agricultural  trends  that  influence 
the  impacts  of  the  new  technologies.  Several 
factors  affect  U.S.  agriculture  in  particular; 

• To  some  degree,  the  United  States  depends 
on  germplasm  from  sources  abroad,  which 
are,  for  the  most  part,  located  in  less  devel- 
oped countries;  furthermore,  the  amount 
of  germplasm  from  these  areas  that  should 
be  collected  has  not  been  determined. 

• Genetic  diversity  in  areas  abroad  is  being 
lost.  The  pressures  of  urbanization,  in- 
dustrial development,  and  the  demands  for 
more  efficient,  more  intensive  agricultural 
production  are  forcing  the  disappearance 
of  biological  natural  resources  in  which  the 
supply  of  germplasm  is  maintained. 


nection  between  this  trend  and  the  plant  \ ariety 
protection  laws  is  disputed.  One  explanation  is 
that  the  takeovers  are  part  of  the  general  take- 
over movement  that  has  involved  all  parts  of  the 
economy  during  the  past  decade.  Since  the  pas- 
sage of  the  1970  Act,  the  number  of  seed  com- 
panies, especially  soybean,  wheat,  and  cereal 
grains,  has  increased. While  there  were  six 
companies  working  with  soybean  breeding 
prior  to  1970,  there  are  25  at  this  time.®° 

Thus,  to  date,  although  no  conclusix  e connec- 
tion has  been  demonstrated  between  the  two 
plant  protection  laws  and  the  loss  of  genetic 
diversity,  the  use  of  uniform  varieties,  or  the 
claims  of  increasing  concentration  in  the  plant 
breeding  industry;  the  question  is  still  con- 
troversial and  these  complex  problems  are  still 
unresolved. 


^“Hearing.s  on  H.R.  2844,  supra  note  35  (StatcnuMU  ol  Harold 
Loden,  Executive  Oireclor  ol  the  .American  Seed  Trade  \ssiK-ia- 
tion). 

“Brief  for  Pharmaceutical  .\Ianufaciurei-s'  Xs.soci.ition  as 
Amicus  Curiae,  Diamond  v.  Chakrabariy,  lOO  S ( t.  2204  09801.  p 
26. 


• This  lost  genetic  dix  ersity  is  irreplaceable. 

• The  world’s  major  food  ci’ops  ai’e  becoming 
more  vulnerable  as  a I’esult  of  genetic  uni- 
formity. 

The  solutions— examining  the  risks  and  exal- 
uating  the  tradeoffs— are  not  limited  to  .securing 
and  storing  varieties  of  seed  in  manmade 
repositories;  genetic  exolution- one  of  tiu*  keys 
to  genetic  diversity  and  a continuous  supplx  ol 
new  germplasm— cannot  tak(>  |)lace  on  storage 
shelves.  Until  specific  gaps  in  man's  understand- 
ing of  plant  genetics  are  filled,  and  until  tin* 
breeding  community  is  ahh*  to  identify,  collect, 
and  evaluate  sources  of  genetic  dixfisity,  it  is 
essential  that  natural  resourc(*s  prox  iding  germ- 
plasm he  preserx  ed. 


I 


Ch.  8— The  Application  of  Genetics  to  Plants  • 161 


I 

I 

Issues  and  Options — Plants 


ISSl'E:  Should  an  assessment  he  eon- 

dueti'd  to  determine  hoi%'  iiuich 
plant  ^ermplasni  mreds  to  he 
maintained? 

An  understanding  ot  how  much  germplasm 
should  he  protected  and  maintaineil  would 
make  the  management  of  genetic  resources 
simpler.  But  no  complete  answers  e.xist;  nohody 
knows  how  much  diversity  is  being  lost  by 
vegetation  displacement  in  areas  mostly  outside 
the  United  States. 

OPTIONS: 

A.  Congress  could  commission  a study  on  how 
much  genetic  variability  is  needed  or  desirable 
to  meet  present  and  future  needs. 

A comprehensive  evaluation  of  the  National 
Germplasm  System’s  needs  in  collecting,  eval- 
uating. maintaining,  and  distributing  genetic 
resources  for  plant  breeding  and  research  could 
serve  as  a baseline  for  further  assessment.  This 
ev  aluation  would  require  e.xtensiv  e cooperation 
among  the  Federal,  State,  and  private  compo- 
nents linked  to  the  National  Germplasm  System. 

B.  Congress  could  commission  a study  on  the 
I need  for  international  cooperation  to  manage 
I and  preserve  genetic  resources  both  in  natural 

ecosystems  and  in  repositories. 

This  inv  estigation  could  include  an  evaluation 
of  the  rate  at  which  genetic  diversity  is  being 
lost  from  natural  and  agricultural  systems,  and 
an  estimate  of  the  effects  this  loss  will  have.  Un- 
j til  such  information  is  at  hand.  Congress  could: 

• Instruct  the  Department  of  State  to  have  its 
delegations  to  the  United  Nations  Educa- 
tional, Scientific,  and  Cultural  Organization 
(UNESCO)  and  United  Nations  Environmen- 
tal Program  (UNEP)  encourage  efforts  to  es- 
tablish biosphere  reserves  and  other  pro- 
tected natural  areas  in  less  developed  coun- 
tries, especially  those  within  the  tropical 
latitudes.  These  reserves  would  serve  as  a 
source  for  continued  natural  mutation  and 
variation. 


• Instruct  the  Agency  for  International  De- 
velopment (AID)  to  place  high  priority  on, 
and  accelerate  its  activities  in,  assisting  less 
developed  countries  to  establish  biosphere 
reserves  and  other  protected  natural  areas, 
providing  for  their  protection,  and  support 
associate  research  and  training. 

• Instruct  the  International  Bank  for  Recon- 
struction and  Development  (World  Bank)  to 
give  high  pi'ioritv  to  providing  loans  to 
those  less  developed  countries  that  wish  to 
establish  biosphere  reserv  es  and  other  pro- 
tected natural  areas  as  well  as  to  promote 
activ  ities  related  to  biosphere  reserve  pres- 
ei'vation,  and  the  research  and  manage- 
ment of  these  areas  and  resources. 

• Make  a one-time  special  contribution  to 
LfNESCO  to  accelerate  the  establishment  of 
biosphere  reserves. 

Such  measures  for  in  situ  preservation  and 
management  are  necessary  for  long-term  main- 
tenance of  genetic  diversity.  Future  needs  are 
difficult  to  predict;  and  the  resources,  once  lost 
are  irreplaceable. 

C.  Congress  could  commission  a study  on  how  to 
develop  an  early  warning  system  to  recognize 
potential  vulnerability  of  crops. 

A followup  study  to  the  1972  National  Acad- 
emy of  Science’s  report  on  major  crop  vul- 
nerability could  be  commissioned.  Where  high 
genetic  uniformity  still  exists,  proposals  could 
be  suggested  to  overcome  it.  In  addition,  the 
avenues  by  which  private  seed  companies  could 
be  encouraged  to  increase  the  levels  of  genetic 
diversity  could  be  investigated.  The  study  could 
also  consider  to  what  extent  the  crossing  of 
natural  breeding  barriers  as  a consequence  of 
the  new  genetic  technologies  will  increase  the 
risks  of  crop  vulnerability. 

ISSUE:  What  are  the  most  appropriate 

approaches  for  overcoming  the 
various  technical  constraints 
that  limit  the  success  of  molec- 
ular genetics  for  plant  improve- 
ment? 


162  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Although  genetic  information  has  been  trans- 
ferred by  vectors  and  protoplast  fusion,  DNA 
transformations  of  commercial  value  have  not 
yet  been  performed.  Molecular  engineering  has 
been  impeded  by  the  lack  of  vectors  that  can 
transfer  novel  genetic  material  into  plants,  by 
insufficient  knowledge  about  which  genes 
would  be  useful  for  breeding  purposes,  and  by 
a lack  of  understanding  of  the  incompatibility  of 
chromosomes  from  diverse  sources.  Another 
impediment  has  been  the  lack  of  researchers 
from  a variety  of  disciplines. 

OPTIONS: 

A.  Increase  the  level  of  funding  for  plant  molec- 
ular genetics  through: 

1.  the  National  Science  Foundation  (NSF),and 

2.  the  Competitive  Grants  Program  of  the  U.S. 
Department  of  Agriculture  (USDA). 

B.  Establish  research  units  devoted  to  plant  mo- 
lecular genetics  under  the  auspices  of  the  Na- 
tional Institutes  of  Health  (NIH),  with  empha- 


Technical  notes 


1.  A recent  example  of  such  a mutation  was  the  opaque-2 
gene  in  corn,  which  was  responsible  for  increasing  the 
corn's  content  of  the  amino  acid  lysine. 

2.  There  is  disagreement  about  what  is  meant  by  produc- 
tivity and  how  it  is  measured.  Statistical  field  data  can 
be  expressed  in  various  ways— e.g.,  output  per  man- 
hour, crop  yield  per  unit  area,  or  output  per  unit  of 
total  inputs  used  in  production.  A productivity  meas- 
urement is  a relationship  among  physical  units  of  pro- 
duction. It  differs  from  measurements  of  efficiency, 
which  relate  to  economic  and  social  values. 

3.  Nevertheless,  some  parts  of  the  world  continue  to  lack 
adequate  supplied  of  food.  A recent  study  by  the  Pres- 
idential Commission  on  World  Hunger”  estimates  that 
"at  least  one  out  of  every  eight  men,  women,  and  chil- 
dren on  earth  suffers  malnutrition  severe  enough  to 
shorten  life,  stunt  physical  growth,  and  dull  mental 
ability.” 

4.  In  theory,  pure  lines  produce  only  identical  gametes, 
which  makes  them  true  breeders.  Successive  cross- 
breeding will  result  in  a mixture  of  gametes  with  vary- 
ing combinations  of  genes  at  a given  locus  on  homolog- 
ous chromosomes. 

^'Report  of  the  Presidential  Commission  on  World  Hunger.  Over- 
coming World  Hanger:  The  Challenge  Ahead,  Washington,  H.C.. 
March  1980. 


sis  on  potential  pharmaceuticals  derived  from 
plants.  I 

C.  Establish  an  institute  for  plant  molet  alar  ge-  ! 
netics  under  the  Science  and  Education  Ad- 
ministration at  USDA  that  would  include  mul- 
tidisciplinary teams  to  consider  both  basic  re- 
search questions  and  direct  applications  of  the 
technology  to  commercial  needs  and  practices. 

The  discoveries  of  molecular  jilant  genetics 
will  be  used  in  conjunction  with  traditional 
breeding  programs.  Therefore,  each  of  the 
three  options  would  recjuire  additional  ttppro- 
priations  for  agricultural  research.  K.xisting 
funding  structures  could  he  used  for  all  three, 
but  institutional  reorganization  would  he  ic- 
quired  for  options  B and  C.  The  main  argument  i 
for  increasing  fiSDA  funding  is  thtit  it  is  the  letul 
agency  for  agricultural  research,  Idi-  incretising 
NSF  and  NIH  funding,  that  they  currently  lunc 
the  greatest  expei  tise  in  molecular  techni(|ues. 
Option  C emphasizes  the  impoi  tanci'  of  the  in- 
terdisciplinary needs  of  this  I’l’seai  eh. 


5.  Normally,  chromosomes  ai'c  inheriled  m sets  I hr 
more  tnujiu'iil  diploid  stale  consists  ol  two  sets  m eai  h 
plant.  Recuiuse  chi-omosome  pairs  .ite  homologous 
(ha\  e the  same  litiear  getie  se(|ui“n(  r-l  cells  must  m.im 
tain  a degree  of  gi'uelic  integrity  hclu  een  chromosome 
pairs  during  cell  di\  ision  I hereloi  e.  iiu  re.ises  m 
ploidv  iinoh’e  entire  sets  of  chi'omosomes  diploul  (j 
sell  is  manipulated  to  Iriploid  l.'I  seil  or  e\  en  to 
tetra|)loid  (f-si'l). 

a.  The  esiimaltui  theoretical  limit  to  elliciem  v ol  phoiu 
synthesis  during  the  grow  Ih  cycle  is  ,s  7 percent  Ilou 
ever,  th(‘  I'ecord  I '.S  Stale  a\eiage  (llii  hu  .u  i e II 
linois,  197.'))  for  coi'o,  ha\  ing  a high  pholos\  nlhi-lii  i ,iie 
iti  comparison  to  other  majoi'  crops  ,ip|>i  o.u  lies  i nd\  I 
percent  efficic'ncy.^'  Since  ,i  majoi'  limilmg  siej)  m jil.mi 
productivity  lies  in  this  elliciencs  loi  the  |)holos\o 
thetic  pi'oeess,  there  is  potential  lot  |il,inl  hl■l•l■l|ln^; 
strategies  to  improve  the  elliciencv  ol  pholos\  uiliesis 
of  many  other  important  cro|)s  I his  w ould  hav c ,i  tie 
mendous  impact  on  agricultural  produciiv  ilv 
7.  It  is  difficult  to  separate  social  values  Imm  the  ei  imiuih 
ic  strucluri’s  affecting  the  produciiv  ilv  ol  \iiiei  n .m 
agriculture.  Social  pressures  and  decisions  .uc  i umpli  \ 

‘^Ollice  of  leclmnlog)  VssessmenI  I s (.ii,,  , . I , 
liiological  Prnrr.'i.sr.s,  \nliinir  II  lrihnit.il  tei  ,'.'  VV  , i.. 

I ) ( : I I.S  (,ov  emmenl  I’niitmg  ( IMm  e liiK  I'lHo 


Ch.  8— The  Application  of  Genetics  to  Plants  • 163 


and  integi  alfd— »' n . cnnlliil^  ili>\  elopinf; 

mauimim  piiHluitn  itv  aiul  en\  iit)mm*ntal  roiut*rns 
are  hv  the  ivinuval  ol  elleeti\»“  pe^tieiiles  trom 

the  market  \pplieation>  ot  existing  i»r  ne^\  ttH  hnol- 
ogies  max  1h>  sereened  h\  tin-  piihlie  loi-  aeee|>tahle 
enx  imnmental  impact  ('ontlict  also  «*\ists  lH*txxeen 
higher  pn)ductix  itx  and  higher  nutritixe  lontent  in 
loud.  sinc»*  selection  Idr  one  often  hurts  the  other 

8 \ critical  photosvnthetic  t*n/.x  iue  (rihulose  liiphosphate 
carlM>\  l.isel  is  tormeil  from  information  supplied  h\ 
different  genes  knated  iiulepeiulentlx  in  the  chloro- 
()last  la  plastidt  and  tht'  nucknis  of  the  cell  It  is  com- 
|M)seil  of  Ixxo  separate  protein  chains  that  must  link 
together  within  die  chloroplasi  Hie  larger  of  these 
chains  is  cixled  for  h\  a gene  in  the  chlomplasi— anti  it 
is  this  gene  ih.il  has  heen  rect*nlK  isolaleti  and  cloned 
The  smaller  suhunit  however  derives  from  the  plant 
nucleus  Itself  This  ctH)(H*ralion  Ix'lwtHMi  the  nucleus 
anti  the  chloroplast  to  pititluce  the  functitinal  expres- 
sion of  a gene  is  an  interesting  phenomenon  Because  it 
exists,  the  genetics  of  the  cell  coultl  he  manipulatetl  .so 
that  cv  loplasiiiicallv  inlrtHlucetl  genes  can  mfluenct' 
nuclear  gene  functions  Perhaps  mtist  iiii[M)rtantlv  at 
this  stage,  plaslitl  genes  are  prime  cantlitlales  to  clarify 
the  basic  molecular  genetic  merhanisms  in  higher 
plants 

9 rhe  ativantages  tti  using  mass  pitipagation  technit|ues 
for  straw  herrv  (ilants  ait*  that  thtise  prtitluced  frtim 
tissue  culture  are  v irus-free,  and  a (ilantlet  produced  in 
tissue  culture  ran  prixkice  more  shoots  or  runners  fur 
transplanting 

rhe  diiwidv  antages  are  that  during  the  first  vear  the 
fruit  tends  to  lie  smaller  and.  therefore,  less  comrner- 
ciallv  acceptable:  the  plants  from  tissue  culture  mav 
have  tmuhle  adapting  to  soil  conditions,  vv  hich  can  af- 
fect their  vigor,  especially  during  the  first  growing  sea- 
son: and  the  price  per  plantlet  ready  for  planting  from 
tissue  culture  systems  may  lie  more  expensive  than 
commercial  prices  for  rooted  shoots  or  runners  bought 
in  bulk. 

10.  U heat  protein  is  deficient  in  sev  eral  amino  acids,  in- 
cluding Ivsine.  Considerable  attention  has  been  de- 
voted in  the  past  5 to  10  years  to  improv  ing  the  nutri- 
tional properties  of  wheat.  Thousands  of  lines  have 
been  screened  for  high  protein,  w ith  good  success,  and 
high  Iv  sine  genes  w ith  poor  success.  Some  high  protein 
varieties  have  been  developed,  but  adoption  by  the 
farmer  has  been  mediocre  at  best,  partly  because  of 
reduced  yield  lev  els.  There  are  some  e.xceptions;— e g., 
the  \ ariety  Plainsman  \ ' has  maintained  both  high 
protein  and  yield  lev  els,  w hich  indicates  tha  there  is  no 
consistent  relationship  between  low  protein  and  high 
yields  in  some  v arieties. 

11.  Some  42  percent  of  the  total  land  area  in  the  tropics, 
consisting  of  1.9  billion  hectares,  contains  significant 
forest  cover.  It  is  difficult  to  measure  precisely  the 
amount  of  permanent  forest  cov  er  that  is  being  lost; 
however,  it  has  been  estimated  that  40  percent  of 
"closed"  forest  (hax  ing  a continuous  closed  canopy)  has 
already  been  lost,  with  1 to  2 percent  cleared  annually. 


If  the  highest  (iredictcd  rate  of  loss  continues,  half  of 
the  remaining  closetl  forest  area  vv  ill  be  lost  by  the year 
2001).’^  rhe  significance  of  this  loss  is  exfiressed  by 
\ormaii  .Myers  in  his  report.  Conversion  of  Tropical 
Moist  Forests,  |ire|iared  for  the  Committee  on  Besearch 
Priorities  in  I rupical  Biolog^v'  of  the  National  Academy 
ol  Si  ience's  .National  Besearch  Council:  "Kxtrapolation 
of  figures  from  w ell-known  groups  of  organisms  sug- 
gest that  there  are  usually  tw  ice  as  many  species  in  the 
tropics  as  teiii|)erale  regions.  If  two-thirds  of  the 
Impical  species  oci  ur  in  IMF  (tropical  moist  forests),  a 
reasonable  extrapolation  from  known  relationships, 
then  the  species  of  the  I ,\1F  should  amount  to  some  40 
to  50  percent  of  the  |)lanel's  stock  of  species— or  some- 
w here  hetw  t*en  2 million  and  5 million  species  altogeth- 
i*r  In  other  words,  nearly  half  of  all  species  on  Ivarth 
are  ap|)arentlv  containeil  in  a biome  that  comprises 
only  0 percent  of  the  globe's  land  surface.  Probably  no 
more  than  300.000  of  these  species— no  more  than  15 
percent  and  possibly  much  less— have  ever  been  given 
a l.atin  name,  and  most  are  totally  unknown. 

12  In  1975,  ihe  (iommiltee  estimaled  thal  S4  million  would 
be  necessary  for  capital  costs  of  each  repository,  with 
recurring  annual  expenses  of  $1,4  million  for  salaries 
and  operalions.  I'SD.A  has  allocaled  SI.  16  million  for  its 
share  of  the  construction  costs  for  the  first  facility  to 
be  constructed  at  the  Oregon  State  University  in  Cor- 
V allis. 

13  High  yielding  varielies  (HV\"s)  can  be  defined  as  poten- 
tially high-yielding,  usually  semidwarf  (shorter  than 
conventional),  types  that  have  been  developed  in  na- 
tional research  jirograms  worldwide.  Wheat  varieties 
were  developed  by  the  International  Maize  and  Wheat 
Improvement  Center  and  rice  varieties  by  Interna- 
tional Bice  Besearch  Institute.  Many  improved  varieties 
of  major  crops  of  conventional  height  are  not  currently 
considered  H\  \ types,  but  they  have  often  been  incor- 
porated into  H\'\  breeding.  HY\'s,  because  of  biological 
and  management  factors,  rarely  reach  their  full 
harv  est  potential. 

14.  .Although  the  National  Germplasm  System  sucessfully 
handles  some  500,000  units  to  meet  annual  germplasm 
requests,  many  accessions— like  the  35,000  to  40,000 
wheat  accessions  stored  at  the  Plant  Genetics  and 
Germplasm  Institute  at  Beltsville,  Md  — have  yet  to  be 
examined.  Furthermore,  the  varieties  released  for  sale 
by  the  seed  companies  are  not  presently  evaluated  for 
their  comparative  genetic  differences. 

15.  For  comparison,  the  National  Germplasm  System  func- 
tions on  less  than  $10  million  annually,  whereas  the  En- 
dangered Species  Program  had  a fiscal  year  1980  budg- 
et of  over  $23  million.  The  funds  allocated  to  the  En- 

^^Report  to  the  President  by  a U.S.  Interagency  Task  Force  on 
Tropical  Forests,  The  World's  Tropical  Forests:  A Policy,  Strategy, 
and  Program  for  the  United  States,  State  Department  publication 
No.  9117.  Washington,  D.C.,  May,  1980. 

.Myers,  Conversion  of  Tropical  Moist  Forests,  report  for  the 
Committee  on  Research  Priorities  in  Tropical  Biology  of  the  Na- 
tional Research  Council,  National  Academy  of  Sciences,  Washing- 
ton, D.C.,  1980. 


164  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


dangered  Species  Program  are  used  for  such  activities 
as  listing  endangered  species,  purchasing  habitats  for 
protection,  and  law  enforcement. 

16.  The  uses  of  pest-resistant  wheat  and  corn  cultivars  on 
a large  scale  for  both  diseases  and  insects  are  classic 
success  stories  of  host-  plant  resistance.  However,  re- 
cent trends  in  the  Great  Plains  Wheat  Belt  are  disturb- 
ing. The  acreage  of  Hessian  fly-resistant  wheats  in  Kan- 
sas and  Nebraska  has  decreased  from  about  66  percent 
in  1973  to  about  42  percent  in  1977.  Hessian  fly  infesta- 
tions have  increased  where  susceptible  cultivars  have 
been  planted.  In  South  Dakota  in  1978,  in  an  area  not 
normally  heavily  infested,  an  estimated  1.25  million 
acres  of  spring  wheat  were  infested  resulting  in  losses 
of  $25  million  to  $50  million.  An  even  greater  decrease 
in  resistant  wheat  acreage  is  expected  in  the  next  2 to  5 
years  as  a result  of  releases  of  cultivars  that  have  im- 
proved agronomic  traits  and  disease  resistance  but  that 
are  susceptible  to  the  Hessian  fly.  Insect  resistance  has 
not  been  a significant  component  of  commercial 
breeding  programs. 

^'Office  of  Technology  Assessment,  U.S.  Congress,  Pest  Manage- 
ment Strategies  in  Crop  Protection  (vol.  1,  Washington,  D C.;  ll.S. 
(kjvernment  Printing  Office,  October  1979),  p.  73. 


17.  Expressed  in  genetic  terms,  cases  exist  "where  the  in- 
troduction of  novel  sources  of  major  gene  resistance 
into  commercial  cultivars  of  crop  plants  has  resulted  in 
an  increase  in  their  frequency  of  corresponding  \ iru- 
lence  genes  in  the  pathogen”.^®  This  has  been  reported 
in  Australia  with  wheat  stem  rust,  barley  powdery  mil- 
dew, tomato  leaf  mold,  and  lettuce  downy  mildew.  E\  i- 
dence  suggests  that  there  is  considerable  gene  flow  in 
the  various  pathogen  populations— e.g.,  asexual  trans- 
fer can  quickly  alter  the  frequency  of  virulence  genes. 
Furthermore,  pressures  brought  about  in  the  evolu- 
tionary process  have  developed  such  a high  degree  of 
complexity  in  both  resistance  and  virulence  mech- 
anisms, that  breeding  approaches,  especially  those  only 
using  single  gene  resistance,  can  be  easily  overcome. 


3“R.  C.  Shattock,  B.  D.  Janssen,  R.  WhilInvacI,  and  D S.  Shaw. 
"An  Inlei-pretalion  of  the  Freqiieneies  of  Host  Speeifie  Phenotypes 
of  Phytophthora  infestans  in  North  Wales. " Ann.  Appli.  liiol.  86:249, 
1977, 


chapter  9 

Advances  in  Reproductive 
Biology  and  Their  Effects 
on  Animal  Improvement 


chapter  9 


Page 


Background 167 

The  Scientific  Era  in  Livestock  Production 167 

Controlled  Breeding 168 

Scientific  Production 170 

Resistance  to  Change 171 

Some  Future  Trends 172 

Technologies 173 

Technologies  That  Are  Presently  Useful 174 

Sperm  Storage 174 

Artificial  Insemination 174 

Estrus  Synchronization 176 

Superovulation 176 

Embryo  Recovery 176 

Embryo  Transfer 177 

Embryo  Storage 177 

Sex  Selection 177 

Twinning 177 

More  Speculative  Technologies 178 

In  Vitro  Fertilization 178 

Parthenogenesis 178 

Cloning 178 

Cell  Fusion 179 

Chimeras 179 

Recombinant  DNA  and  Gene  Transfer 179 

Genetics  and  Animal  Breeding 179 

The  National  Cooperative  Dairy  Herd 

Improvement  Program 180 

Other  Species 181 

Conclusion 183 

Impacts  on  Breeding 183 

Dairy  Cattle 183 

Beef  Cattle 185 


Page 


Other  Species 187 

Other  Technologies 188 

Aquaculture 189 

Poultry  Breeding 189 

Issue  and  Options  for  Agriculture— Animals  ....  190 


Tables 


Table  No.  ' Page 

30.  Heritability  Estimates  of  Some  Economically 

Important  Traits 169 

31.  Results  of  Superovulation  in  Farm  Animals  .174 

32.  Experimental  Production  of  Identical 

Offspring 178 

33.  National  Cow  Year  and  Averages  for  All 

Official  Herd  Records,  by  Breed  May  1 , 1978- 
Apr.  30,  1979 ” ^ 181 

34.  Predicted  Difference  of  Milk  Yield  of  Acti\  e 

A1  Bulls 184 

Figures 

Figure  No.  Page 

30.  Eras  in  U.S.  Beef  Production 168 

31.  Milk  Yield/Cow  and  Cow  Population,  United 

States,  1875-1975.  . . . ! 170 

32.  Milk  Production  per  Cow  in  1958-78  170 

33.  The  Way  the  Reproductive  Technologies 

Interrelate 175 

34.  Change  in  the  Potential  Numher  of  Progein 

per  Sire  From  1939  to  1979  176 


chapter  9 

Advances  in  Reproductive  Biology  and 
Their  Effects  on  Animal  Improvement 

Back^tjround  


During  the  past  30  wai's,  ii(*\\  U'('hnoIogie\s 
ha\  t'  l(ul  to  a luiulanuMital  shilt  in  the*  way  the 
I'nited  State's  produc  es  meat  and  li\estoek.  One 
sc't  ol  these'  te'('hne)le)gies— the'  suhjeet  e)f  tliis 
se'e  tieen— use's  kneew  le'dgc'  eel  the'  i’('|)re)dueti\ e 
preee'e'ss  in  larm  animals  te>  ine'reasc'  |)r{)duetion. 

I he'  impae'ts  e)t  e'xisting  hre'C'ding  teelinologies 
have'  he't'11  gi  e'at,  and  muc  h pre)gi-('ss  is  still  [)Os- 
sil)le  thre)ugh  the'ir  c'e)ntiiuu'd  use.  I'he  deve'lop- 
nu'iit  e)l  ne'w  tee  hnoleegies  is  ine'v  itahle  as  w ell. 

In  a marke't  ('e’one)my  like  that  of  the  I'nited 
States,  the  tacte)i'  that  most  inriuenees  the  adop- 
tie)n  e)f  te'e  hnole)i'\  is  eee)nomies.  .New  technolo- 
gies in  re'pi  e)ducti\  e*  physiology  w ill  he  used 
widely  onlv  il  the'v  increase  the  etTiciency  of 
breeding  programs— i.e.,  only  If  they  provide 
greater  control  over  breeding  than  present 
methods  do,  and  only  if  the  economic  advan- 
tages of  the  increased  control  can  be  recov- 
ered.* 

But  economic  factors  are  not  the  only  ones 
that  influence  technological  change— e.g.,  poul- 
try and  livestock  production  have  influenced 
and  ha\  e been  influenced  by: 

• Ciov  ernment  regulation  such  as  meat  grad- 
ing standards: 

• increased  aw  areness  of  health  effects,  such 

■ Vs  tlisciisscd  in  ;ip|).  Ill-B,  \er\  pai'ly  adopters  of  a technology' 
often  ilo  .so  foi- other  than  economic  reasons. 


as  from  the  use  of  antibiotics  in  livestock 
feed; 

• env  ironmental  concerns,  such  as  the  prob- 
lems of  w aste  removal,  especially  near  fac- 
tory fai’ms: 

• the  growth  of  knowledge,  in— e.g.,  the  re- 
productive processes  of  farm  animals  and 
the  accuracy  of  evaluating  the  genetic 
merit  of  breeding  animals;  and 

• complementai'v  technologies  such  as  re- 
frigerated storage  and  transportation. 

i\ew  technologies,  from  breeding  to  food  de- 
livery systems,  have  reshaped  the  traditional 
.American  farm  into  a modern  production  sys- 
tem that  is  increasingly  specialized,  capitalized, 
and  integrated  with  off-farm  services.  Applied 
genetics  in  animal  production  has  been  one  of 
the  forces  behind  these  changes.  The  technolo- 
gies that  have  sprung  from  it  include  not  only 
the  new,  esoteric  techniques  for  cellular  manip- 
ulation discussed  in  other  parts  of  this  report, 
but  also  more  well-known  technologies,  like  ar- 
tificial insemination.* 

• Technologies  selected  for  discussion  in  this  part  of  the  report 
in\'ohe  direct  manipulation  of  sex  cells.  More  speculative  technol- 
ogies for  manipulations  at  the  subcellular  level  are  assessed  here 
as  well.  ,\o  effort  was  made  to  cover  all  technologies  with  potential 
for  improving  the  genetic  qualities  of  livestock— e.g,,  management 
techniques  like  estrus  detection  and  pregnancy  diagnosis  were 
omitted,  as  were  various  other  methods  for  improving  reproduc- 
tion efficienev. 


The  scientific  era  in  livestock  production 


Producing  purebred  beef  livestock  has  been 
the  dominant  breeding  objective  throughout 
most  of  the  20th  century.  The  open  range  of  the 
.American  \\  est  and  Southwest— the  "romantic” 
era  in  beef  cattle  production— lasted  until  about 
1890.  (See  figure  30.)  Then  the  range  was 


fenced-in  and  the  longhorn  was  replaced  with 
new  breeds  by  the  turn  of  the  century— the  be- 
ginning of  the  "purebred”  era. 

Pedigree  records  and  visual  comparison  of 
conformation  to  breed  type  were  the  basic  tools 


167 


168  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


Figure  30.— Eras  in  U.S.  Beef  Production 


A 

Time 


1950 

‘Scientific’ 


Era  of  artificiai 
insemination,  population 
genetics,  and  progeny 
testing. 

1890-1950 

“Purebred” 


The  era  of  the  breed 
associations 


1860-1890 

"Empire” 


The  “romantic”  era  of 
the  cowboy  and  the 
Texas  longhorn. 


SOURCE:  Adapted  from  R.  L.  Willham,  "Genetic  Activity  in  the  U.S.  Beef  Indus- 
try," journal  paper  No.  J-7923  of  the  Iowa  Agriculture  and  Home  Eco- 
nomics Experiment  Station,  Ames,  Iowa,  Project  No.  2000,  n.d.  See 
also  Yao-chi  Lu  and  Leroy  Quance,  Agriculture  Productivity:  Expand- 
ing the  Limits,  USDA,  ESCS,  Agriculture  Information  Bulletin  431. 

of  breeding  programs.  They  were  reinforced  by 
an  institutional  system  of  breed  associations, 
and  yearly  competitions  at  county  fairs  and 
stock  shows,  and  by  import  regulations  and  |)ro- 
bibitions  against  artificial  insemination  (Al)  that 
I'estricted  innoxation.  In  rearing  animals  for 
sale  to  the  slaughterhouse,  eai'ly  breeders  and 
fai’mers  more  often  than  not  were  satisfied  with 
|)i'oducing  a calf  or  pig  that  survixed,  xxeaned 
early,  and  grew  rapidly.  Because  of  the  high 
rate  of  nexvborn  deaths,  the  production  of  an 
"axerage”  animal  xvas  a considei'able  acbiexe- 
ment  in  its  oxvn  right;  the  intricacies  of  sophisti- 
cated breeding  methods  xxere  beyond  the  ca- 
pacity of  small  operations  and  xxere  difficult  to 
carry  out  on  large  spreads.  Producing  a prize- 
xvinning  pui'ehred  xvas  left  to  the  farmer  xxith 
the  time,  money,  or  luck  to  breed  animals  that 
met  the  strict  standards  of  the  breed  associ- 
ations and  the  trained  eyes  of  the  judges  at  stock 
shows. 


During  the  fii'st  half  of  the  20th  centui'v, 
breeding  objectives  became  more  comple.x; 
farmers  and  hi'eeders  began  to  look  at  c|ualities 
other  than  mere  e.xternal  physical  attributes. 
Breeding  for  multi|)le-purposes  led  directly  to 
the  beginning  of  the  “scientific"  era  in  breeding. 

'Hie  increased  use  of  Al  for  dairy  ('attl(\ 


xvhich  took  place  about  BO  years  ago— the  he*gin- 
ning  of  the  scientific  era— xxas  an  uncertain 
start  for  applied  genetics  in  animal  hr(>eding. 
While  practitioners  and  purchasers  of  Al  xxere 
(|Liick  to  grasp  its  promise  of  imnu'diate  bene- 
fits, and  xx’hile  using  Al  xx  as  cheaper  than  oxx  n- 
ing  a hull,  its  expected  genetic  effects  xxere  not 
realized  immediately.  Dairx'men  had  assumed 
that  semen  Irom  hulls  sek'cted  from  th(‘  best 
herds  and  chosen  on  th(’  basis  of  ancestral  per- 
foi'iiiance  xvould  result  in  rapid  genetic  im- 
proxement.  They  xxeie  xxrong;  progr(>ss  xxas 
much  less  than  projected.  Because  milk  produc- 
tion is  a sex-limited  ti'ait,  r('('ords  on  f('male 
relatixes  xxere  needed  for  the  exaluation  of 
sires,  linfortunately,  th(’  records  on  I’clatixc’s 
xx  ere  usually  limited  to  comparisons  xx  ithin  oiu* 
herd,  xxei'('  confounded  by  manageiiK'iit  and 
other  enx  ironmental  factors,  and  xx(m-(>  \xcak- 
ened  by  small  sample  sizes.  I h('  major  factor 
|•esponsihle  for  th(’  diffcrcncj'  lu'txxc'cn  top-  and 
iiK'diocrc'-performing  lu'rds  turned  out  to  he 
managcMiu'iit , not  gc'iictics:  separating  the  ef- 
fects of  genetics  from  the  effects  of  genei’allx 
improx  (>d  husbandry  \x  as  extrc'inely  difficult 

Controlled  breeding 

rh(>  ohj('ctix  ('  of  any  hia'cding  program  i.s  to 
inci’eas(>  produc'tion.  I he  scientilic  er;i  h;is  jiro- 
X ided  the  hre('der  \x  ith  a x iirietx  of  nexx  ti'chnol- 
ogies  that  help  in  manipulating  ;ind  controlling 
the  repioductix  ('  pi’ocess(>s  of  the  animals  to  in- 
('rease  genetic  gain.  I he  hrec'der’s  basic  tool  is 
selection,  or  deciding  xvhich  animals  to  mate— 
e.g.,  in  beef  cattle,  a breeder  can  noxx-  selec't  for 
a wide  variety  of  [lerformance  or  (>conomic 
traits.  (See  table  30.)  Howex  er,  simply  breeding 
better  beef  cattle”  is  not  a xvorkahle  object ixc 
from  a manager’s  point  of  xiew.  render  meat, 
lean  steaks  and  roasts,  high  fer  tility,  or'  heaxy 
xveight  at  xveaning  ar'e  all  specific,  rneasur  ahlr* 
objectives  of  breeding.'  ^ OthrM'  goals,  sucli  as 
those  pertaining  to  tempcMarnent,  di.sease 
resistance,  food  efficiency,  and  car'cass  (jualitx , 

■ r'.  C.  CarUvi'if'tit.  "Scli-clion  Ci  iIim  ui  liir  H<'cl  ( alllr  lor  Itn'  I ii 
lure,"  Journal  of  Animai  Srirnce  ;rO:ril(i.  I !)70 

^r.arrv  X'.  CundilT  and  Kcitli  I-.  (ircj^oi-y  HrrI'  ( alllr  lirmtinfi. 
USDA,  Agriculture  Infornialion  llulliAin  No  2HI>  re\iM'd  St- 
vemtier  1977. 


I 


Ch.  9 — Advances  in  Reproductive  Biology  and  Their  Effects  on  Animal  Improvement  • 169 


Table  30.— Heritability  Estimates  of  Some 
Economically  Important  Traits 


Trait  Heritability 

Calving  interval  (fertility) 10% 

Birth  weight 40 

Weaning  weight 30 

Cow  maternal  ability 40 

Feedlot  gain 45 

Pasture  gain 30 

Efficiency  of  gain 40 

Final  feedlot  weight 60 

Conformation  score: 

Weaning 25 

Slaughter 40 

Carcass  traits: 

Carcass  grade 40 

Ribeyearea 70 

Tenderness 60 

Fat  thickness 45 

Retail  product  (percent) 30 

Retail  product  (pounds) 65 

Susceptibility  to  cancer  eye 30 


SOURCE.  Larry  V CundiM  and  Keiin  E.  Gregory,  Beet  Cattle  Breeding.  USOA. 

Agriculture  Information  Bulletin  No  286.  revised  November  t977,  p 9 

may  also  have  economic  value,*  but  they  are 
much  harder  to  measure. 

The  e.xtent  to  which  important  economic  or 
performance  traits  are  genetically  determined 
and  heritable  \ aries  from  trait  to  trait  and  from 
animal  to  animal.  (See  table  30.)  Heritability  is 
defined  as  the  percentage  of  the  difference 
among  animals  in  performance  traits  passed 
from  parent  to  offspring*— e.g.,  bulls  and 
heifers  with  superior  weight  at  weaning  might 
average  5 pounds  (lb)  more  than  their  herd- 
mates.  Because  weaning  weight  has  an  average 
heritability  estimate  of  30  percent,  the  offspring 
of  these  top  performing  animals  can  be  ex- 
pected to  average  1.5  lb  heavier  at  weaning  than 
their  contemporaries  (0.30  x 5 = 1.5).  This 
improvement  can  normally  be  expected  to  be 
permanent  and  cumulative  as  it  is  passed  on  to 
the  next  generation.  The  improvement  accumu- 
lates like  compound  interest  in  a savings  ac- 
count; gains  made  in  each  generation  are  com- 
pounded on  the  gains  of  previous  generations. 


^Michael  I.  Lerner  and  H.  P.  Donald,  \todern  Developments  in 
Animal  Breeding  (.\eu  York;  .Vcademic  Press.  1966). 

'Heritability  and  genetic  association  are  important  in  decisions 
about  individual  matings.  Most  breeding  programs  are  concerned 
with  spreading  genetic  gain  rapidly  throughout  a population 
(herd,  flock):  thus  two  other  refinements  for  selection  enter  the 
picture — generation  inter\  al.  and  selection  differential. 


hike  laud,  e(|uipment,  and  cash,  breeding 
stock  represents  capital  available  to  the  com- 
mercial farmer.  Bt'cause  all  in|)uts  must  be  used 
efficiently,  modern  herd  or  flock  managers  can- 
not afford  to  leave  reiiroduction  to  chance 
mating  in  the  pen  or  on  the  range.  These  pres- 
sures  for  efficient  production  have  been  de- 
scribed as  follow  s:-* 

\\  here  dairymen  are  judged  by  the  luimher  of 
cows  milked  in  an  houi'.  there  is  no  place  for  the 
slow  milking  cow  or  the  man  who  will  patiently 
milk  her  out.  T here  is  no  place  for  the  time-con- 
suming hurdle  flock  of  shee[),  for  the  small  flock 
of  chickens  maintained  under  e.xtensive  condi- 
tions, or  for  the  sow  that  must  he  watched 
while  she  farrows.  By  degrees  all  classes  of 
stock  are  being  subjected  to  .selection  w'hich 
favors  animals  that  need  a minimum  of  individ- 
ual attention. 

T he  scientific  basis  for  modern  breeding  has 
dev  eloped  slow  ly  over  the  last  century.  Applied 
genetics— one  jiart  of  today’s  programs— has 
helped  modernize  livestock  and  poultry  breed- 
ing bv  elaborating  on  the  variation  of  continu- 
ously distributed  traits  in  a population;  carrying 
over  vv  hat  was  known  about  rapidly  reproduc- 
ing laboratory  species,  like  fruit  flies  or  mice,  to 
the  much  slower  reproduction  of  large  farm 
animals;  and  developing  the  statistical  tech- 
niques for  predicting  breeding  values  or  merit 
and  analyzing  breeding  programs.® 

Two  examples  show  the  powder  of  breeding 
tools  and  the  increased  efficiency  and  produc- 
tivity of  today’s  breeders’  stocks. 

• Over  the  past  30  years,  the  average  milk 
yield  of  cows  in  the  United  States  has  more 
than  doubled.  At  the  same  time,  the  num- 
ber of  dairy  cows  in  the  United  States  has 
been  reduced  by  more  than  50  percent. 
(See  figure  31.)  Of  this  increase  in  output 
and  efficiency,  more  than  one-fourth  can 
be  attributed  to  permanent  genetic  change 
for  at  least  one  breed  (Holsteins)  partici- 
pating in  the  Dairy  Herd  Improvement  Pro- 
gram. (See  figure  32.) 

• Poultry  production  in  the  United  States  has 
become  the  most  intensive  industry  among 

■•Ibid.,  p.  20. 

=Ibid..  p.  126. 


Change  in  milk  production  (lb) 


170  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Figure  31.— Milk  Yield/Cow  and  Cow  Population, 
United  States,  1875-1975 


Year 


SOURCE:  J.  T,  Reid,  "Progress  in  Dairy  Cattle  Production,”  Agricultural  and 
Food  Chemistry:  Past.  Present,  and  Future.  R,  Teranishi  (ed,) 
(Westport,  Conn.:  Avi  Press,  1978). 


Other  species  of  poultry  as  well,  production 
processes  have  become  equally  efficient. 
As  A.  W.  Nordskog  has  noted: 

Compared  with  the  breeding  of  other  eco- 
nomically important  animals,  poultry  breed- 
ing has  been  the  first  to  leave  the  farm  ...  to 
become  part  of  a sophisticated  breeding  in- 
dustry. On  a commercial  level,  chickens  ha\  e 
been  the  first  to  be  commercially  exploited 
by  the  application  of  inbreeding-hybridiza- 
tion techniques,  as  earlier  used  in  corn,  as 
well  as  by  methods  of  selective  improvement 
using  the  principles  of  quantitative  genetics. 
Thus,  the  poultry  industry,  compared  to 
other  animal  industries,  seems  to  have  been 
the  quickest  to  apply  modern  methods  of 
genetic  improvement,  including  the  employ- 
ment of  formally  trained  geneticists  to  handle 
breeding  technology  plus  the  use  of  com- 
puters and  other  modern  business  methods.® 


Figure  32.— Milk  Production  per  Cow  (Holsteins)  in  j • 

1958-78  (New  York  and  New  England)  Scientific  production 


2-year  old  Holstein  cows  in  DHIA  by  A. I.  Sires 

-1-4000 


-1-3000 


-1-2000 
-1- 1000 
Base 

1958  1962  1966  1970  1974  1978 

Year 

SOURCE:  R.  H.  Eoote,  Department  of  Animal  Science,  Cornell  University, 
Ithaca,  N.Y.  from  unpublished  data  of  R.  W.  Everett,  Cornell 
University. 

those  for  farm  species.  For  turkeys,  the  use 
of  A1  in  hreeding  for  hreast  meat  has  been 
so  successful  that  commercial  turkeys  can 
no  longer  breed  naturally.  The  big- 
breasted male,  even  when  inclined  to  do  so, 
finds  it  physically  impossible  to  mount  the 
female.  As  a result,  a full  100  percent  ot  the 
commercial  turkey  flock  in  the  United 
States  is  replaced  each  year  using  Al.  In 


Farm  resources  incliuk'  land,  labor,  capital, 
and,  increasingly,  n(>\\  know  l(Hlge.  I'oday,  those 
who  innovate  recapturi'  tlu'  costs  of  innovating 
by  maintaining  output  vvhiU'  lowering  costs  or 
by  inci’easing  output  vv  bile  bolding  costs  (low  n. 
Some  results  of  the  drive  tow  ard  elTiciencv  have 
included  increasing  spec'ialization,  intensified 
use  of  capital  and  land  relative  to  labor,  and  in- 
tegration of  production  phases. 

Foultry  and  liv  (‘stock  operations  have  slow  Iv 
b('come  sp(‘cializ('d  ov  (‘r  the  past  .">()  years.  I be 
farmer  who  used  to  do  bis  ow  n br(‘eding,  rais- 
ing, feeding,  and  slaughtering  is  disa|)pearing. 
Now,  the  b(‘(*f  cattle  industry  in  tlu*  United 
States  consists  of:  the  pur(*bred  breeder  who 
provides  br(‘eding  stock,  the  commercial  pro- 
ducei’,  tb(‘  fe('d(’r,  tlu'  packer,  and  the  retailer 
Similar  sp(*cialization  has  occurred  lor  most 
other  species— e.g.,  less  than  l.b  primary  hi-e(‘d- 
ers  maintain  the  breeding  stock  that  produces 
the  3.7  billion  chickens  consumed  ea('b  year  in 
the  United  States.  Fbe  emergenc(*  of  other  s[)e- 
cialized  services— such  as  AI  prov  iders,  manage- 

“A.  vv.  Nordskog,  "Success  ;iiul  l ailurc  fit  (^u.inin.iliv c (.cnclic 
Theory  in  I’oultry"  in  /’rocccd/Vig.s  of  I hr  Inlrnuilinnnl  ( ontrrrnir 
on  Quantitative  Genetics,  Kdward  I’ollacki’l  el  al  led  I I Vmei  • 
Iowa:  Iowa  Slale  Universily  1‘iess,  l!)77l,  |i|i  J7-.'il 


c/7.  9 — Advances  in  Reproductive  Biology  and  Their  Effects  on  Animal  Improvement  • 171 


ment  consultants,  equipment  manufacturers— 
has  accelerated  the  trend  toward  specialization, 
and  has  given  the  commercial  operator  more 
time  to  concentrate  on  his  specific  contribution 
to  the  chain  of  production. 

Intensification  is  the  increasing  use  of  some 
inputs  to  production  in  comparison  to  others. 
Increasing  the  use  of  land  and  capital  relati\  e to 
labor  describes  the  dex  elopment  of  LfS.  agricul- 
ture, including  li\  estock  raising,  in  this  century. 
The  'factory"  farm  typifies  this  trend.  Herds 
and  flocks  are  l)red,  horn,  and  raised  in  en- 
closed areas,  ne\er  seeing  a barnyard  oi'  the 
open  range.  The  best  e.xamples  of  land-  and  ca[)- 
ital-intensi\e  systems  are  those  of  poultry 
(layers,  broilers,  and  turkeys),  confined  hog  pro- 
duction, drylot  dairy  farming,  and  some  \eal 
production. 

The  greater  use  of  land  has  been  encouraged 
by  several  factors,  including  impro\  ed  corn  pro- 
duction for  confined  hog  feeding,  programs  of 
pre\entive  medicine  curtailing  the  spread  of 
diseases  in  close  spaces,  and  en\  ironmental  con- 
trol (light,  temperature,  water,  humidity)  to  in- 
crease output  under  closely  controlled  condi- 
tions. However,  extensive  ranching  for  beef  and 
sheep  is  still  common  in  the  United  States;  the 
difficulties  associated  with  detecting  estrus 
("heat")  in  these  species  and  their  relati\  ely  slow 
rates  of  reproduction  ha\  e made  it  uneconom- 
ical to  in\  est  in  them  the  capital  necessary  for 
intensi\  e farming.  Furthermore,  beef  and  sheep 
on  extensi\e  systems  forage  on  marginal  land 
that  might  otherwise  hav  e no  use.  Beeflot  feed- 
ing, or  the  fattening  of  cattle  before  slaughter  at 
a centralized  location,  is  the  only  aspect  of  the 
beef  industry  that  is  land-intensive;  in  1977,  ap- 
proximately one-fourth  of  U.S.  beef  cattle  were 
"fed.”' 

Linking  phases  of  production  to  eliminate 
waste  or  inefficiencies  in  the  system  has  pro- 
gressed with  great  speed.  For  some  species, 
such  linkages  now  extend  from  breeding  to  the 
supermarket  (and,  in  the  case  of  fast  food 
chains,  to  the  dinner  table).  Integration  includes 


"Lyle  P.  Schertz,  et  al..  Another  Revolution  in  U.S.  Farming? 
USDA,  ESCS,  .Agricultural  Economic  Report  No.  441,  December 
1979. 


the  linking  of  supiily  industries  (feeds,  medi- 
cines, breeding  stock)  with  production  and  then 
with  marketing  services  (slaughtering,  dressing, 
packaging).  Entire  industries  and  the  Govern- 
ment in  combination  have  produced  a complex 
chain  of  operations  that  makes  use  of  Govern- 
ment inspectors,  the  pharmaceutical  industry, 
equipment  manufacturers,  the  transportation 
industry,  and  the  processed  feed  industry  in  ad- 
dition to  the  traditional  commercial  farmer. 

Because  of  this  complex  linkage,  meat  grades, 
cuts,  and  packaging  have  become  fairly  stand- 
ard in  the  .American  supermarket.  Shoppers 
have  come  to  expect  these  standards;  consum- 
ers wanting  special  services  have  learned  to  pay 
more  for  them.  Thus,  the  American  farm  has 
changed  radically  ov  er  the  past  30  years.  This 
change  has  been  described  as  follows:® 

As  farming  enterprises  grow  larger,  their 
management  have  to  equip  themselves  with  in- 
formation and  resort  to  technologists  to  help 
them  reach  decisions  and  plan  for  more  distant 
goals.  Industrial  developments  of  this  kind 
widen  the  range  of  farming  activities,  since  the 
old  style  farmer,  sensitiv'e  to  local  markets  and 
operating  on  hunches,  remains  as  a contrast  to 
those  for  whom  farming  is  rapidly  becoming 
more  of  a programme  than  a way  of  life. 

Resistance  to  change 

New  technologies  in  U.S.  agriculture  and  new 
ways  of  producing  food  and  fiber  have  been 
both  a cause  and  an  effect  of  the  movement 
from  farms  to  cities  in  the  20th  century.  Com- 
mercial farmers,  operating  on  thin  or  nonexist- 
ent profits  and  under  extreme  competition, 
have  had  strong  reason  to  innovate.  They  have 
been  forced  by  the  availability  of  new  technol- 
ogies either  to  do  so  or  to  watch  their  potential 
earnings  go  to  the  neighboring  farmer.  Various 
policies  that  have  been  adopted  to  soften  the  im- 
pacts of  the  "technological  treadmill,”  have 
somewhat  slowed  the  exodus  from  the  farms. 
They  may  have  been  adopted  for  social  reasons, 
but  they  have  also  become  increasingly  costly  to 
society.  The  taxpayer  pays  for  them;  the  con- 
sumer pays  as  well  for  every  failure  to  innovate 
on  the  farms. 

*Cundiff,  et  at,  op.  cit.,  p.  9. 


172  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Besides  a lack  of  capital  or  a lack  of  interest  in 
innovating,  some  farmers  have  resisted  applied 
genetics  because  efficiency  is  not  their  most  im- 
portant priority.  This  attitude  has  been  de- 
scribed as  follows:^ 

It  is  easy  to  see  why  breeders  are  unreceptive 
to  the  science  of  genetics.  The  business  of 
breeding  pedigree  stock  for  sale  is  not  just  a 
matter  of  heredity,  perhaps  not  even  predomi- 
nantly so.  The  devoted  grooming,  feeding  and 
fitting,  the  propaganda  about  pedigrees  and 
wins  at  fairs  and  shows,  the  dramatics  of  the 
auction  ring,  the  trivialities  of  breed  characters, 
and  the  good  company  of  fellow  breeders,  con- 
stitute a vocation,  not  a genetic  enterprise. 

Farmers  are  traditionally  an  independent 
group.  Many  believe  that  they  may  not  directly 
recapture  the  benefits  of  participating  in  a 
breeding  program  based  on  genetics;  having  no 
records  on  one’s  animals  is  often  preferable  to 
discovering  proof  that  one's  herd  is  performing 
poorly.  On  the  other  hand,  one  impact  of  AI  has 
been  to  demonstrate  to  farmers  the  value  of 
adopting  new  technologies.  Furthermore,  the 
economic  reward  of  production  records  has  in- 
creased, since  AI  organizations  purchase  only 
dairy  sires  with  extensive  records  on  relatives. 

Some  future  trends 

Applied  genetics  in  poultry  and  livestock 
breeding  comprise  a group  of  powerful  technol- 
ogies that  have  already  strongly  influenced 
prices  and  profits.  Nevertheless,  the  effect  of 
genetics  is  only  just  beginning  to  be  felt;  much 
improvement  remains  to  be  made  in  all  species. 
It  has  been  observed  that  modern  genetics: 

. . . provides  a verifiable  starting  point  for  the 
development  of  the  complex  breeding  operation 
that  many  populations  now  require  . . . (which) 
are  as  far  removed  from  simple  selection  as  the 
motor  car  is  from  the  bicycle. 

Of  these  technologies,  some  are  already  in 
regular  use,  some  are  in  the  process  of  being  ap- 
plied, and  others  must  await  further  research 
and  development  before  they  become  generally 
available. 

'•Ibid.,  p.  170. 

'“E.  P.  Cunningham,  "Current  Developments  in  the  Genetics  of 
Livestock  Improvement,"  in  tSth  Inlernational  Conference  on  Ani- 
mal Blood  Groups  and  Biochemistry,  Genetics  7:191,  1 976. 


Societal  pressures  are  one  of  the  many  fac- 
tors that  influence  the  introduction  of  these 
technologies.  Several  developments  around  the 
world  will  have  a clear  impact  on  inno\  ation  in 
general  and  on  genetics  in  particular: 

• An  expanding  population,  with  its  growing 
demand  for  food  products  of  all  kinds. 

• The  growth  in  income  for  parts  of  the  pop- 
ulation, which  may  increase  the  demand 
for  sources  of  meat  protein. 

• Increasing  comjietition  for  the  consumer's 
dollar  among  various  sources  of  protein, 
which  could  reduce  demand  for  meat. 

• Increasing  competition  for  prime  agri- 
cultural land  among  agricultural,  urban, 
and  industrial  interests.  Fess-lhan-prime 
land  may  also  he  brought  hack  into  prodiu'- 
tion  as  demand  rises,  and  lh(*  sanu’  pres- 
sures may  cause  land  pricers  to  rise  high 
enough  to  encourage  greater,  or  intensi- 
fied, use  of  land  in  li\'(fslock  pi'oduction. 

• Increasing  demand  for  I'.S.  food  and  fiber 
products  from  abroad,  U'ading  to  oppor- 
tunities for  incr(?as(ui  |)rofits  for  successful 
producers. 

Changes  like  thes(‘  will  strongly  allect  the 
way  American  fai  niers  |)roduce  food  and  fihei- 
products,  rhe  economics  of  efficiency  and  a 
growing  world  population  w ill  continue  to  place 
pressure  on  tlu?  agricultural  sector  to  inno\ate 
In  animals  and  animal  |)roducts,  efficiencies  w ill 
he  found  in  all  stc'ps  of  production.  I.fforts  will 
be  made  to  incr(?ase  the  numhei-  of  li\(*  births 
and  to  iXHluce  neonatal  calf  fertility,  presently 
one  of  th(?  costliest  steps— in  terms  ol  animals 
lost— throughout  tlu?  world.  I^stimates  ol  the  po- 
tential moncUary  h(Miefits  of  the  application  of 
knowledge  obtained  fi'om  |)rior  reseaich  in  re- 
producti\’(^  physiology  l ange  as  high  .is  ,S  i bil- 
lion per  yt^ar.  Another  area  for  great  economies 
in  production  is  genetic  gain.  Much  ^enetK 
progi’ess  remains  to  In'  made  in  all  species 

t'ertain  t(U’hnologies  promise  to  incre.ise  the 
ability  of  farmers  to  capitali/.e  on  the  genetic  im 
pro\'(Miient  of  economically  im|)ort.mt  Ir.iits 
Suppliers  of  gc'iietic  material  (semen,  emhi  vus) 
will  focus  increased  attention  on  the  \ .ilue  ut 
their  products  for  sale  both  in  the  I nited  St.ites 
and  abroad. 


Ch.  9— Advances  in  Reproductive  Biology  and  Their  Effects  on  Animal  Improvement  • 1 73 


I lie  dt'\  aiul  applii  ation  ol  ('(M  tain 

kt‘\  lfrhiu)l()f'ies  will  alteft  rt'laloil  tei'hnol- 
ogies— e.g.,  the  a\  ailahiliU’  of  reliable  estrus 
detection  and  estrus  synchronization  methods 
should  inci'ease  the  use  of  Al  and  emhr\o  trans- 
fer in  beef  anti  tlair\  cattle,  thereby  spreatling 
genetic  advantage.  Further  pi'ogress  in  the 
freezing  of  eml)ryos  sliould  facilitate  the  genetic 
ev  aluation  of  cows  and  heifers. 

Other  ti'ends  that  mav  influence  U>chno- 
logical  change  include  the  shifting  av  ailahilitv  of 


I'eseai'ch  funds,  changing  consumer  tastes,  and 
gi’ovv  th  of  rt'gulations  (for  instance,  stricter  con- 
trols on  environmental  (|uality  or  hormonal 
treatments).  Th(‘  e.xpansion  of  an  animal  rights 
movement  may  influence  the  degree  to  vvliich 
confinement  housing,  and  therefore  controlled 
hret'ding,  is  acceptable.  .And  increased  energy 
costs  may  (‘ithei-  encoui'age  development  of  the 
technologies  (through  efforts  for  greater  effi- 
citMicv ) or  discourage  them  (through  greater  use 
of  foi'age  and  e.xtensive  systems). 


Technologies  

Sexual  reproduction  is  a game  of  chance.  Be- 
cause s[)erm  and  ova  each  contain  only  a ran- 
dom half  of  the  gtMies  of  each  paiaMit.  tlie  num- 
ber of  |)ossihle  combinations  that  can  result  is 
nearlv  infinite.  Some  pi-og(MU  aiv  likelv  to  sur- 
vive and  reproduce:  others  die  either  before 
birth  or  vv  ithout  ()roducing  offs|)ring. 

rhe  great  variation  achieved  through  sexual 
reproduction  produces  certain  animals  that 
satisfy  the  needs  and  desires  of  the  breeder  far 
more  than  others.  On  the  other  hand,  the  off- 
spring of  these  outstanding  animals  are  usually 
less  so  than  their  parents,  although  they  are 
generally  still  ahov  e av  erage. 

Animal  breeders  hav  e inv  ested  great  effort  in 
improv  ing  succeeding  generations  of  domestic 
animals,  both  by  limiting  the  differences  due  to 
the  chance  associated  vv  ith  sexual  reproduction 
and  by  taking  adv  antage  of  the  favorable  combi- 
nations that  occur.  E.xamples  of  these  efforts  in- 
clude keeping  records,  establishing  progeny 
testing  schemes,  amplifying  the  reproduction  of 
outstanding  indiv  iduals  by  .A I and  embryo  trans- 
fer, and  establishing  inbred  lines  to  capitalize  on 
their  more  reliable  ability  to  transmit  charac- 
teristics to  their  offspring. 

Because  of  these  efforts,  and  because  dairy 
cattle  breeders  hav  e adopted  innovativ  e tech- 
nologies through  the  vears,  far  more  is  known 
about  reproduction  in  the  cow  than  in  other 
farm  animals.  The  demand  for  milk  and  beef 
has  provided  an  impetus  for  the  speedy  intro- 


duction of  technologies  that  might  prove  eco- 
nomically adv  antageous. 

Several  observations  can  be  made  about  the 
state  of  the  art  for  1(S  technologies  that  enhance 
the  inherited  ti’aits  of  animals.  (See  also  app. 
Il-C.) 

The  technologies  are  at  different  stages  of  re- 
search and  development. 

The  practice  of  ,AI  in  dairy  cattle  has  had  the 
greatest  practical  impact  of  all  the  genetic  tech- 
nologies used  in  the  breeding  of  mammals.  In 
contrast,  not  a single  farm  animal  has  been  suc- 
cessfully raised  after  a combination  of  in  vitro 
fertilization  and  embryo  transplant.  The  use- 
fulness of  several  of  the  technologies  for  animal 
production,  such  as  recombinant  DNA  (rDNA) 
and  nuclear  transplantation,  is  purely  specu- 
lative at  this  writing. 

The  usefulness  of  the  technologies  differs  from 
species  to  species. 

These  differences  can  often  be  explained  by 
biological  factors— e.g.,  sperm  storage  capabil- 
ities are  currently  limited  for  swine  because 
freezing  kills  so  many  of  the  sperm.  Manage- 
ment techniques  are  important  as  well;  exten- 
sive beef-raising  systems  have  in  the  past  made 
estrus  detection  and  synchronization  imprac- 
tical, thereby  limiting  the  use  of  AI.  (Fewer  than 
5 percent  of  the  U.S.  beef  herd  are  artificially  in- 
seminated, compared  with  60  percent  of  the  na- 


174  • Impacts  of  Applied  Genetics — Micro-Organisms,  Piants,  and  Animais 


tional  dairy  herd.)  And  economics  can  also  play 
a role;  in  general,  the  lower  an  animal’s  value, 
the  less  practical  the  investment  in  the  technol- 
ogies, some  of  which  are  relatively  expensive. 

Several  technologies  are  critical  to  the  introduction 
of  others. 

A methodology  that  could  reliably  induce 
estrus  synchronization  increases  the  economic 
feasibility  of  AI  and  embryo  transfer.  Likewise, 
the  refinement  of  embryo  storage  and  other 
freezing  techniques  would  advance  the  develop- 
ment of  those  technologies  still  being  developed, 
like  sex  selection  and  embryo  transfer.  Ad- 
vances in  in  vitro  fertilization  will  be  especially 
useful  to  a better  understanding  of  basic  repro- 
ductive processes  and  therefore  to  the  devel- 
opment and  application  of  the  more  speculative 
technologies. 

The  technologies  interrelate. 

All  the  technologies  combined  make  possible 
almost  total  control  of  the  reproductive  process 
of  the  farm  animal:  a cow  embryo  donor  may  be 
superovulated  and  artificially  inseminated  with 
stored,  frozen  sperm;  the  embryos  may  be  re- 
covered, then  stored  frozen  or  transferred  di- 
rectly to  several  recipient  cows  whose  estrous 
cycles  have  been  synchronized  with  that  of  the 
donor  to  insure  continued  embryonic  develop- 
ment. Before  the  transfer,  a few  cells  may  be 
taken  for  identification  of  male  or  female  chro- 
mosomes as  a basis  for  sex  selection.  Finally, 
two  embryos  may  be  transferred  to  each  recip- 
ient in  an  effort  to  obtain  twins.  (See  figure  33.) 

Techniques  not  yet  commercially  applicable 
all  require  embryo  transfer  in  order  to  be  use- 
ful. They  include  in  vitro  fertilization,  partheno- 
genesis, production  of  identical  twins,  cloning, 
cell  fusion,  chimeras,  and  rDNA  technology. 

The  technologies  described  in  this  section  are 
designed  to  increase  the  reproductive  efficiency 
of  farm  animals,  to  improve  their  genetic  merit, 
and  to  enhance  general  knowledge  of  the  repro- 
ductive process  for  a variety  of  reasons,  includ- 
ing concern  with  specific  human  medical  prob- 
lems, such  as  fertility  regulation  and  better 
treatments  for  infertility. 


Technologies  that  are  presently  useful 

SPERM  STORAGE 

The  sperm  of  most  cattle  can  be  frozen  to 
— 196°  C,  stored  for  an  indefinite  period,  and 
then  used  in  in  \'i\o  fertilization.  .Although 
many  of  the  sperm  are  killed  during  freezing, 
success  rates  [or  successful  conceptions  (table 
31)]  combined  with  other  adxantages  of  the 
technologies  are  enough  to  ensure  w idespread 
use  of  the  technology.  Short-term  sperm  stoi  age 
(for  one  day  or  so)  is  also  well-(le\eloped  and 
widely  used. 

The  major  advantages  of  storing  sperm  are 
the  increased  use  of  desirable  sires  in  breeding 
(see  figure  34),  the  ease  of  transport  and  spread 
of  desirable  germplasm  throughout  the  country 
and  the  world,  and  the  sa\  ings  fiom  slaughter- 
ing the  hull  after  enough  sp(>rm  has  been  col- 
lected. The  sperm  can  also  he  lest(*d  for  \cne- 
real  and  other  diseases  hefoi'(>  it  is  used  I hert'- 
fore,  the  use  of  sperm  banks  is  e.xpected  to  in- 
crease. Little  change  is  anticipated  in  .semen 
processing,  other  than  tin*  continued  refine- 
ment of  freezing  protocols,  which  dilfer  for 
each  species. 

ARTIFICIAI,  I,\SEM1\  VriOV 

The  manual  placcMuenl  of  speiin  into  tin* 
uterus  has  playcui  a ('cnlral  role  in  the  ilissemi- 
nation  of  \aluahl(?  g(‘rmplasm  thioughoul  the 
world’s  hertis  and  fhu'ks.  \ irtually  all  farm  spe- 
cies can  he  artificially  inst'minaled.  although  use 
of  the  technology  \ari(>s  widely  lor  different 
species— e.g.,  100  percent  of  the  Nation's  domes- 
tic turkeys  are  produced  via  AI  compan'd  with 
less  than  5 percent  of  beef  cattU*.  lA'en  hoiu'v- 

Table  31.  — Results  of  Superovulation  in 
Farm  Animals 


Average  number 

ovulations  normally  Number  of  ovulations 
expected  with  superovulatlon 


Cow 1 6-8 

Sheep 1.5  9-11 

Goat 1.5  13 

Pig 13  30 

Horse 1 1 


SOURCE:  George  Seidel.  Animal  Reproduction  Laboralor,  C'  ■ Siai»  u 
versily.  Fort  Collins.  Colo 


Ch.  9 — Advances  in  Reproductive  Biology  and  Their  Effects  on  Animal  Improvement  • 175 


\ 

Figure  33.— The  Way  the  Reproductive  Technologies  Interrelate 


N 

Recovered 

embryos 


Q o 


o 


Sexed? 


Frozen? 


Recipient  herd:  synchronized  estrus 
Embryo  transfer 
Each  get  two  for  twinning 


Calves 


Photo  Credit:  Science 


These  10  calves  from  Colorado  State  University  were  the 
result  of  superovulation,  in  vitro  culture,  and  transferto 
the  surrogate  mother  cows  on  the  left.  The  genetic 
mother  of  all  10  calves  is  at  upper  right 


SOURCE:  Office  of  Technology  Assessment, 


176  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


Figure  34. — Change  in  the  Potential  Number  of 
Progeny  per  Sire  per  Year  From  1939  to  1979 


Year 

SOURCE:  R.  H.  Foote,  Department  of  Animal  Science,  Cornell  University, 
Ithaca,  N.Y.,  unpublished  data. 


bees  and  fish  can  now  be  artificially  insemi- 
nated. 

It  pei'mits  the  widespread  use  of  germplasm 
from  genetically  superior  sires.  It  saxes  the 
farmer  the  cost  of  maintaining  his  own  sires  and 
is  valuable  in  disease  control,  especially  wlien 
germplasm,  rather  than  animals,  is  imported  or 
exported.  An  important  barrier  to  the  \\  ider  use 
of  Al,  especially  in  producing  beef  cattle,  is  the 
need  foi’  application  of  reliable  estrus  detection 
and  estrus  synchronization  technologies. 

An  expanded  role  for  Al  in  the  future  w ill  de- 
pend on  the  availability  of  accurate  information 
about  the  genetic  value  of  sperm  axailahle  for 
insemination.  A nationwide  information  system 
for  evaluating  germplasm  presently  exists  for 
only  one  species,  dairy  cattle. 

ESTRUS  SYNCHRONIZATION 

Estrus,  or  "heat,”  is  the  pei’iod  during  which 
the  female  will  allow  the  male  to  mate  vx  ith  her. 
The  synchronization  of  estrus  in  a herd,  using 
various  drug  treatments,  greatly  enhances  ,\l 
and  other  reproduction  programs. 


Federal  regulations  that  limit  tin*  ust'  of  pi-os- 
taglandins  or  progestog(‘ns  to  induc'c  .synchi'o- 
nized  estrus  in  horses  and  nonlactating  cows 
are  the  major  barrier  to  moix*  w idc’spix'ad  use  ol 
existing  technology. 

SUPEROVIH,  ATION 

SuperoMilalion  is  the  hormonal  stimulation  ol 
the  female,  resulting  in  the  rel(*ase  from  the 
oxary  ol  a largcM’  numbei'  of  oxa  than  normal 
(See  tabk'  31.)  (iomhiiK'd  xxith  \1  and  emhrxo 
transfer  of  th('  f(M'tiliz('d  oxa  into  surrogate 
mothers,  supei’oxiilated  oxa  can  result  in  the 
production  of  normal  otispring  xxith  tlu*  same 
rates  of  success  as  those*  folloxxing  normal  ox  il- 
lation. 

The  gri'ati'st  barrier  to  su|)erox  ulation  is  that 
the  d(?gi’(?e  of  suci'i'ss  cannot  he  |)redicted  for  an 
indix  idual  animal.  Otiu'r  harriers  include  xx  ideix 
x arving  (|ualitx  ol  hormone  batches  for  ox  ula- 
tion tri'atment,  Food  and  Drug  \dministration 
(Fl).\)  restrictions,  and  lack  of  d.ita  from  xx  Inch 
to  judgi*  the*  {'fleets  of  repeated  su|)erox  ul.ition 

In  the  future*,  inei'{*ase*el  unele*rstaneling  eil 
basie'  phxsieileigie'al  me'e  hanisms  xx  ill  l.ie  ilitale*  e*l- 
feirts  te)  impreixe*  the*  te*e  hnole)gx  II  h.is  .ideli- 
tieinal  e’eimnu're  ial  pe)te*nti;il  lor  she*e*p  anil  i .illli* 
hushanilrx',  einel  mui  h i urri*nl  e*llort  is  illre*e  ti*el 
loxx  ards  ele*x  e*loping  anil  ti'sling  a i'ommi*ri  i.il 
proi'i'diire*. 

i:\IHR\0  Ri.eox  I RX 

The*  abililv  to  {'olli'e  t te*rlili/e*el  oxa  Irom  the* 
ox  idui’ts  or  ute*rus  is  a ne*i  e*ssar\  ste*p  lor  1*111- 
brvo  transler  or  storage*  anil  lor  main  l■\pl-rl 
me*nts  in  ri*|)roilue  tix  e*  hiologx  I he*  li*e  hnolotix 
is  e*s|)e*e'ially  im|)ortanl  for  re*se*are  h into  proilui 
ing  ide*nlie*al  txx  ins.  pi'i  forming  i*mbrxo  hiopsie*-, 
for  se*\  {le*te*rminalion.  anel  olhe*r  pro|i*e  !•  ( 0111 
hilling  supe*rox  ulation,  artilie  ial  insi*minalioi! 
anel  e'liibryo  re*i  i)x  e*rx  maki*s  il  po-.sible*  to  l eil 
l(*ct  e'liibrx'os  from  a xiuinf;  hi*ile*r  helore  ri*.ie  h 
ing  |)ube*rty.  U he*n  some*  ilisorile*r  h.is  ilamai;i*{l 
the*  ox  ielui'ts  or  ute*rus.  I'liihrxo  re*i  e>xi*rx  Irom  a 
X aluable*  animal  make's  proi  ri'ation  po-.  able 

Both  surgii'al  anel  noiisurgiial  me-lhoels  .ire* 
e*urie*ntly  in  use*.  Surgii  al  ri*e  iixi*rx  is  ne*(  c-  ,ai 
for  shee'p,  goats,  anel  pig.s:  sue  h o|)i*r.itiom  are* 
limited  hy  the*  ele*xe*lo[)im*nt  ol  se  ar  tissue  \oc 


Ch.  9 — Advances  in  Reproductive  Biology  and  Their  Effects  on  Animal  Improvement  • 177 


surgical  embryo  reco\erv  is  pretei  red  for  the 
cow  anil  the  single  o\  ulation  of  the  horse.  The 
appmach  is  especially  important  in  dairy  cattle, 
since  it  can  he  performed  on  the  farm  without 
interrupting  milk  production. 

.\o  significant  aihtinces  i\in  he  ()i  eilictt‘d  foi’ 
the  immediate  future 

tAimtU)  1 It  \Nsi  lat 

Kmhryos  I'an  he  remoxed  from  one  animal 
and  implanted  into  the  o\  iiluct  or  utei'us  of 
another.  Ifoth  surgical  anil  nonsurgical  methods 
are  currentlx  in  use.  though  success  rates  of  the 
latter  are  much  loxx  er. 

The  technologx  can  obtain  offs|)ring  from  fe- 
males unable  to  support  a pregnancy,  increas- 
ing the  number  of  offs[)ring  from  xaluahle  fe- 
males and  introducing  nexx  geni‘s  into  patho- 
gen-free heiils.  Because  more  offspring  can  he 
obtained  from  the  donor,  undesirable  recessixe 
traits  can  he  ra[)iillx  iletected.  The  technologx  is 
also  useil.  along  x\  ith  short-  oi'  long-term  stoi  age 
of  the  emhrxos,  as  a means  of  trans[)orting 
germplasm  rathei'  than  the  xxhole  animal.  Cur- 
rent harriers  to  its  further  use  are  the  costs  in 
personnel  and  equipment,  especially  foi'  surgi- 
cal prix'edures.  anil  the  [)roxision  of  suitable 
recipients  for  a successful  transfer. 

The  use  of  embryo  transfer  should  increase 
in  the  future,  especially  xxith  animals  of  high 
xalue.  Nonsurgical  methods  xxill  increasingly 
replace  surgical  ones,  especially  for  coxvs  and 
horses.  .A  role  for  embryo  transfer  can  also  he 
predicted  in  progeny  testing  of  females,  obtain- 
ing txvins  in  beef  coxvs,  obtaining  progeny  from 
prepubertal  females,  and  in  combination  xxith 
in  X itro  fertilization  and  a xariety  of  manipula- 
tixe  treatments  (production  of  identical  txvins, 
selling  or  combining  ox  a from  the  same  animal, 
genetic  engineering). 

EMBRYO  STORAGE 

The  ability  to  store  embryos  increases  the 
adx  antages  of  embryo  transfer  procedures,  loxv- 
ers  the  cost  of  transporting  animal  germplasm, 
and  reduces  the  need  to  synchronize  estrus  in 
recipients.  It  xvill  also  be  important  in  the  study 
and  control  of  genetic  drift  in  animals. 


.Adequate  culture  sysliMiis  I'.xist  for  shoi  t-term 
storage  ot  embryos.  I hex  hax  e hiu’n  dex  eloped 
hx  trial-and-erior  anil  an*  not  optimallx’  di'fined 
lor  farm  specii's  at  presi'iit.  N'ex crtheless,  coxx' 
embryos  haxe  been  stored  for  'A  days  in  the  tieil 
ox  iduct  of  a rabbit. 

I.ong-term  storage,  or  freezing  of  embryos, 
exists,  hut  protocols  nei'd  to  he  impi'oxinl.  As 
manx  as  txx o-thii’ds  of  the  stored  embryos  liie 
xxith  [jresent  methods,  lloxvever,  for  some  uses 
embryo  freezing  is  already  pi’ofitahle. 

In  the  luture,  tlu‘  di'xelopment  of  prei'ise  em- 
hrxo  cultui'e  technolog^x'  xxould  help  the  dexel- 
opment  of  all  ti'chnologies  inxoixing  the  pro- 
longed manipulation  of  gameti!s  and  embryos 
outside  the  reproductix c tract.  lA I'ntually,  as 
freezing  technology  improxes,  ni^arly  all  em- 
hi-yos  taki'n  from  cattle  in  North  .AnuM'ica  xvill 
he  stored,  rather  than  transferred  immediately. 
It  appears  that  emhi’X'os  successfully  storeil  xx  ill 
surxixe  foi-  sexi’ial  centuries  and  possibly  foi- 
millenia. 

SE.\  SELECTION 

rhe  ability  to  di’termine  the  se.x  of  the  un- 
born, or  of  sperm  at  fertilization,  xxill  have  nu- 
merous |)ractical  and  experimental  applications. 
The  most  reliable  method  is  karyotyping,  by 
means  of  xx  hich  nearly  txx'o-thii'ds  of  embryos 
can  he  sexed.  Another  method,  xvhich  tries  to 
identify  sex-specific  pi’oilucts  of  certain  genes, 
is  under  dexelopment.  A reliable  method  for 
separating  male-producing  sperm  from  female- 
producing  sperm  has  not  been  achieved,  though 
sex  eral  patents  are  held  on  x arious  tests  of  this 
type. 

Before  any  method  has  any  practical  effect  on 
the  production  of  farm  animals  it  must  become 
simple,  fast,  inexpensix  e,  reliable,  and  harmless 
to  the  embryo.  The  present  state  of  the  art  is 
largely  a consequence  of  research  in  male  fertil- 
ity and  in  sperm  survival  after  frozen  storage. 

TWINNING 

Twins  can  be  artificially  induced  by  using 
either  embryo  transfer  or  hormonal  treatments. 
The  first  approach  is  more  effective.  Selection 
among  female  sheep  for  natural  twin  produc- 


178  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


tion  has  been  very  rewarding,  while  selection 
for  twinning  in  other  species  has  not  received 
much  attention. 

Twinning  in  nonlitter-bearing  species  would 
greatly  improve  the  feed  conversion  ratio  of 
producing  an  extra  offspring.  The  most  impor- 
tant barriers,  besides  the  high  cost  of  embryo 
transfer  techniques,  include  extra  attention 
needed  for  the  dam  during  gestation,  parturi- 
tion, and  lactation. 

More  speculative  technologies 

IN  VITRO  FERTILIZATION 

The  manual  joining  of  egg  and  sperm  outside 
the  reproductive  tract  has,  for  some  species, 
been  followed  by  successful  development  of  the 
embryo  through  gestation  to  birth.  The  species 
include,  at  this  writing,  the  rabbit,  mouse,  rat, 
and  human.  Consistent  and  repeatable  success 
with  in  vitro  fertilization  in  farm  species  has  not 
yet  been  accomplished.  The  cases  of  reported 
success  of  in  vitro  fertilization,  embryo  reim- 
plantation, and  normal  development  in  man  are 
beginning  to  be  documented  in  the  scientific 
literature. 

The  in  vitro  work  to  date  has  attempted  to  de- 
velop a research  tool  so  that  the  physiological 
and  biochemical  events  of  fertilization  could  he 
better  understood.  Despite  the  wide  public  at- 
tention it  has  received  in  the  recent  past,  the 
technology  is  not  perfected  and  will  have  little 
practical,  commercial  effect  in  producing  in- 
dividuals of  any  species  in  the  near  future. 

Practical  applications  would  include:  a means 
of  assessing  the  fertility  of  ovum  and  sperm;  a 
means  of  overcoming  female  infertility  by  em- 
bryo transfer  into  a recipient  animal;  and,  when 
coupled  with  storage  and  transfer,  a means  of 
facilitating  the  union  of  specific  ova  and  sperm 
for  production  of  individual  animals  with  pre- 
dicted characteristics. 

Many  of  the  practical  applications  should  h(!- 
come  available  within  the  next  10  to  20  years. 
Further  development,  along  with  the  storage  of 
gametes,  should  allow  fertilization  of  desired 
crosses.  This  technology  may  he  combined  with 
genetic  engineering  and  sperm  sexing  in  the 
more  distant  future. 


PARTHENOGENESIS 

Parthenogenesis,  or  “virgin  birth,”  is  th<>  ini- 
tiation of  dex  elopment  in  the  absence  of  s|)ei'm. 
It  has  not  been  demonstrated  or  descrilu'd  lor 
mammalian  species,  and  the  best  a\  ailahle  infor- 
mation indicates  that  the  maintenance  of  pai'- 
thenogenetic  de\elo|)ment  to  [)i'oduce  normal 
offspring  in  mammals  is  pi'esently  im|)ossihl(‘. 

CIAINING 

The  possibility  of  protliu'ing  gcMU'tically  iden- 
tical indixiduals  has  fasc'inatc'd  l)oth  scientists 
and  the  general  public.  .As  tar  as  liwstock  are 
concerned,  theix^  ai'(?  scnc’ial  ways  to  obtain 
genetically  identical  animals.  rlu>  natui'al  way  is 
through  identical  tw  ins,  although  these  are  rai  l* 
in  species  other  than  cattk’,  sh(>ep,  ami  pri- 
mates. Foi’  pi'actic'al  purposes,  highly  inbred 
lines  of  soim?  mammals  are  ali  ('ady  considered 
genetically  identical;  first  generation  crosses  ol 
these  lines  are  also  considered  genetically  iden- 
tical and  do  not  suffer  Irom  the  depressive  el- 
fect  of  inhrc'eding. 

hahoratory  imnhods  for  producing  clones  in- 
clude div  iding  early  embryos.  I he  results  ol  re- 
cent e.\|)('rim('nts  in  the  production  of  identical 
offspring  using  these*  t(*chni(|ues  are  shown  in 
table!  32. 

,\noth('i'  methodology  involves  the  insertion 
of  the*  nucleus  of  one  cell  into  another,  either 
liefore!  oi’  alter  the  original  genetic  complement 
of  the!  "re!e'e*iv  e*r”  e e*ll  is  deslroyeef  Iteseare  hers 
have!  feuind  in  ce'itain  amjihihia  that  nucle.ir 
trans|)lantation  fiom  a body  cell  ol  an  embryo 
into  a zvgote  e an  le*ael  to  the  elev  elopmeni  ol  a 
sexually  mature*  I reig. 


Table  32.  — Experimental  Production 
of  Identical  Offspring 


Methodology 

Result 

Dividing  2-cell  embryo  in 
half 

1 pair  identical  mouse  twins 

Dividing  morulae®  in  half 

8 pairs  of  identical  mouse 
twins 

Dividing  2-cell  embryos  in 

5 pairs  of  identical  sheep 

half 

twins 

Dividing  4-cell  embryos  in 

1 set  identical  sheep 

four  parts 

quadruplets 

^An  embryo  wilh  16  lo  50  cells;  resembii--.  a muitxf', 

SOURCE.  Beniamin  G Bracken.  School  ol  Volenna,,  Mr  ' , r ,>  ,,c  -.  • 


Pennsylvania.  Kennell  Souarc  Pa 


Ch  9 — Advances  in  Reproductive  Biology  and  Their  Effects  on  Animal  Improvement  *179 


rlu'  itltMl  tt‘clmi(|iu'  li)i-  making  genetic  (•t)|)ii*s 
of  an\  atliilt  mammal  iinoKcs  ins<M'tin^ 

tin*  lUK'ltms  from  a hody  roll  (oof  a sr\  rrll)  f Vom 
an  adult  indi\  itlual  into  an  o\  um.  \rhir\  iof'  this 
will  prohahK  takr  yrars,  if  indrrd  it  is  possihir 
at  all.  sinrr  thrrr  is  somr  r\  idrnrr  that  most 
atlulf  l)(ul\  rolls  arr  iiTo\  tM'sihly  dif  trr(*ntialrd. ' 

Sri  ioiis  trrhniral  hai  rirrs  must  hr  o\  t‘rromr 
hrf'orr  acKanta^rs  in  animal  produrtion  ran  h(‘ 
f'orrsrrn. 

eta. I,  f i SION 

This  ti‘rhnolo^\  fusrs  two  maturr  o\ a or  fri'- 
tilizrs  onr  o\um  w ith  anothrr.  C'omhining  o\a 
from  ihr  samr  animal  is  ralird  srlfin^. " I'lu* 
comhination  of  o\a  has  rrsulfrtl  in  \ rry  rarly 
dr\  rlopnu*nt  of  tht'  Iransfrrrrd  rmhr\'o.  hut  no 
furthrr  dr\ riopmrnt  has  h(>rn  ia*poi  trd. 

C!rll  fusion  trrhnolo^\  ma\  somrday  pro\r 
usrful  for  transfrrrin^  firnotir  matrrial  fiom  a 
somatir  rrll  into  a frrtili/rd  singir-ri'll  rmhryo 
foi’  thr  [)urposr  of  rioning.  Sidfin^  would  rapid- 
ly rrsull  in  purr  grnrlir  (inhi  rd)  linos  for  usr  as 
brooding  storks.  I hi'  trrhni(|ur  rould  also  load 
to  thr  rapid  idrntifiration  of  undrsii'ahlr  rrrrs- 
si\r  traits  that  rould  hr  eliminated  from  the 
species. 

CHIMtlK  \S 

The  produrtion  of  chimeras  requires  the  fu- 
j,  sion  of  two  or  more  early  emhr\os  or  the  addi- 


Mn  Jamuirv  1981  it  was  rt'portt'd  that  Ixxlv  cells  from  a very 
early  enthryo  could  act  as  donors  of  nuclei  for  cloned  mice. 


tion  of  extra  cells  to  blastocysts,  fhese  genetic 
compoiKMits  may  he  from  closely  relatful  Init  dif- 
ferent sprci(*s. 

t,i\f‘  chimeras  between  two  species  of  mouse 
ha\('  hr(Mi  produced.  tlowe\(M',  practical  appli- 
cations of  chimera  tc'chnology  to  li\  estock  are 
not  oh\  ious  at  this  stage  of  tlex  elopment.  fhe 
main  object i\e  of  this  research  is  to  provide  a 
genetic  tool  for  a better  understanding  of  de\  el- 
opmiMit  and  mat(>rnal-fetal  interactions. 

lU'COMHIV  AN T l)N  A AM)  (JliNK  TKANSFEH 

The  nuH'hanics  of  diriH'tly  manipulating  the 
l)\.\  molecules  of  farm  animals  ha\e  ivil  yet 
lu'rn  workful  out.  However,  cells  from  mice 
ha\c  luMMi  mixed  with  pieces  of  chromosomal 
l).\  A,  w hich  became  stably  associated  with  the 
cells'  own  I).\.\.  In  addition,  on  September  3, 
19<S(),  th(>  successful  introduction  of  foreign 
I)\  A into  mouse  embryos  was  announced.  The 
embryos  wei'r  implanted  into  surrogate  moth- 
ers who  ga\e  birth  to  mice  containing  altered 
D.VA.  \\  hether  or  not  the  l)\V\  was  active  is  un- 
known at  this  writing. 

Knowledge  of  the  genetics  of  farm  animals 
must  improx  e before  rDN'A  or  other  gene  trans- 
fer methods  will  he  of  practical  benefit  in 
producing  meat  and  lix  estock  products.  Before 
genes  can  he  altered  they  must  he  identified, 
and  gene  loci  on  chromosomes  must  be 
majiped.  Work  toxx  ard  this  goal  has  begun  only 
recently  and  rapid  progress  cannot  be  antici- 
pated. Multixariate  genetic  determinants  of 
characteristics  are  anticipated  to  be  the  rule. 


Genetics  and  animal  breeding 


I Txvo  characteristics  distinguish  the  reproduc- 
tion of  farm  animals  from  that  of  single-cell  or- 
! ganisms:  animal  reproduction  is  sexual— male 
and  female  germ  cells  must  be  brought  together 
to  initiate  pregnancy  and  produce  offspring; 
and  animal  reproduction  is  sloxver  (the  genera- 
tion interx'al  is  longer),  thus  the  economic  bene- 
fits of  specific  gene  lines  may  take  years  to  be 
captured.  These  txvo  characteristics  limit  the 
speed  and  extent  to  xvhich  genetic  improve- 


ments can  be  made.  Reliable  information  about 
the  genetic  x alue  of  particular  individuals  is  the 
key  to  overcoming  limitations,  for  it  can  simpli- 
fy specific  breeding  decisions  and  spread  desir- 
able genes  throughout  the  Nations’s  herds  and 
flocks. 

The  use  of  applied  genetics  for  farm  species  is 
indirect.  Breeders  do  not  work  with  individual 
genes;  rather,  they  must  accept  a genetic  pack- 


180  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


age  that  includes  both  beneficial  and  harmful 
traits.”  The  breeder’s  most  important  capital  is 
embodied  in  the  animals  with  which  he  works. 
To  upgrade  this  capital,  to  increase  the  genetic 
\alue  of  his  hi'eeding  stock,  the  breeder  must 
have  reliable  information  on  the  genetic  value 
of  the  gernifilasm  he  is  considering  introducing. 
Since  an  individual  farmei'  usually  does  not 
have  the  resources  to  collect  and  process  data 
on  |3erformance  of  indiv  iduals  outside  his  own 
herds,  he  must  tui'u  to  outside  sources  of  infor- 
mation when  deciding  which  new  germplasm  to 
introduce. 

The  i'ec|uirements  of  such  an  information  sys- 
tem ai'e  extensive.  In  the  United  States  today, 
only  one  such  system  exists.  The  National  Co- 
operative  Dairy  Hei'd  Improvement  Program 
(N(d)HIP)  is  a model  program  that  could  he 
adapted  to  other  s|)ecies  where  the  benefits 
from  adv  anced  technologies  would  he  enhanced 
by  availability  of  populationwide  data. 

The  National  Cooperative  Dairy  Herd 
Improvement  Program 

Over  the  past  50  years,  the  U.S.  dairy  indus- 
try has  used  test  records  of  individual  animals 
to  help  in  breeding  decisions.  NCDHIP  is  a na- 
tionwide program  for  collecting,  analyzing,  and 
disseminating  information  on  the  performance 
of  dairy  cattle.*^  It  is  the  result  of  a memoran- 
dum of  understanding  among  Federal  and  State 
agencies,  local  dairymen,  and  industry  groups 
across  the  United  States. 

In  NCDHIP,  local  Dairy  Herd  Improvement 
Association  (DHIA)  officials  go  to  the  dairies  to 
collect  the  performance  data  on  indiv  idual  ani- 
mals. These  data  then  become  part  of  the  Ofji- 
cial  Dairy  Recordkeeping  Plans.  The  data  are 
standard  for  all  participating  herds  across  the 
United  States.  They  are  sent  to  the  Animal  Im- 
provement Programs  Laboratory  (AIPL)  at 
USDA  in  Beltsville,  Md.,  which  analyzes  them 
and  incorporates  them  into  the  “USDA-DHIA 

''Philip  Hiuiciler,  Biology  and  the  Ftilure  of  Man  (New  Voi  k:  0,\- 
lord  Unix  er.sil V Pres.s,  i;)70l,  |)|j.  sriS-SST. 

'^Kor  a complete  hi.slorv  ol  perldrmance  le.stitif'  of  dairy  catlli? 
in  the  United  State.s,  .see:  Gerald  .1.  Kiiif',  I'he  National  Cooperative 
Dairy  Herd  Improvement  Program,  Dairy  Herd  lm|)rovemeni  I.et- 
ter  49,  No.  4,  Jnlv  1973,  USD,\,  ,VRS. 


Sire  Summary  List,”  published  hiannually. 
These  summaries  are  public  information. 

In  addition  to  the  official  plan,  N(d)HlP  also 
includes  several  unofficial  plans,  which  have 
less  stringent  regulations  for  data  collection  but 
which  offer  each  dairyman  a comparison  of  bis 
herds  with  other  herds  across  the  Nation.  The 
results  of  unofficial  plans  are  not  intended  to  he 
used  as  guidelines  for  selecting  germplasm  from 
outside  one’s  herd. 

The  following  characteristics  conti'ibut(>  to 
NCDHIP's  success: 

• It  is  a cooperative  program;  in)  group  or  in- 
dividual is  forced  to  i)articipat(v  NcniM  the- 
less,  it  has  successfully  brought  togfMher 
individuals.  State  and  Ff'deral  agencies, 
breed  associations,  and  professional  and 
scientific  socicfties  for  the  [lui'suit  of  a com- 
mon goal.  It  is  almost  totally  financed  by 
the  dairymen  tlKMiiselv (>s.  In  the  national 
coordinating  gi'oup,  all  those  v\  ith  an  intei'- 
est  in  the  industrv  haw  a voice  in  formu- 
lating policy  for  the  jfiogram. 

• It  is  flesible;  a dairyman  can  use  the  jhm  - 
formance  I'f'cords  from  the  unofficial  plans 
to  evaluate  tlu'  animals  within  his  herd  or 
he  can  turn  to  the  official  sire  summai  i«*s 
to  make  comparisons  with  participating 
herds  throughout  the  .Nation.  Ihe.se  data 
ai'e  us('ful  both  for  com|)aring  the  perlorm- 
ance  of  one’s  herd  and  bri'ed  with  others 
and  foi'  selecting  nmv  germplasm  lor  in- 
troduction into  the  herd 

• Its  data  ai'e  regarded  as  impartial;  disinterest 
on  th('  part  of  the  local  DHIA  otiicial  who 
collects  the  data  and  the  high  securitv  sur 
rounding  the  processed  intormation  .ire 
central  to  the  program's  success  \IPLs 
analyses  and  sire  summaries  are  respected 
both  nationally  and  inti'rnation.illv  m no 
small  part  because  of  freedom  Irom  (om- 
men'ial  [iressures. 

Ap|)roximately  5(), ()()()  herds  w ith  .ilmost  2 K 
million  cows  were*  enrolled  in  the  otlici.il  jil.ins 
of  N(M)HIP  in  In  each  ol  IS  vc.irs  rec orded 

Ix^tweiMi  HHil  and  H)7S,  cows  enrolled  m the 
()ffic:ial  Dairv  Recordki*e[)ing  Pi. ins  m \(  DIIIP 


Ch.  9— Advances  in  Reproductive  Biology  and  Their  Effects  on  Animal  Improvement  • 181 


4 000  11)  ot  milk.  |)t>r  lai'tation.  In  llie  testinfi  \ t‘ai' 
(1077-7iS),  the  suptM'iority  sur[)assecl  5,000  II)  pt>r 
I'DW  This  5,000-11)  siipei'iority  represents  52 
peri'ent  more  milk  per  laetation.  1 ht>  inereases 
in  protliietion  ()t‘r  eow  I'tvsiilt  trom  improx  ement 
in  both  management  teehni(|iies  and  genetic 
producing  ahilitx . 

SextM'al  factors  intluence  the  I'ates  of  pai  tic- 
i[)ation  in  the  XCDllll’  from  State  to  Statt\  from 
region  to  I'egion,  and  from  breed  to  breed.  In 
some  States,  expansion  of  NCDHll’  memhei'ship 
is  not  a high  prioritx  of  tin*  Statt*  C'oopiM'atix e 
Kxtension  St*r\  ice.  In  some  ai’eas.  the  I'elatix  e 
im[)ortance  of  dairx  ing  as  an  ent(M‘prise  is  low ; 
therefore,  a strong  local  1)111  \ organization 
does  not  exist  l,ikex\is(‘,  in  aia'as  where  daii'x- 
ing  is  a part-time  operation,  daii ymen  have  less 
time  and  initiatixe  for  partici[)ating  in  the  pro- 
gram (although  many  [)artici|)ate  iti  .\t4)HIF’s 
unofficial  plans).  W here  dairymen  rely  on  their 
oxx  II  hulls  and  use  little  W in  breeding,  progeny 
testing  is  extremely  limited.  ,\o  single  factor 
causes  dairymen  in  .some  States  to  take  greater 
adx  antage  of  the  su[)erior  germplasm  ax  ailahle 
to  them  rhe  importance  of  strong  national 
leadership  cannot  he  ox  eremphasized  in  ex- 
plaining the  great  differences  among  breeds  in 
participation  rates.  (See  table  33.)  Farsighted 
leadership  played  a large  role  in  dex  eloping  the 
genetic  gain  of  Holsteins,  xx  hich  represent  90 
percent  of  the  I .S.  dairx  herd  today. 

The  genetic  gains  resulting  from  XCDHIP  are 
inipressixe,  suggesting  a model  for  spreading 
genetic  superiority  throughout  the  Nation’s 
other  herds.  XCDHIP  also  shoxx  s the  importance 

Table  33.— National  Cow-Year  and  Averages  for 
All  Official  Herd  Records,  by  Breed 
May  1,1 978- Apr.  30,1979 

Cow-years 


Breed (#)  Milk  (lb)  Fat(%)  Fat  (lb) 

Ayrshire 17,135  11,839  3.96%  469 

Guernsey 57,577  10,858  4.64  504 

Holstein 2,297,684  15,014  3.64  547 

iJersey 89,449  10,231  4.90  501 

'Brown  Swiss 24,247  12,368  4.04  500 

Milking  shorthorn  2.130  10,451  3.65  381 

Mixed  and  others.  83,139  13,077  3.80  497 


iSOURCE:  U S.  Department  of  Agriculture.  Science  and  Education  Administra- 
tion, Dairy  Herd  Improvement  Letter  55.  #2.  December  1979.  pp.  5-6. 


ot  combining  reliable  ex  aluation  of  germplasm 
xxith  the  use  of  reproductixe  technologies, 
rhese  technologies  art?  of  only  academic  in- 
terest XX  hen  thex'  are  used  alone;  it  is  xx  hen 
superior  germplasm  can  he  spread  throughout 
the  Nation  that  the  .American  consumer 
benefits. 

Othi^r  spei'ies 

Progeny  testing  schemes  for  other  species  are 
not  as  dexeloped  as  tht?y  are  foi'  dairy  cattle. 
There  ai'e  sexeral  reasons  for  this  lack  of 
testing: 

• Difjiculty  in  establishing  a selection  objective 
around  which  to  design  a testing  program. 
.Milk  x'ield  and  fat  content  xvere  ohxious 
traits  for  selection  in  dairy  cattle.  Other 
species  hax  e no  such  simple  traits  for  selec- 
tion. It  has  been  ohserx  ed  that,  “The  lack  of 
definition  of  economic  selection  ohjectixes 
in  a precise,  soundly  based  manner  is  one 
of  the  serious  xxeaknesses  of  much  animal 
breeding  of  the  past.’’’^ 

• Differences  in  management  systems.  Artifi- 
cial insemination  is  essential  to  the  intro- 
duction of  superioi'  germplasm;  where  it  is 
difficult  to  practice  Al,  elaborate  testing 
schemes  are  not  useful— e.g.,  in  the  Na- 
tion’s beef  herds,  progeny  testing  will  have 
to  await  more  widespread  use  of  AI. 
Though  sxvine  are  increasingly  raised  in 
confined  housing  systems,  poor  fertility  of 
boar  sperm  after  freezing  and  thawing  and 
heat  detection  difficulties  have  limited  the 
use  of  AI. 

• Conflicting  commercial  interests.  Beef  bulls, 
for  example,  continue  to  be  sold  to  some 
extent  on  the  basis  of  fancy  pedigrees  and 
lines,  with  relatively  little  objective  in- 
formation on  their  genetic  merit.  Although 
some  genetic  improvement  programs  now 
exist,  the  beef  breed  associations  may  not 
support  interbreed  comparisons  because 
some  breeds  would  show  up  poorly. 

• Conflicts  between  short-  and  long-term  gains. 
Cross-breeding  for  the  benefits  of  hybrid- 


'^L.  E.  .A.  Rouson,  "Techniques  of  Livestock  Improvement,"  Out- 
look on  Agriculture  6:108,  1970. 


182  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


ization  is  particularly  attractive  to  owners 
of  commercial  herds  and  flocks  who  con- 
stantly replace  their  stocks.  This  genetic 
improvement  is  noncumulative— the  im- 
provement does  not  continue  from  gener- 
ation to  generation.  At  present,  no  strong 
interest  exists  for  improving  the  Nation’s 
heef  herd  as  a whole,  and  the  individual 
breeder  cannot  effectively  evaluate  the 
germplasm  available  to  him. 

Swine. — There  is  no  Nationwide  testing  pro- 
gram for  hogs  in  the  United  States.*  However,  a 
study  of  needed  research  prepared  by  the 
USDA  in  1976  noted  that  the  production  rate  of 
approximately  13  pigs  marketed  per  sow  per 
year  in  the  United  States  could  be  significantly 
improved.  The  biological  potential  is  at  least  20 
to  25  pigs  per  year.  Similarly,  a successful 
breeding  program,  along  with  other  managerial 
changes,  could  reduce  the  fat  and  increase  the 
lean  content  of  pork  by  as  much  as  10  to  15  lb 
per  carcass. 

The  ARS  study  noted  that  “.  . . an  area  that 
warrants  particular  attention  is  the  develop- 
ment of  a comprehensive  national  swine  testing 
program  leading  to  the  identification,  selection, 
and  use  of  genetically  superior  boars,  together 
with  guidelines  for  the  development  and  use  of 
sow  productivity  and  pig  performance  in- 
dexes.”''* In  the  case  of  swine,  the  increased  use 
of  intensive  housing,  which  allows  reproductive 
control,  should  increase  the  impetus  for  prog- 
eny testing.  Likewise,  pinpointing  areas  where 
considerable  improvement  remains  to  be  made 
should  lead  to  the  identification  of  selection 
objectives. 

Beef. — After  World  War  II,  a few  breeders 
became  increasingly  interested  in  problems  of 
inbreeding  and  the  economic  costs  of  dwarfism. 
By  that  time,  some  had  been  trained  in  genetics 
and  some  breed  associations  and  State  agencies 
initiated  localized  testing  programs  for  these 
traits.  In  1967,  a "Beef  Improvement  Federation” 


‘There  are  several  State  programs — in  Indiana,  North  Carolina, 
and  Tennessee.  Some  of  these  programs  may  test  only  growth  and 
not  litter  size. 

'■•U.S.  Department  of  Agriculture,  Agricultural  Research  Ser\  - 
ice,  ARS  National  Research  Program,  Swine  Production,  NKI’  No. 
20370,  October  1976. 


of  local  and  breed  groups  was  formed  to  try  to 
consolidate  the  different  systems  of  the  State 
improvement  programs.  The  Federation  is  now 
involved  in:'® 

• establishing  uniform,  accurate  records, 

• assisting  member  organizations  in  de\  elop- 
ing performance  programs, 

• Encouraging  cooperation  among  all  seg- 
ments of  the  industry  in  using  records, 

• Encouraging  education  by  emiihasizing  the 
use  of  records, 

• developing  confidence  in  performance  test- 
ing throughout  the  industry. 

Despite  these  efforts,  only  about  3 pi'rcenl  of 
beef  cattle  nationally  are  recoi'ded.  This  rel- 
atively low  participation  rate,  \\  1umi  compared 
with  NCUHIF,  has  both  a Uu  hnological  and  an 
institutional  explanation.  Unck'r  th(’  larg(*l\  (‘x- 
tensi\  e heef  raising  system  in  th(»  Uniti'd  Stale's, 
AI  is  difficult  as  long  as  estrus  detection 
technologies  are  una\ailahle.  Natural  stud  se>r\- 
ice  is  usually  moi'e  economical.  Institutional  har- 
riers also  pre\ent  the  d(‘\ I'lopment  of  a strong 
genetic  evaluation  program— e.g.,  the  hie'ed 
associations  are  not  all  eager  to  ha\(*  theii' 
breeds  consistently  compared  u ith  othe'i  s.  Uk«*- 
wise,  some  owners  of  hulls  for  stud  s»*r\  ice 
would  lose  business  in  a strict  testing  scheme. 

Goats. — I hough  little  genetic  work  has  been 
done  on  goats  in  the  past,  the  dairy  goat  in- 
dustry has  heconu'  moic  \ isihle  in  the  past  h’w 
years.  I'he  desire'  of  goat  hre'e'de'i  s to  particip.ite 
in  NCDllll’  l('d  to  the'  formation  nl  a ( ooi - 
dinating  Sul)-(irou[)  for  Dairy  (lOats  \ i ('\  lew  ol 
the  I'eseai'ch  pe'rfoi  iiK'd  indicate'd  a gi  i'.it  need 
foi’  resf'ai’ch  in  almost  e\  ('l  y are'a  ol  production 
As  a result,  .All’l,  d('\('loped  a plan  lor  a genetic 
impro\ement  piogram.  I he  leadi'i  ship  in  the 
dairy  goat  induslrx'  was  coin  inced  that  it  i ould 
attain  gene'tic  impro\  I'liK'nl  taster  .ind  at  a 
lower  ('ost  \ ia  N( Dllll’  than  it  could  lor  an\ 
other  type'  of  re'se'an  h. 

In  1979,  .All’l.  r('cei\('el  a .Sl5.()t)()  gi.inl  Imm 
the  Small  I'arms  Be'search  I unelinf;  to  support 
the  ele'\'e'le)pme'iit  of  genetii'  evaluation  priMi- 

'■'R  I.  Willh.mi,  l.cnrlK  \ili\ilx  in  tin  I s i..,  i 
Journal  I’iipcr  Nl)  J-792.1  nl  Ihi-  Inw.t  ii  iillo' ,ii  .n  all  '■  < I • 
nnmiis  l.xpcrimi'nl  St.ilion  Vinrs  low.i  pinii  il\i>  • a 


Ch.  9 — Advances  in  Reproductive  Biology  and  Their  Effects  on  Animal  Improvement  • 183 


idiii’es  goats.  (U'lietif  evaluations  tor  yield  of 
: dairy  goat  hueks  w ill  he  av  ailable  hetore  the  end 
|Ot  fiscal  year  1980.  Bt'cause  limited  genetic  im- 
provement tor  yielil  has  occui  red  in  tlairv  goats 
in  the  past  thest'  evaluations  will  prohahly  have 
|a  sigtiit'icant  im[)act  on  the  industry.  .\ll’l,  can 
V irtually  guarantee  htMieticial  results  because  of 
.the  ilata  available  from  .XCDlllP.  its  own  e.xper- 
ti.se  in  genetics,  statistics,  and  com|)ut(>r  tech- 
tiolog^v . and  the  dt'cades  ot  highlv  effectiv  e re- 
search on  genetic  imf)rovement  of  dairy  cattle 
that  can  he  aila[)ted  for  the  dairv  goat  industry. 
HowevtM'.  fuiuling  for  the  goat  testing  program 
'ivmains  on  a v ear-to-v  ear  basis. 

! CONCl.l  SION 

' .\('l)tllP  has  show  II  how  im|)ortant  genetic  in- 
'formation  is  to  tl«‘  production  of  meat  and  dairy 
products.  The  obstacles  to  such  a pi’ogram  ai'e 
also  formidable,  hut  every  failure  to  capitalize 
on  genetic  potential  is  paid  foi'  by  .American 
consumers.  It  has  also  show  n that  w here  selec- 
tion objectiv  es  can  he  identified  and  agreed  on, 
and  w here  conflicting  interests  can  he  brought 
itogether  to  develop  a [)rogram  serving  all  in- 
Iteresls.  genetic  improv  ement  can  become  a cen- 
tral objective  in  breeding  programs  across  the 
jcountry.  Without  reliable,  evaluative  data  on 
breeding  stock  the  Nation  s breeders  will  have 
little  interest  in  adopting  new  breeding  technol- 
ogies as  they  become  av  ailahle. 

Impacts  on  breeding 

j An  improvement  in  germplasm,  like  an  in- 
crease in  the  nutritional  content  of  fertilizer  or 
new  and  improv  ed  herbicides  and  pesticides,  in- 
'Creases  the  quality  of  the  physical  capital  used 
•on  the  farm.  It  is  likely  that  much  improvement 
jean  still  be  made  in  the  germplasm  of  all  major 
farm  animal  species  using  existing  technologv’. 

1 Selecting  for  desired  characteristics  causes  a 
Ispecific  qualitativ  e change;  it  enhances  the  effi- 
jciency  of  the  information  contained  w ithin  each 
cell.  The  genetic  information  in  each  cell  of  a 
farm  animal  is  either  more  or  less  desirable  or 
iOfficient  than  information  in  the  cells  of  another 
animal,  depending  on  how  it  performs  on  im- 
<portant  traits.  Superior  germplasm  can  be  used 
in  breeding  decisions  to  upgrade  a farmer’s 


breeding  or  producing  stock.  (DHIA  programs 
are  the  best  example  of  how  information  might 
be  distributed.) 

Resources  invested  in  genetics  and  in  technol- 
ogies related  to  genetics  will  have  high  payoffs— 
e.g.,  in  a classic  study*®  of  the  payoff  to  research 
in  hybrid  corn  and  in  subsequent  studies  of 
other  types  of  genetic  improvement,  a high 
costAjenefit  ratio  for  such  research  was  found. 
The  original  study  also  show'ed  that  the  absolute 
market  value  of  a particular  product  is  an  im- 
portant factor  influencing  the  rate  of  return  on 
a given  research  expenditure.  In  general,  the 
greater  the  aggregate  value  of  the  product,  the 
greater  the  rate  of  return  on  a research  expend- 
iture.'^ Thus,  the  large  expenditures  for  meat 
and  animal  products  in  the  United  States  sug- 
gest a great  payoff  in  applied  genetic  research. 
Beef  purchases  alone  account  for  between  2 and 
5 percent  of  the  American  consumer  dollar,  and 
the  total  maii<et  value  for  beef  is  more  than 
twice  that  for  corn  in  the  United  States. 

DAIRY  CATTLE 

Total  milk  production  has  been  stable  for 
many  years.  W hile  milk  production  per  cow  has 
gone  steadily  upward,  the  number  of  cows 
during  the  past  35  years  has  decreased  propor- 
tionately. (See  figure  29.)  Milk  production  per 
cow  should  continue  to  increase,  assuming  that 
no  radical  changes  in  present  management  sys- 
tems occur.  The  increase  in  production  per  cow 
could  continue  even  if  no  bulls  superior  to  those 
already  available  are  found,  simply  as  a result  of 
more  farms  switching  to  existing  technology 
and  existing  bulls.  Moreover,  bulls  produced 
from  this  system  are  increasing  in  superiority. 

The  number  of  dairy  cows  calved  as  of  Janu- 
ary 1,  1980,  was  10,810,000.  It  has  remained  rel- 
ativ'ely  stable  for  the  past  year,  but  may  de- 


■®Zvi  Griliches,  "Research  Costs  and  Social  Returns:  Hybrid  Corn 
and  Related  Innovations,  "Journal  of  Political  Economy  66:419,  Oc- 
tober 1958.  See  also  R.  E.  Evenson,  P.  E.  VV'aggoner,  and  V'.  VV.  Rut- 
tan,  "Economic  Benefit  From  Research:  An  Example  From  Agricul- 
ture," Science  205:1 101,  Sept.  14,  1979. 

■nv.  Peterson  and  Vujino  Hayami,  "Technical  Change  in  Agricul- 
ture," Staff  Papers  series  No.  DP73-20,  Department  of  Agriculture 
and  Applied  Economics,  Uni\ersity  of  Minnesota,  St.  Paul,  Minn., 
July  1973. 


184  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


crease  to  around  10  million  in  the  next  decade  if 
milk  production  continues  to  increase. 

Artifical  Insemination. — An  example  of 
the  interaction  between  technologies  and  genet- 
ic improv'ement  is  shown  in  table  34.  The  “pre- 
dicted difference”  (PD)  in  milk  production  rep- 
resents the  ability  of  individual  bulls  to  genet- 
ically transmit  yield— the  amount  of  milk  above 
or  below  the  genetic  base  that  the  daughters  of 
a bull  will  produce  on  average  due  to  the  genes 
they  receive.  As  indicated  in  table  34,  the  pre- 
dicted difference  for  milk  yield  transferred  via 
the  bull  shows  an  improvement  from  122  to  908 
lb  for  active  AI  bulls  in  the  United  States  over 
the  past  13  years. 

This  impressive  improvement  still  lags  behind 
what  is  theoretically  possible.  A hypothetical 
breeding  program  could  result  in  an  expected 
yearly  gain  of  220  lb  of  milk  per  cow,  using  AI; 
and  the  biological  limits  to  this  rate  of  gain  are 
not  known.  In  practice,  the  observed  genetic 
trend  in  the  U.S.  national  dairy  herd  is  about 
100  lb— 70  lb  from  the  PDs  of  hulls  plus  30  lb  or 
so  from  the  female,  most  of  which  is  actually 
carryover  effect  from  the  previous  use  of  supe- 
rior bulls. 

AI  organizations,  many  of  which  are  coop- 
eratively owned  by  dairymen,  have  not  rigor- 
ously applied  the  principles  of  AI.  Their  efforts 
have  been  limited  by  reluctance  to  break  with 
traditional  selection  practices,  financial  con- 
straints for  proper  testing  of  young  bulls  to  pro- 

Table  34.— Predicted  Difference  (PD) 
of  Milk  Yield  of  Active  AI  Bulls 


Year TO  milk  (lb) 


1967  122 

1968  198 

1969  205 

1970  276 

1971  301 

1972  346 

1973  348 

1974  336 

1975  425 

1976  501 

1977  558 

1978  748 

1979  908 


SOURCE:  Animal  Improvement  Programs  Laboratory,  Animal  Science  Institute, 
Beltsville  Agricultural  Research  Center,  USDA. 


duce  sires  of  cows,  and  too  much  emphasis  on 
nonproductive  traits  of  ciuestionahle  economic 
value.  The  progress  that  has  been  made  has  re- 
sulted from  the  increased  use  of  AI,  the  a\ail- 
ability  of  data  through  NCDlllP,  and  the  ac'tual 
use  of  reliable  genetic  exaluations.  If  an\'  of 
these  three  factors  had  been  missing,  far  less 
improvement  would  ha\  e occuri'ed. 

Semen  Storage.— It  is  doubtful  that  major 
technological  changes  in  processing  semen  w ill 
occur.  However,  since  the  I'ate  of  conception  is 
as  important  as  the  genetic  merit  of  a sir(>  to  th(> 
economy  of  a dairy  enterprise?,  more  attention 
will  be  given  to  selecting  sires  of  high  fei'tility. 
Progress  should  he  made  in  hanking  seiiu'ii  by 
AI  studs  as  a hedge  against  costs  of  inflation.  In 
the  future,  some  of  the;  ine-re'ase'el  e'e)sts  e>f  he)us- 
ing  and  feeding  hulls  will  preehahly  he*  e)ffset  by 
semen  hanking  and  earlie*r  e'limination  of  manv 
bulls. 

Sexed  Semen.— Se;xing  of  se*me*n  te)  pre)eluce* 
heifer  cakes  (fe)i'  dairvme’ii)  e)r  hull  e ak  e's  (for 
AI  organizatieins)  has  l)e;e*n  atte*mpteel  \\ithe)ut 
success  for  many  ye;ars. 

Perfect  determination  of  the*  se*\  of  proge*ny 
could  practically  elouhle*  sele*ction  inte'usity  in 
two  ways— with  elams  to  |)re)eluce*  hulls  for  te*st- 
ing  in  AI  and  dams  te>  proeluce  re*plae  e*ments.  It 
sexed  semen  is  use;el  with  .in  AI  plan,  the*  thi’o- 
retical  impi'ei\eme*nl  in  milk  yie*lel  woulel  he*  ;t:t 
Ih  per  year’,  with  2'A  Ih  elue*  to  se*le*ction  of  elams 
for  replacements. 

The  \ alue;  e)f  this  aelelitional  amount  pe*r  year 
may  ne)t  seem  gre*at  for  any  ineli\  ielual  cow  , hut 
when  it  is  multiplie*el  by  a national  h(*rd  ol  7 
millieen  cenvs  using  ,\l  anel  is  accuimil<ile*el  lor  10 
years,  the;  eceeneimie'  \alu(*.  at  ,S()  10  Ih.  is  .ihout 
$1.1  hilliem— an  a\  e*rage*  ol  $ I It)  million  per  \ ear 
and  $231  millie)n  eluring  the*  IDth  u*aiv  I he  cost 
e)f  sexing  seme*n  is  not  know  n.  since*  no  one  h.is 
successfully  de)ne*  it.  II  a wa\  is  lounef  the*  cost 
weuild  ha\e;  to  he*  unele*r  $10  per  hi'ceding  unit 
teir  the  pi'ejceelure*  to  he*  e*e  e)ne)mie-.il 

Embryo  1’raiisfrr.— I he*  transle*i  eil  li*i 
tilized  eggs  freem  a e eiw  tei  eihtain  |)reige*iw  h.is 
been  ae;ce)mplishe*el  w ith  gi  e*al  sue  e e*ss  Most 
transle*i\s  h;i\  e*  in\  eik  e*el  popular  eti'  e*\eilie  hi  l•l•el 


Ch.  9— Advances  in  Reproductive  Biology  and  Their  Effects  on  Animal  Improvement  • 185 


in^  animals  wilh  littln  rn^ard  lor  g(MuMic  poU’ii- 
tial. 

Kmhi'u)  transtor  ma\  pay  tor  ilsell  in 

ItM'iiis  ol  milk  proiluc'tion  ot  tht*  animals  pro- 
(liK'tnl  exc'opt  iiulirorlK  through  hulls.  Kalhor,  it 
is  used  mostly  to  produce  outstanding  row  s tor 
sale.  Other  eommereial  a{)pIieations  tor  eattle 
inelude  ohtaining  progein  trom  otherwise  in- 
tei'tile  rows,  exporting  t'liihi  xos  instead  of  ani- 
mals. and  tt'sting  toi’  reeessi\  (‘  genetic  traits. 

Kmhryo  transfer  progeny  must  he  worth 
each  to  justity  tlu>  costs  and  risks,  .\hout 
SI. 500  of  this  represiMits  costs  due  to  emhrxo 
transfer  and  SI. 000  the  costs  of  proilucing 
cakes  normally.  If  genetic  gain  from  emhryo 
transfer  comes  onl\  from  dam  paths,  the  e.\- 
[)ected  gain  ox  er  M alone  is  70  Ih  \ r.  K.xtra  gain 
at  SO. 05  II)  ahox  e feed  cost  would  hax  e to  ac- 
cumulate for  79  years  before  added  gain  w ould 
equal  exen  a S300  embryo  transfer  cost  per 
[)iegnancx.  If  less  semen  is  neetled  lalloxxing 
more  intensix  e hull  selection),  the  e.\j)ected  gain 
of  129  Ihyr  must  accumulate  for  40  years  to 
balance  an  emhiyo  transfer  cost  of  S300  per 
pregnancy. 

Emhrxo  transfer  and  perfect  se.xing  of  semen 
XX  ould  combine  to  im[)i  ox  e genetic  gain  (in  milk 
production)  slightlx.  The  use  of  less  semen 
might  be  possible  through  application  of  in  x itro 
fertilization.  Hoxxexer,  feasibility  based  on 
genetic  gain  xxould  still  require  holding  all  costs 
doxx  n to  around  S50  to  S90  per  conception.  The 
general  conclusion  is  that  costs  of  emhi  yo  trans- 
fer must  he  greatly  reduced  to  he  economically 
feasible  if  only  genetic  gain  is  considered. 

Estrus  Synchronization.— The  ax  ailability 
of  an  effectixe  estrus  synchronization  method 
XX  ould  prox  ide  strong  impetus  for  increased  use 
of  .AI  and  embryo  transfer  in  dairy  cattle.  The 
detection  of  estrus  is  an  e.xpensix  e operation;  ef- 
fectix  e control  of  estrus  cycling  also  requires  in- 
tensixe  management,  adequate  handling  facil- 
ities, and  close  cooperation  betxxeen  the  pro- 
ducer, x eterinarian,  and  .AI  technician. 

Summary.— 

• Proper  application  of  progeny  testing  xvith 
selection  and  AI  can  increase  the  genetic 


gain  for  milk  yield  more  than  Ixxo  times 
fast(M'  than  is  occurring  today.  Improxed 
exaluation  of  coxxs,  pro|)er  economic  em- 
phasis on  other  traits,  and  strict  adherence 
to  .seUu’tion  stanilards  are  the  keys.  Bio- 
logical limitations  to  this  rate  of  genetic  im- 
proxement  cannot  he  anticipated  in  the 
foreseeable  future. 

• AI  of  dairy  cattle,  xxith  the  present  intensi- 
ty of  sire  .selection,  should  increase  the  net 
xxorth  or  |)rofit  of  animals  (increased  x'alue 
minus  extra  costs  of  the  -A I pi'ogram)  about 
SlO.OO  head  per  year.  By  1990,  « million 
daily  coxxs  in  ,AI  programs  xxould  he  xxorth 
about  S800  million  (8  X i()«  X $i()  X 10 
years)  more  at  current  market  pi'ices  as  a 
result  of  continued  u.se  of  AI. 

• Se.xing  of  semen  xxhen  used  xxith  A I may 
pax  for  it.self  if  the  cost  per  breeding  unit 
can  h('  kept  biMxx  een  $10  and  $20. 

• Emhryo  transfer  is  unlikely  to  pay  for  itself 
genetically  unless  the  cost  is  reduced  to  be- 
txxeen $50  and  $90  i)er  conception.  Hoxv- 
ex  er,  des|)ite  its  high  costs,  it  is  used  to  pro- 
duce animals  of  e.xceplionally  high  x^alue. 
(See  app.  ll-(]  for  an  exjtlanation  of  reasons 
other  than  genetics  xvhy  embryo  transfer  is 
used.) 

• Estrus  synchronization  is  noxx'  ax  ailable  for 
use  xxith  heifers,  and  should  increase  the 
use  of  ,AI  and  consequently  the  genetic  im- 
prox  ement  of  dairy  cattle. 

• ,A  secondary  benefit  of  all  technologies  is 
the  increased  number  of  skilled  persons 
xx  ho  can  prox  ide  technical  skills  as  well  as 
educate  dairymen  in  all  areas.  Also,  a 
unique  pool  of  reproductive  and  genetic 
data  has  been  accumulated. 

BEEF  CATTLE 

There  is  no  single  trait  of  overriding  im- 
portance (like  milk  production  in  dairy  cows)  to 
emphasize  in  the  genetic  improvement  of  beef 
cattle,  the  rate  of  growth  is  a possibility.*  It  is 
also  difficult  to  select  for  several  traits  at  once, 


'Beef  and  dairy  cattle  are  usually  different  breeds  in  the  United 
States.  In  the  literature  and  in  research  they  are  often  referred  to 
as  different  species.  In  other  countries,  notably  in  VX’estern  Europe 
and  in  Japan,  so-called  "dual  purpose"  cattle  are  used  to  produce 
both  beef  and  milk.  In  the  United  States,  old  dairy  cows  usually  be- 
come hamburger. 


186  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


especially  when  some  are  incompatible— e.g.,  it 
is  desirable  to  produce  large  animals  to  sell,  but 
undesirable  to  have  to  feed  large  mothers  to 
produce  them.  There  are  also  other  complica- 
tions. Growth  rate  has  two  genetic  components^ 
for  which  one  can  select— the  maternal  con- 
tribution (primarily  milk  production)  and  the 
calf’s  own  growth  potential.  Other  traits  of  in- 
terest are  efficiency  of  growth,  carcass  quality 
traits  (such  as  tenderness),  calving  ease,  and 
reproductive  traits,  such  as  conception  rate  to 
first  service  with  AI. 

Genetic  improvement  programs  for  beef  have 
two  major  advantages  over  those  for  dairy  cat- 
tle traits  such  as  growth  rate  and  carcass  quality 
can  he  measured  in  both  sexes  (whereas  one 
cannot  measure  the  milk  production  of  bulls); 
and  the  traits  are  moi’e  heritable  than  milk 
production. 

Artificial  Insemination. — Between  3 and 
5 percent  of  the  U.S.  beef  herd  is  artificially  in- 
seminated each  year.  This  low  rate  is  due  to  sev- 
eral factors,  including  management  techniques 
(range  v.  confined  housing),  availability  of  re- 
lated technologies  (especially,  until  recently, 
estrus  synchronization),  and  the  conflicting  ob- 
jectives of  the  indix'idual  breeders,  ranchers, 
and  breed  associations. 

Because  little  is  known  about  the  effective- 
ness of  AI  in  spreading  specific  genes  through- 
out the  Nation’s  beef  herds,  analysts  have  con- 
centrated on  their  reproductive  performance. 
Calf  losses  are  heavy  throughout  the  Nation. 
The  calf  crop— the  number  of  calves  alive  at 
weaning  as  a fraction  of  total  number  of  females 
exposed  to  breeding  each  year— is  estimated  to 
be  between  65  and  81  percent.  To  put  these 
data  in  perspective,  USDA^®  has  estimated  that  a 
5-percent  increase  in  the  national  calf  crop 
would  yield  a savings  of  $558  million  per  year  in 
the  supply  of  U.S. -grown  beef.  Techniques  now 
available  can  produce  such  an  increase  when 
they  are  integrated  into  an  adequate  manage- 
ment program. 


'"II. S.  Ueparlnient  of  /Xgriculture,  Agricultural  Resea]‘ch  Ser\'- 
ice,  "Beef  I’i'odiiction,"  ARS  National  Re.search  Program  Report  No. 
203H0  (VV'a.shington,  D.C.:  flSD/\,  October  1976). 


The  standardized  measure  of  weaning  weight 
in  beef  cattle  is  the  weight  at  205  days,  adjusted 
for  sex  of  calf  and  age  of  dam.  In  a recent  study 
in  West  Virginia— the  Allegheny  Highlands  Proj- 
ect-calf weights  ha\  e averaged  an  increase  of 
10  lb  per  year  of  participation  in  the  pi'oject,  \ ia 
AI  and  crossbreeding.  Estimates  of  increased 
value  of  calves  statewide,  should  the  same  tests 
and  AI  program  be  expanded,  add  up  to  $3.6 
million  per  year  when  calf  prices  a\  erage  $50 
per  hundredweight. Rapid  adoption  of  ,\l 
could  bring  about  this  kind  of  increase  in  as  lit- 
tle as  40  to  48  months. 

The  costs  and  returns  of  ,\l  \ arv  from  farm  to 
farm  and  with  the  numhei'  of  cattle  in  ('strus.  In 
general,  it  becomes  more  \ aluahle  w ith  smallei- 
herds,  more  cows  in  estrus,  higiK'i'  conce|)tion 
rates,  and  better  hulls.  Eoi-  purc'hred  herds, 
even  larger  benefits  have  been  estimated — e.g., 
in  a 1969  study,  the  estimated  inci'ease  in  \alue 
per  calf  when  AI  was  used  was  $30.02  on  pure- 
bred ranches  compared  to  $3.31  on  commercial 
ranches  in  Wyoming.^® 

A major  secondary,  or  indirect,  hi'iicMit  of  the 
use  of  AI  is  feed  sa\'ed  for  other  uses.  It  has 
greatly  reduced  the  numln'r  of  sires  lu'cessaiy 
for  stud  serxice  and,  thi’ough  radically  im- 
proxed  milk  prodiK'tion,  the  inimhei-  of  females 
as  xvell.  rhese  nnluced  re(|uirements  together 
are  e(|uix  alent  to  more'  than  1 billion  hu  of  corn 
and  other  concentrates.  Ibis  situation  xxill  he 
further  enhanced  as  beef  cattle  AI  expands 

Synchronization  of  ilstriis. — Diflerences 
in  the  rates  of  application  ol  AI  hetxxeen  heel 
and  dairy  herds  can  Ix'  explained  part  lx  by  the 
differing  managenuMil  .systems  loi-  the  Ixxo 
tyjies  of  classes  of  cattle.  Dairx  herds  arc  kept 
close  to  the  barai  for'  milking  and  are  accus 
tomed  to  being  approar  hed  In  humans  In  con- 
trxrst,  beef  her'ds  may  numhei’  a fexx  thmrsand 
head  on  100, ()()()  acr’es  ol  ar  id  paslurr*  land  I he 
detection  of  estrus  under  Ihesi*  conditions  is 
difficult. 


"'R.  S.  liilklM'.  XI  R I .IllM-ll  P I 1 C\M\  .mil  I K 111  kl  1|I  X 
Pmgr;im  Report  on  the  Xlleghen\  llighl.inil-.  I’ii‘|eil  '\|,  i,;.e 
tou  ii,  XX  . X it.:  XX Cst  X irgini.i  I im  eiMt\  l.inii.ii  \ is  ■ einbi  i | i:  o 
^"D  M Sle\  en.s  ;iiul  I Xtolir  Xrtiliii.il  lii-i-iiii  i.ili.'o  >1  ll.m.:' 
Cattle  in  XX  voming:  An  I.eonomic  Xii.ib  sis  XX  muiiiuk  \|  i . nlin,  , 
lAperiment  Station  Bulletin  \o  I'M,  I'M.'t 


Ch.  9 — Advances  in  Reproductive  Biology  and  Their  Effects  on  Animal  Improvement  • 187 


It  has  piviliftfil  tlial  tlu>  a\ ailahility  ot 

prosta^laiuliii  a^'ents  tor  regulating  estrus  eoulcl 
increase  tlie  numlier  ol  heel  eal\es  horn  trom 
superior  hulls  hy  ID  times,  and  that  perhaps  20 
[lereent  ol  the  I S.  heel  eow  herd  coukl  reeei\  e 
at  least  one  insemination  aiiilii  ally  hy  1090.-'  It 
this  lead  to  a ">0-lh  inerease  in  weight  tor  10  [)er- 
eent  ot  the  (\il\t*s  hoi'ii.  it  shoulil  ht*  worth  SI  14 
million  to  St 22  million  each  \ear.  assuming  80 
or  8a  [lereent  net  call  ei’op  and  SOO  per  hun- 
ilreilw  eight 

The  implementation  of  reeiMith  ilexeloped 
estrus  s\  nehronization  teehnologx  might  in- 
ereast'  the  numht*r  ot  heel  eow  s hred  artit'ieially 
In  4,000.t)00  in  the  I'nitetl  States.  Such  a pro- 
gram shoulil  he  sueeesstui  in  athaneing  the 
caking  date  In  one  week  (hy  decreasing  the 
cak  ing  interx  all.  and  in  increasing  the  i|uality  of 
the  cak  es  produced.  These  new  cak  es  could  he 
vvoi'th  about  SI 00  million  annually,  less  about 
SaO  million  ilue  to  e.vtra  costs  associated  with 
the  s\  nehronization  j)rogram. 

Sex  Control. — Se.\  control  would  have  a 
dramatic  effect  on  the  beef  industry.  In  1971,  it 
was  projected  that  In  1980  sex  control  could 
ha\  e an  annual  potential  benefit  of  S200  million 
based  on  10  million  female  cakes  being  re- 
placed In  male  calves  produced  through  the 
sexing  of  semen.--  .At  the  time  of  the  prediction, 
the  market  \ alue  for  steers  was  about  S20  more 
than  for  heifers.  (Steers  w ean  hea\  ier  and  gain 
more  efficiently.)  Now  the  margin  is  much 
greater— approximately  S50.  This  potential 
method  of  biological  control  is  more  attractixe 
than  the  use  of  additixes  like  steroids  or  im- 
plants because  of  the  possible  hazards  associ- 
ated XX  ith  them  that  preclude  their  use. 

Embryo  Transfer. — The  possibilities  for 
genetic  improxement  in  beef  cattle  using  em- 
bryo transfer  haxe  been  analyzed.  It  appears 
that  embryo  transfer  programs  can  be  dexel- 
oped  to  increase  the  rate  of  genetic  progress  for 


-'H.  D.  Hal's.  "Potential  Impact  of  Prostaglandin  on  Prospects  for 
food  From  Dairy  Cattle."  Proc.  Luialyse  Symposium,  J.  XX'.  Lauder- 
dale and  J.  H.  Sokolowski  leds.)  (Kalamazoo.  Mich.:  Upjohn,  1979), 
pp.  9-14. 

“R  H.  Foote  and  P.  Miller.  XX  hat  Might  Se.x  Ratio  Control  Mean 
in  the  .Animal  XX  orld."  Symposium,  Am.  Soc.  of  Animal  Science, 
1971.  pp.  1-10. 


groxx  th  rate:  but  the  programs  are  much  too  ex- 
pensixe  to  he  used  oxer  the  entire  population. 
One  problem  is  that  the  economic  xalue  of  the 
product  of  a beef  coxx  is  around  2o  percent  (or 
exen  less)  of  that  of  a dairy  coxx'.  Nexei'theless, 
in  populations  in  xxhich  ,\l  is  usetl,  embryo 
transfer  xxas  found  to  he  useful  for  obtaining 
more  hulls  from  (op  coxvs.  The  females  pro- 
duced hy  emhi’xo  transfer  xxould  he  xxorth  mar- 
ginally more  than  females  produced  conxen- 
tionally,  hut  the  costs  and  influence  of  males 
could  spread  oxer  the  population  through  the 
use  of  AT  The  extent  of  this  use  of  embryo 
transfer  xxould  be  xery  small;  only  a fexv  hun- 
dred hulls  xxoultl  he  produced  per  year  for  x ery 
large  populations,  and  oxer  99  percent  of  the 
population  xvould  reproduce  conxentionally. 
Iloxxexer,  such  programs  could  haxe  consider- 
able economic  benefit.  Ciive.  must  be  taken  to 
minimize  increased  inbreeding  of  the  popula- 
tion XX  ith  such  a breeding  scheme. 

Su  miliary.— 

• A1  could  substantially  improx  e economical- 
ly important  traits  in  beef  herds.  Hoxvever, 
because  of  the  dixersitv  of  traits  consid- 
ered important  by  different  breed  groups 
and  tbe  lack  of  a national  beef  testing  and 
recording  system  comparable  to  NCDHIP, 
economic  estimates  of  its  value  have  not 
been  dex  eloped. 

• A sexing  technology  to  produce  mostly 
males  (they  groxv  faster  than  heifers)  could 
be  of  enormous  potential  benefit  to  the 
beef  industry.  Hoxvever,  no  successful 
technique  yet  exists. 

• Estrus  cycle  regulation  could  lead  to  a sub- 
stantial increase  in  the  number  of  beef  cat- 
tle in  A1  programs.  The  net  benefit  of  this 
technology,  coupled  with  AI,  may  be  as 
high  as  S50  million  per  year.  Similarly,  the 
availability  of  reliable  progeny  records 
xvould  add  to  the  beneficial  impact  of  AI  in 
beef  and  xvould  probably  contribute  sig- 
nificantly to  its  use  in  beef  cattle. 

OTHER  SPECIES 

Swine. — Much  progress  has  been  made  in 
improving  the  overall  biological  efficiency  of 
pork  production  in  the  United  States.  Improved 


188  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


growth  rales,  feed  efficiencies,  carcass  merit, 
and  litter  sizes  have  helped  keep  pork  prices 
down  and  improve  its  quality  in  the  Nation’s 
markets.  Pork  today  is  leaner  and  contains  more 
high-quality  protein  calories  than  it  was  just  a 
few  decades  ago. 

A1  in  swine  production  could  expand,  al- 
though it  will  be  limited  by  the  relatively  poor 
ability  of  swine  sperm  to  withstand  freezing 
and  by  the  problem  of  detecting  estrus.  It  will 
be  encouraged  by  the  strong  trend  toward  con- 
finement housing  and  integration  of  all  phases 
of  hog  production.  The  industry— especially  the 
individual,  family-farm  type  units— would  bene- 
fit by  the  establishment  of  a progeny  testing 
scheme  to  identify  superior  boars.  Publicly 
available  information  on  genetic  merit  would 
decrease  dependence  on  a few  corporate  breed- 
ing organizations. 

Embryo  transfer  in  swine  will  be  strictly 
limited  by  difficulties  in  developing  nonsurgical 
methods  of  recovery  and  transfer,  and  by  the 
low  economic  value  per  animal  in  comparison  to 
cattle  and  horses.  However,  embryo  transfer  is 
useful  in  introducing  new  genetic  material  into 
breeding  herds  of  specific  pathogen-free  swine 
and  in  transporting  genetic  material  to  various 
regions  of  the  world. 

Sheep. — The  processes  of  selection  and  of 
crossing  specific  strains,  which  have  been  so  ef- 
fective in  poultry  and  hogs,  have  been  virtually 
ignored  in  sheep.  Selection  of  replacement  ewes 
from  the  fastest  growing  ewe  lambs  born  as 
twins  and  the  use  of  flushing  to  increase  ovula- 
tion rates  have  led  to  annual  increases  of  1.8 
percent  in  lambing;  in  one  test  tbe  market 
weight  of  lambs  was  increased  by  1/lb/yr  of 
cooperation.^® 

Synchronization  of  estrus  in  ewes  can  be 
achieved  with  prostaglandin  and  many  differ- 
ent progestogens.  The  technique  is  used  exten- 
sively in  many  countries,  but  no  products  for 
this  purpose  are  currently  marketed  in  the 
United  States. 

AI  rates  abroad  sometimes  approach  100  per- 
cent. However,  AI  will  not  be  used  widely  on 


K.  Inskeep,  personal  communication,  1980. 


sheep  in  the  United  States  until  systems  for  per- 
formance and  progeny  testing  are  implemented 
that  will  track  the  number  of  lambs  born  and 
their  growth  rate,  and  until  routine  freezing  of 
raw  semen  is  achieved. 

Goats. — The  research  performed  on  goats  is 
largely  designed  for  application  to  other  ani- 
mals. However,  interest  in  goats  in  the  United 
States  and  the  demand  for  their  products 
through  the  world  is  increasing. 

NCDHIP  has  just  started  providing  sire  e\al- 
uations  to  goat  breeders.  These  data,  along  w ith 
artifical  insemination,  should  increase  milk  pro- 
duction. The  genetic  data  might  he  of  particular 
usefulness  in  the  less  de\eloped  countries 
where  most  goat  raising  occurs.  Ureater  use  of 
all  reproductive  technologies  on  valuable  Ango- 
ra goats  might  be  expected. 

Other  technologies 

Tbe  use  of  any  reliable  twinning  or  s('.\  s(*l('c- 
tion  technologies  will  he  limited  until  such  |)ro- 
cedures  can  he  made  simple,  fast,  ini’xpensiv c. 
and  innocuous.  No  widesi)i'ead  u.se  of  thes(> 
technologies  should  he  expcH’ted  \\  ithin  tlu'  next 
decade. 

The  more  esoteric  techni(|ues  Ibi-  manipu- 
lating sex  cells  or  the  germplasm  its('lf  w ill  ha\ c 
no  impact  on  the  production  of  animals  or 
animal  products  within  the  next  20  years.  In 
vitro  manipulations,  including  cloning,  ci'll  fu- 
sion, the  production  of  chimeras,  and  the  use  of 
rDNA  lechnic|Lies,  u ill  continue  to  he  of  inten.se 
interest.  However,  it  is  unlikely  that  they  will 
have  practical  effects  on  farm  production  in  the 
United  States  in  this  century.  I.aeh  teehni(|ue 
will  require  more  resc’ari'h  and  refinement  Un- 
til specific  geiKks  can  b(^  identified  and  locat('d. 
no  direct  gene  manipulation  will  be  pr.ictic.ible 
A polygenic  basis  for  most  liaits  of  importance 
can  be  expected  to  be  th(>  rule  rath(M'  th.in  the 
exception. 

Should  such  techniques  become  a\ailal)le. 
limited  use  for  producing  breeding  stock,  can  be 
expected.  Experience  with  eai'ly  users  of  ,\1  and 


“Ibid. 


Ch.  9 — Advances  in  Reproductive  Biology  and  Their  Effects  on  Animal  Improvement  • 189 


embryo  transfer  is  strong  evidence  for  the  pre- 
dicted use  of  the  technologies,  no  matter  what 
their  economic  justification.  (See  app.  11-C.) 

.A  major,  secondary  effect  of  animal  research 
in  reproductive  biologv'  is  increased  under- 
standing leading  to  the  possible  solution  of 
human  problems— e.g.,  the  concept,  efficacy, 
and  safety  of  the  original  contraceptive  pill  was 
developed  and  established  in  animals.  It  in- 
volves the  same  principle  as  estrous  cycle  reg- 
ulation discussed  above. 

\(U  \Cl  l,Tl  KK 

Aquaculture  is  the  cultivation  of  freshwater 
and  marine  species  (the  latter  is  often  referred 
to  as  mariculture).  W bile  fish  culture  is  about 
6,000  years  old,  scientific  understanding  of  its 
basic  principles  is  far  behind  that  of  agriculture. 
.Aquaculture  is  slowly  being  transformed  into  a 
modern  multidisciplinary  technology,  especially 
in  the  industrialized  countries.  Increasing 
awareness  of  human  nutritional  needs,  over- 
fishing of  natural  commercial  fisheries,  and  ris- 
ing worldwide  demand  for  fish  and  fish  prod- 
ucts are  trends  that  indicate  a growth  in  inter- 
est in  aquaculture  as  a means  to  meet  the  food 
needs  of  the  world’s  population. 

As  part  of  the  trend  toward  the  high  tech- 
nology and  dense  culturing  of  intensive  aqua- 
culture systems  in  the  industrialized  countries, 
problems  of  I'eproductive  control,  hatchery 
technologv,  feeds  technologv,  disease  control, 
and  systems  engineering  are  all  being  investi- 
gated. Reproductive  control  and  genetic  selec- 
tion are  important  because  most  commercial 
aquaculture  operations  must  now  depend  on 
wild  seedstocks.  \'ery  little  information  on  the 
animals  in  culture  is  av  ailable. 

V\  ith  all  three  of  the  aquaculture  genera  (fish, 
mollusks,  and  crustaceans),  selective  breeding 
programs  have  long  been  established,  healthy 
gene  pools  are  available,  and  advantageous  hy- 
bridizations have  been  developed.  In  fish  rais- 
ing, culture  systems  often  demand  sterile  hy- 
brids, especially  of  carp  and  tilapia.  Selective 
breeding  of  salmon  has  been  limited  by  political 
pressures.  V erv  little  work  has  been  conducted 
with  catfish,  the  largest  aquaculture  industry  in 
the  United  States.  The  use  of  frozen  sperm, 


w hich  has  been  successful,  should  increase  be- 
cause of  (be  sav  ings  in  transport  costs.  Although 
culture  systems  for  mollusks  are  fairly  well- 
dc'fined,  little  a|)plied  genetics  work  has  been 
done  with  these  po[)ular  marine  species.  Some 
success  has  been  reported  in  selection  for 
growth  rate  and  disease  resistance  of  the 
■ Xmei'ican  oyster,  and  selection  for  gi’ovvth  rate 
of  the  slow-growing  abalone  is  underway.  The 
crustaceans,  of  w hich  the  Louisiana  crayfish  is 
the  largest  and  most  viable  industry,  are  the 
least  undei’stood.  Successful  hybrids  of  lobsters 
bav  e been  dev  eloped. 

Aquaculture  suffers  from  an  insufficient  re- 
search base  on  the  species  of  interest.  However, 
growing  appreciation  of  and  demand  for  ma- 
rine species  should  result  in  increased  support 
for  basic  and  developmental  work  on  all  aspects 
of  control,  including  basic  reproductive  biology. 

POULTKY  BREEDING 

rhe  (|uantitative  breeding  practices  of  com- 
mercial breeders  have  changed  very  little  over 
the  last  30  years.  Highly  heritable  traits,  such 
as  growth  I’ate,  body  conformation,  and  egg 
weight,  ai’e  perpetuated  by  mass  selection  be- 
cause little  advantage  is  gained  from  hybrid 
vigor.  Low  heritable  traits  (egg  production,  fer- 
tility, and  disease  resistance)  are  perpetuated  by 
crossbreeding  and  identified  through  progeny 
and  family  testing. 

The  goals  of  the  industry  are  to  increase  egg 
production  of  the  layers— both  in  quality  and 
quantity— and,  with  broilers  and  turkeys,  to  im- 
prove growth  rate,  feed  efficiency,  and  yield,  as 
well  as  to  reduce  body  fat  and  the  incidence  of 
defects. 

The  technologies  of  AI  and  semen  preser- 
vation have  accelerated  the  advances  made 
through  quantitative  breeding  technology.  AI  is 
widely  used  in  commercial  turkey  breeding  be- 
cause of  the  inability  of  modern  strains  to  mate. 
It  makes  breeding  tests  more  efficient,  steps  up 
selection  pressure  on  the  male  line,  reduces  the 
number  of  necessary  breeder  males,  and  in- 
creases the  number  of  females  that  may  be 
mated  to  one  male.  Semen  diluents  were  intro- 
duced to  the  turkey  industry  about  10  years  ago 
to  lower  the  cost  of  AI.  Currently,  a little  over 


190  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


lialt  ot  the  turkeys  are  inseminated  with  diluted 
semen. 

lh'eser\ation  of  poultry  semen  by  freezing  is 
now  pi'acticed  by  several  primary  breeders.  Al- 
though freezing  chicken  semen  causes  it  to  lose 
some  potency,  the  practice  allows  increased  ge- 
netic advancement  and  the  distribution  of  ge- 
netic material  worldwide. 

The  amount  of  genetic  variation  available  for 
breeding  stock  is  not  expected  to  diminish  in  the 
near  future.  C^eilings  for  certain  traits  will  even- 
tually he  reached,  but  certainly  not  in  the 
1980's.  Advances  in  breeding  laying  chickens 
will  be  less  dramatic  than  in  the  past,  but  efforts 
will  continue  to  develop  new  genetic  lines  and 
to  improve  reserve  lines  and  crosses  to  meet  fu- 
ture needs. 

The  growth  rate  of  broilers  will  continue  to 
increase  at  4 percent  a year,  which  suggests 
that  birds  will  be  reaching  4.4  lb  in  5 weeks  by 
the  1990’s.  Breeding  for  stress  resistance  will  be 
increasingly  important,  not  only  because  of  the 
increased  use  of  intensive  production  systems, 
hut  also  to  meet  the  physiological  stresses  re- 
sulting from  faster  growth  and  greater  weight. 


AI  will  assume  increasing  impoi'tance.  Recent 
advances  in  procedures  for  long-term  freezing 
of  chicken  semen  will  allow  breeders  to  extend 
the  use  of  outstanding  sires.  The  sale  of  frozen 
seman  may  eventually  substitute,  in  part,  for 
the  sale  of  breeder  males. 

Dwarf  broiler  breedei's  will  also  assume  in- 
creasing importance  o\er  the  new  few  years. 
The  dwarf  breeder  female  is  approximately  25- 
percent  smaller  than  the  standard  female,  and 
even  though  the  dwarf’s  egg  is  smaller  and  the 
progeny’s  grou'th  rate  slightly  less  than  that  of 
the  standard  broiler,  the  lower  cost  of  produc- 
ing broiler  chicks  from  the  dwarf  breeder  more 
than  offsets  the  slight  loss  in  their  grow  th  rate. 
Dwarf  layers  and  the  dwarf  brecnler  Ikmis  could 
reduce  production  costs  by  20  peix'ent  and  2 
percent,  respecti\ely. 

There  is  some  inter(;st  among  poultry  breed- 
ers in  cloning,  gene  transfer,  and  sex  conti'ol 
but  progress  toward  succ(?s.sful  tc'chnologies  is 
slow. 


Issue  and  Options  for  Agriculture — Animals 


ISSUE:  Should  the  United  States  in- 

crease support  for  programs  in 
applied  genetics  for  animals  and 
animal  products? 

Advocates  of  a strong  governmental  role  in 
support  of  agricultural  research  and  develop- 
ment (R&D)  have  traditionally  referred  to  the 
small  size  of  the  production  unit:  U.S.  farms  are 
too  small  to  support  R&D  activities.  Throughout 
this  century  a complicated  and  extensive  net- 
work of  Federal,  State,  and  local  agricultural 
support  agencies  has  been  developed  to  assist 
the  farmer  in  applying  the  new  knowledge  pro- 
duced by  research  institutions.  This  private/ 
public  sector  cooperative  network  has  pro- 
duced an  abundant  supply  of  food  and  fiber, 
sometimes  in  excess  of  domestic  demand.  Social- 
ly oriented  policies  have  been  adopted  to  soften 


the  impacts  of  new  technology  and  to  rescue  the 
marginally  efficient  farmer  from  bankruptcy. 

Current  projections  of  U.S.  and  world  popula- 
tion growth  show  incix'asing  d(>mand  lor  all 
food  products.  Other  piH'dictable  trends  with 
implications  for  agricultural  lt(Si.D,  include: 

• growth  in  inconu^  for  some  populations, 
which  will  probably  inci-ease  the  demand 
for  sources  of  meat  piotein; 

• increasing  compcMition  among  \arious 
sources  of  protein  for  the  consuiin*rs 
dollar; 

• increasing  awareness  of  nutrition  issiu’s 
among  U.S.  consumers: 

• increasing  com[)(Uition  foi'  prime  agricul- 
tural land  among  agricultural,  urban,  .md 
industrial  intcMcsts: 

• increasing  demand  for  I .S  loud  and  tibcr 


Ch.  9 — Advances  in  Reproductive  Biology  and  Their  Effects  on  Animal  Improvement  • 191 


[)i'()diu'ls  tVom  abroatl,  U’acliiig  to  o[){)or- 
timities  for  increased  profits  for  siuTossfiil 
proiliicei's:  and 

• incroasing  demancis  on  agricultural  |)rod- 
ucts  for  pioduction  of  cnci  gx'. 

OPTIOi\S: 

Governmental  fmrticipation  in,  and fiiiuiin^  of, 
programs  like  the  \'ational  Cooperative  Dairy 
Herd  Improvement  Program  (XCDHIP)  could 
he  increased.  The  efforts  of  the  Beef  Cattle 
Improvement  Federation  to  standardize  pro- 
cedures could  be  actively  supported,  and  a 
similar  information  system  for  swine  could  he 
established. 

rhe  fastest,  least  expensive  way  to  u[)gi'ade 
breeding  stock  in  the  I'niled  States  is  through 
effective  use  of  information.  Clompuler  technol- 
ogv',  along  with  a network  of  local  represent- 
atives for  data  collecting,  can  |)rov  ide  the  imli- 
V idual  farmer  or  breeder  w ith  accurate  infor- 
mation on  the  gei'mplasm  available,  so  that  he 
can  then  make  his  own  breeding  decisions.  In 
this  way,  the  Nation  can  take  adv  antage  of  pop- 
ulation genetics  atid  information  handling  capa- 
bilities to  upgrade  one  of  its  most  ini[)ortant 
forms  of  capital:  poultry  and  livestock.  Breed 
associations  and  lai'ge  ranchers  who  sell  the 
semen  from  their  prize  hulls  based  on  pedigrees 
rather  than  on  genetic  merit  mav  act  as  harriers 
to  the  effectiveness  of  such  an  objective  infor- 
mation system. 

The  benefits  of  such  programs  would  accrue 
both  to  L'.S.  consumers,  in  reduced  real  prices 
of  meat  and  animal  products,  and  to  producers 
who  participate  in  the  programs,  in  increased 
efficiency  of  production.  Consumers  spend  such 
a large  part  of  their  incomes  on  red  meat  that 
ev  ery  increase  in  efficiency  represents  millions 
of  dollars  saved.  Beef  producers  too,  should 
welcome  any  assistance  in  upgrading  their 
stocks.  The  price  of  semen  has  remained  rel- 
atively stable,  and  semen  from  bulls  rated 
highly  on  certain  economic  traits  costs  only  a 
few  dollars  more  than  that  from  average  bulls. 

Howev  er,  efficiency  of  production  is  not  the 
only  value  to  be  upheld  in  U.S.  agriculture— e.g., 
in  milk  production  complex  policies  have  been 


designed  to  maintain  constant  milk  supplies 
without  large  fluctuations  in  price. 

The  NCUHIP  model  program  for  dairy  cattle 
has  shown  that  an  effective  national  program 
retjuires  the  participation  by  the  varied  in- 
terests in  program  policymaking  in  an  extension 
network,  for  local  collection  and  validation  of 
data  and  for  education  and  of  expertise  in  data 
handling  and  analysis.  Also  important  is  a 
strong  lead(M'ship  I’ole  in  establishing  the  pi'o- 
gram.  This  option  implies  that  the  l*’(Hleral  Gov  - 
('rnment  would  play  such  a role  in  new  pro- 
grams and  e.\|)and  its  role  in  existing  ones. 

B.  Federal  funding  of  basic  research  in  total  ani- 
mal improvement  could  be  increased. 

'I'he  o|)tion,  in  contrast  with  option  A, 
assuiiK's  that  it  is  necessary  to  maintain  or  ex- 
pand basic  R&.l)  to  generate  new  knowledge 
that  can  he  applied  to  the  production  of  im- 
proved animals  and  animal  products. 

Information  presented  in  this  repoi't  supports 
the  conclusion  that  long-term  basic  research  on 
the  physiological  and  biochemical  events  in 
animal  development  results  in  increasing  the  ef- 
ficiency of  animal  production,  both  in  total 
animal  numbers  and  in  quality  of  product.  In- 
creased understanding  of  the  interrelationships 
among  various  systems— including  reproduc- 
tion, nutrition,  and  genetics— gradually  leads  to 
the  development  of  superior  animals  that  effi- 
ciently consume  food  not  palatable  to  humans 
and  are  resistant  to  disease. 

Earlier  studies  also  support  the  importance  of 
basic  research— e.g.,  the  National  Research 
Council  found  in  1977  that  “.  . . not  as  much  fun- 
damental research  on  animal  problems  has 
been  conducted  in  recent  years  ...  it  should 
receive  increased  funding. USDA  also  found, 
in  a review  of  various  conference  proceedings, 
congressional  hearings,  special  studies,  and 
other  published  materials  on  agricultural  R&D 
priorities,  strong  support  for  more  research  on 
the  basic  processes  that  contribute  to  reproduc- 
tion and  performance  traits  in  farm  animals: 


“,\alional  Research  C:oiincil,  World  Food  and  Niilrition  Study, 
The  Potential  Contributions  of  Research  (Washington,  O.  C:.  author. 
1977),  p.  97. 


192  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Sp(;cit’ic  livestock  research  areas  identified  as 
having  signficant  potential  for  increased  pro- 
duction both  in  the  United  States  and  develop- 
ing countries  include:  1)  control  of  reproductive 
and  respiratory  diseases,  2)  developing  geneti- 
cally superior  animals,  3)  improving  nutrition 
efficiency,  and  4)  increasing  the  reproductive 
performance  of  all  farm  animal  species.^® 

^'^11.  S.  Oopyrtment  of  Agi'iciillure,  Science  ynd  Kducalion  Ad- 
niinisli'ylion,  Agricullural  and  Food  Research  Issues  and  Priorities 
1\\  yshinglon,  D.C.:  author,  1978),  p.  ,\iii. 


Regardless  of  the  effectiveness  of  present 
population  control  programs  or  of  current 
trends  in  individual  decisions  about  family  size, 
the  output  of  the  Nation’s  agricultural  activities 
must  increase  over  the  next  decades  if  sufficient 
food  is  to  be  available  for  the  woi'ld’s  popula- 
tion. Basic  research  is  the  source  from  which 
new  applications  to  increase  productiv  ity  arise. 


Part  III 


I'i 


I I 

• i 


I 


Institutions  and  Society 

. I 


Chapter  10.  The  Question  of  Risk 197 

Chapter  1 1.  Regulation  of  Genetic  Engineering 211 

Chapter  12.  Patenting  Living  Organisms 237 

Chapter  13.  Genetics  and  Society 257 


i 

• 1 


iti; 

■i\y 


chapter  10 

The  Question  of  Risk 


t 


^^3 


chapter  10 


Page 

Introduction 197 

The  Initial  Fear  of  Harm 197 

Classification  of  Potential  Harm . 198 

Identification  of  Possible  Harm 200 

Estimates  of  Harm:  Risk 200 

The  Status  of  the  Current  Assessment  of 

Physical  Risk 201 

Perception  of  Risk 203 

Burden  of  Proof 203 

Other  Concerns 204 

Concerns  Raised  by  Industrial  Applications  . . . 204 
Concerns  Raised  by  the  Implications  of  the 
Recombinant  DNA  Controversy  for  General 

Microbiology 204 

Concerns  Raised  by  the  Implications  of  the 
Recombinant  DNA  for  Other  Genetic 
Manipulation 206 


Page 


Ethical  and  Moral  Concerns 207 

Conclusion 207 


Figures 

Figure  No.  Page 

35.  Flow  Chart  of  Possible  Consequences  of  Using 
Genetically  Engineered  Micro-Organisms  ...  199 

36.  Flow  Chart  to  Establish  Probability  of  Harm 

Caused  by  the  Escape  of  a Micro-Oi'ganism 
Carrying  Recombinant  DNA 201 

37.  Alternative  Methods  for  Transferring  DN.A  From 

One  Cell  to  Another 206 


chapter  10 

The  Question  of  Risk 


Introduction  

The  perception  that  the  genetic  manipulation 
of  micro-organisms  might  gi\e  rise  to  unfore- 
seen risks  is  not  new . The  originators  of  chem- 
ical mutagenesis  in  the  1940's  were  warned  that 
harmful  uncontrolled  mutations  might  he  in- 
duced hv  their  techni(|ues.  In  a letter  to  the 
Recombinant  D\.\  Ad\  isorv  Committee  (RAC)  of 
the  National  Institutes  of  Health  (N'lH)  in  Decem- 
ber of  1979,  a pioneer  in  genetic  transformation 
at  the  Rockefeller  l'ni\  ersity,  w rote:  . I did 

in  1950,  after  some  deliberation,  perform  the 
first  drug  resistance  DN'A  transformations,  and 
in  1964  and  1965  took  part  in  early  warnings 
against  indiscriminate  transformations’  that 
were  then  being  imagined.”’ 

‘Kollin  I).  Hotrhkis.s.  Hectimhinant  l)\A  Hesearch,  vol.  o,  .MM  pul)- 
lic-iition  .\o.  80-2131).  March  1980.  p 484 


The  initial  fear  of  harm 

For  the  purposes  of  this  discussion,  harm  (or 
injury)  is  defined  as  any  undesirable  conse- 
quence of  an  act.  Such  a broad  definition  is  w ar- 
ranted  by  the  broad  targets  for  hypothetical 
harm  that  genetic  manipulation  presents:  injury 
to  an  indix  idual’s  health,  to  animals,  to  the  en- 
vironment. 

The  inital  concern  inx  oh  ed  injury  to  human 
health.  Specifically,  it  was  feared  that  combin- 
ing the  Di\A  of  simian  \ irus  40,  or  S\'40,  with  an 
Escherichia  coli  plasmid  would  establish  a new 
route  for  the  dissemination  of  the  virus.  Al- 
though the  S\'40  is  harmless  to  the  monkeys 
from  which  it  is  obtained,  it  can  cause  cancer 
w'ben  injected  into  mice  and  hamsters.  And 
while  it  has  not  been  shown  to  cause  cancer  in 
humans,  it  does  cause  human  cells  to  behave 
like  cancer  cells  u'hen  they  are  grown  in  tissue 
culture.  W'hat  effect  such  viruses  might  have  if 
they  were  inserted  into  E.  coli,  a normal  in- 
habitant of  the  human  intestine,  w'as  unknown. 
This  uncertainty,  combined  with  an  intuitwe 


\'et  none  of  this  earlier  public  concern  led  to 
as  great  a controversy  as  has  research  with  re- 
combinant DNA  (I'DNA).  No  doubt  it  was  en- 
couraged because  scientists  themselves  raised 
questions  of  potential  hazard.  The  subsequent 
open  debates  among  the  scientists  strengthened 
the  public’s  perception  that  there  w^as  legitimate 
cause  for  concern.  This  has  led  to  a continuing 
attempt  to  define  the  potential  hazards  and  the 
chances  that  they  might  occur. 


judgment,  led  to  a concern  that  something 
might  go  wrong.  The  dangerous  scenario  went 
as  follows: 

• SV40  causes  cells  in  tissue  culture  to  be- 
have like  cancer  cells, 

• S\'40-carrying  E.  coli  might  be  injected  ac- 
cidently into  humans, 

• humans  would  be  exposed  to  SV40  in  their 
intestines,  and 

• an  epidemic  of  cancer  would  result. 

This  chain  of  connections,  while  loose,  was 
strong  enough  to  raise  questions  in  at  least  some 
people’s  minds. 

The  virus  SV'40  has  never  actually  been 
shown  to  cause  cancer  in  humans;  but  the  po- 
tential hazards  led  the  Committee  on  Recombi- 
nant DNA  Molecules  of  the  National  Academy  of 
Sciences  (NAS)  to  call  in  1974  for  a deferment  of 
any  experiments  that  attempted  to  join  the  DNA 
of  a cancer-causing  or  other  animal  virus  to  vec- 
tor DNA.  At  the  same  time,  other  experiments. 


197 


198  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


that  were  thought  to  have  a potential  for  harm 
—particularly  those  that  were  designed  to 
transfer  genes  for  potent  toxins  or  for  resist- 
ance to  antibiotics  into  bacteria  of  a different 
species— were  also  deferred.  Finally,  one  other 
type  of  experiment,  in  which  genes  from  higher 
organisms  might  have  been  combined  with  vec- 
tors, was  to  be  postponed.  The  fear  was  that  la- 
tent "cancer-causing  genes”  might  be  inadver- 
tently passed  on  to  E.  coli. 

Throughout  the  moratorium,  one  point  was 
certain:  no  evidence  existed  to  show  that  harm 
would  come  from  these  experiments.  But  it  was 
a possibility.  The  scientists  who  originally  raised 
questions  wrote  in  1975: ".  . . few,  if  any,  believe 
that  this  methodology  is  free  from  risk.”^  It  was 
recognized  at  that  time  that  ".  . . estimating  the 
risks  will  be  difficult  and  intuitive  at  first  but 
this  will  improve  as  we  acquire  additional 
knowledge.”^  Hence  two  principles  were  to  be 
followed:  containment  of  the  micro-organisms 
(see  table  35,  p.  213)  was  to  be  an  essential  part 
of  any  experiment;  and  the  level  of  containment 
was  to  match  the  estimated  risk.  These  prin- 
ciples were  incorporated  into  the  Guidelines  for 
Research  Involving  Recombinant  DNA  Mole- 
cules, promulgated  by  NIH  in  1976. 

But  the  original  fears  surrounding  rDNA  re- 
search progressed  beyond  concern  that  humans 
might  be  harmed.  Ecological  harm  to  plants,  ani- 
mals, and  the  inanimate  world  were  also  consid- 
ered. And  other  critics  noted  the  possibility  of 
moral  and  ethical  harm,  which  might  disrupt 
both  society’s  structure  and  its  system  of  values. 

Classification  of  potential 
physical  harm 

Some  combinations  of  DNA  may  be  harmful 
to  man  or  his  environment— e.g.,  if  an  entire 
DNA  copy  of  the  poliovirus  genetic  material  is 
combined  with  E.  coli  plasmid  DNA,  few  would 
argue  against  the  need  for  careful  handling  of 
this  material. 

For  practical  purposes,  the  potential  harm 
associated  with  various  micro-organisms  is 


^Recombinant  DNA  Research,  vol.  1,  DHEW  publication  No.  (NIH) 
76-1138,  August  1976,  p.  59. 

^Ibid. 


shown  in  figure  35.  Each  letter  (A  through  L) 
represents  the  consequence  of  a particular  com- 
bination of  events  and  micro-organisms.  For  ex- 
ample, the  letters: 

A, C  represent  the  intentional  release  of  micro- 

organisms known  to  be  harmful  to  the 
environment  or  to  man— e.g.,  in  biologi- 
cal warfare  or  terrorism. 

B, D  represent  the  inadvertent  release  of 

micro-organisms  known  to  be  harmful  to 
the  environment  or  to  man— e.g.,  in  acci- 
dents at  high-containment  facilities 
where  work  is  being  carried  out  with 
dangerous  micro-organisms. 

E, I  represent  the  intentional  release  of  micro- 

organisms thought  to  be  safe  hut  which 
prove  harmful— when  the  safety  of  orga- 
nisms has  been  misjudged. 

F, J  represent  the  intentional  release  of  micro- 

organisms which  prove  safe  as  expected— 
e.g.,  in  oil  recovery,  mining,  agriculture, 
and  pollution  control. 

H,L  represent  the  inadvertent  release  of 

micro-organisms  which  have  no  harmful 
consequences— e.g.,  in  ordinary  accidents 
with  harmless  micro-organisms. 

G, K  represent  the  inadvertent  release  of 

micro-organisms  thought  to  be  safe  hut 
which  prove  harmful— ihe  most  unlikc'ly 
possible  consecjuence,  because  both  an 
accident  must  occur  and  a misjudgnu'nt 
about  the  safety  must  ha\’e  heiMi  made. 

Discussions  of  physical  harm  have  rei'ogni/.ed 
the  possibility  of  intentional  misusi'  hut  ha\c 
minimized  its  likelihood.  Fhe  GoiniMition  on  the 
Prohibition  of  the  Dexelopment,  IModuclion, 
and  Stockpiling  of  Bacteriological  (Biological) 
and  Toxin  Weapons  and  on  th(>ir’  Destruction-' 
which  was  ratified  by  both  the  Senate  and  the 
President  in  1975,* *  states  that  the'  signatories 
will  "never  develo[i  . . . biological  agi'iits  or  tox- 
ins . . . that  have  no  justification  for  prophylac- 
tic, protective,  or  other  j)(>ac('ful  purposes.  " 
Such  a provision  clearly  includes  miero-oiga- 
nisms  carrying  rDNA  molecules  or  th('  toxins 

■’C:onvention  of  the  I’t-ohibiliou  ol  the  I )i-v elopmenl  l’it«lin  lion 
and  Stockpiling  of  llaclei'iological  IHiologicall  and  Iomii  \\ra(Min^ 
and  On  Their  Destruction.  Washington  l ondon.  .tiul  Xtosiou 
Apr.  10,  1972:  enteri-d  into  force  on  Mai  26  1 97.5  126  I s 1 '•.so 

*As  of  1980,  80  countiies  have  ralilied  the  lie.ilv  anolln-i  in 
have  signed  hut  not  ratified 


Ch.  10— The  Question  of  Risk  • 199 


Figure  35.— Flow  Chart  of  Possible  Consequences  of  Using  Genetically  Engineered  Micro-Organisms 


Micro-organism 


- Knctjm  ha^^dnis 


Suspected  safe 


For  environment 


mar 


For  environment 


For  man 


laterttion0  releae^ 


B.  Inadwteni  r« 


C.  tTTtenlioPiil  release  LJ  Intentional  release 


I—  p.  laedvertent  re^r^fs^ 


Intentional  release 


-E.  Proves  hazardous  -I.  Proves  hazardous 


- F.  Remains  safe 


J.  Remains  safe 


— Inadvertent  release 


Inadvertent  release 


SOURCE;  Office  of  Technology  Assessment. 


G.  Proves  hazardous 

K. 

Proves  hazardous 

H.  Remains  safe 

L. 

Remains  safe 

produced  by  them.  It  must  be  assumed  that 
those  ^\  ho  signed  did  so  in  good  faith. 

While  there  is  no  way  to  judge  the  likelihood 
of  dex  elopments  in  this  area,  the  problems  that 
would  accompany  any  attempt  to  use  pathogen- 
ic micro-organisms  in  warfare— difficulties  in 
controlling  spread,  protection  of  one’s  own 
troops  and  population— tend  to  discourage  the 
use  of  genetic  engineering  for  this  purpose.* 
Similarly,  the  danger  that  these  techniques 
might  be  used  by  terrorists  is  lessened  by  the 
scientific  sophistication  needed  to  construct  a 
more  virulent  organism  than  those  that  can 

•.Although  stockpiling  of  biological  warfare  agents  is  prohibited, 
research  into  new  agents  is  not. 


already  be  obtained— e.g.,  encephalitis  viruses 
or  toxin-producing  bacteria  like  C.  botulinum  or 
C.  tetani. 

Some  discussions  have  centered  around  the 
possibility  of  accidents  caused  by  a break  in  con- 
tainment. Construction  of  potentially  harmful 
micro-organisms  will  probably  continue  to  be 
prohibited  by  the  Guidelines;  exceptions  will  be 
made  only  under  the  most  extraordinary  cir- 
cumstances. To  date,  no  organism  known  to  be 
more  harmful  than  the  organism  serving  as  the 
source  of  DNA  has  been  constructed. 

However,  the  biggest  controversy  has  cen- 
tered around  unforeseen  harm — that  micro- 
organisms thought  safe  might  prove  harmful. 


200  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Discussion  of  this  kind  of  harm  is  hindered  by 
the  difficulty  not  only  of  quantifying  the  prob- 
ability of  an  occurrence  but  also  of  predicting 
the  type  of  damage  that  might  occur.  The  differ- 
ent types  of  damage  that  can  be  conjured  up  are 
limited  only  by  imagination.  The  scenarios  have 
included  epidemics  of  cancer,  the  spread  of  oil- 
eating bacteria,  the  uncontrolled  proliferation 
of  new  plant  life,  and  infection  with  hormone- 
producing  bacteria. 

The  risk  of  harm  refers  to  the  chance  of  harm 
actually  occurring.  In  the  present  controversy, 
it  has  been  difficult  to  distinguish  the  possible 
from  the  probable.  It  is,  for  instance,  possible 
that  an  individual  will  be  killed  by  a meteor  fall- 
ing to  the  ground,  but  it  is  not  probable.  Analog- 
ous situations  exist  in  genetic  engineering.  It  is 
in  this  analysis  that  debate  over  genetic  engi- 
neering has  some  special  elements:  the  uncer- 
tainty of  what  kind  of  harm  could  occur,  the  un- 
certainty about  the  magnitude  of  risk,  and  the 
problem  of  the  perception  of  risk. 

Identification  of  possible  harm 

The  first  step  in  estimating  risk  is  identifying 
the  potential  harm.  It  is  not  very  meaningful  to 
ask:  How  much  risk  does  rDNA  pose?  The  con- 
cept of  risk  takes  on  meaning  only  when  harm  is 
identified.  The  question  should  be:  What  is  the 
likelihood  that  rDNA  will  cause  a specific  dis- 
ease such  as  in  a single  individual  or  in  an  entire 
population?  The  magnitude  of  the  possible  harm 
is  incorporated  in  the  question  of  risk,  but  dif- 
fers in  the  two  cases.  A statement  about  the  risk 
of  death  to  one  person  is  different  than  one 
about  the  risk  of  death  to  a thousand.  The  right 
questions  must  be  asked  about  a specific  harm. 

Since  no  dangerous  accidents  are  known  to 
have  occurred,  their  types  remain  conjectural. 
Identifying  potential  harm  rests  on  intuition  and 
arguments  based  on  analogy.  Even  a so-called 
risk  experiment  is  an  approximation  of  subse- 
quent genetic  manipulations.  That  is  why  ex- 
perts disagree.  No  uncontestable  “scientific 
method”  dictates  which  analogy  is  useful  or  ac- 
ceptable. By  their  very  nature,  all  analogies 
share  some  characteristics  with  the  event  under 
consideration  but  differ  in  others.  The  goal  is  to 


discover  the  one  that  is  most  similar  and  to 
observe  it  often.  This  process  then  forms  the 
basis  for  extrapolation. 

For  example,  it  has  been  argued  that  ecologi- 
cal damage  can  be  caused  by  the  introduction  of 
plants,  animals,  and  micro-organisms  into  new 
environments.  Scores  of  examples  from  histoi’v 
support  this  conclusion.  I’he  introduction  to  the 
United  States  of  the  Brazilian  water  hyacinth  in 
the  late  19th  century  has  led  to  an  infestation  of 
the  Southern  waterways.  Unconti’olled  spread 
of  English  sparrows  originally  imported  to  con- 
trol insects  has  made  eradication  programs  nec- 
essary. Countless  other  examines  are  confirma- 
tion that  biological  organisms  may,  at  times, 
cause  ecological  damage  when  introduced  into  a 
new  environment.  Yet  there  is  no  agreement  on 
whether  such  analogies  are  particularly  reltv 
vant  to  assessing  potential  dangei's  from  genet- 
ically engineered  organisms.  It  could  he  ar- 
gued—e.g.,  that  a genetically  engintHM'tuI  orga- 
nism (carrying  less  than  1 pei’cent  new  genes)  is 
still  over  99  percent  the  same  as  the  original, 
and  is  therefore  not  analogous  to  lh(>  "totally 
new"  organism  introduced  into  an  (‘cosystem. 
Some  experts  emphasize?  the  difhM’ences  be- 
tween the  situations;  othei's  emphasize?  the?  simi- 
larities. 

Other  analejgies  ha\e?  he?e?n  raise'el.  Ne’w 
strains  of  influenza  \ irus  arise*  re'geilarly.  Some* 
can  cause  epielemics  he?e'ause*  the*  pe)|)ulalie)n, 
never  before  expe)seel  te)  them,  e'arrie*s  no  pro- 
tective antibodies.  \'e?t  e?an  this  analog_v  sugge*sl 
that  relatively  harmless  strains  of  E.  roli  might 
be  transformed  inte)  e?[)iele?mie?  |)athoge*ns?  I he*r(* 
is  disagreement,  anel  eie?hale*s  e-onlinue*  about 
what  "could  happen"  e>r  what  is  e*\(*n  logie'ally 
possible. 

Estimates  of  harm:  risk 

Assuming  that  agre?e?me*nt  has  he*e*n  reae  he*el 
on  the  possibility  of  a spe?e*ifie'  harm,  w hat  can  he 
done  to  ascei’tain  the?  probiibility'^  W hat  is  the* 
likelihood  that  elamage?  will  oe'cur? 

Damage  invariably  oe?e  urs  as  the*  result  ol  .i 
series  of  events,  e?ach  e)f  whie  h has  its  own  par 
ticular  chane;e  e)f  e)ce'uri  ing.  flow  charts  h.we 
been  prepare?el  te>  iele?ntifv  the*se*  ste*|)s.  \ t\pie-.il 


Ch.10 — The  Question  of  Risk  • 201 


analysis  cietermines  a probability  \ aluo  tor  eacb 
sli’[)— e.^..  in  ti^uro  :Ui  slop  II  tbn  [)i'()bability  ot 
es('a|)f  can  be  estimated  based  on  tbe  bistorical 
record  of  experiments  with  micro-  organisms. 
Depeiuling  on  tbe  degi'ee  ot  containment,  tbe 
j)i'obabilit\  \aries.  It  is  almost  certain  that 
expc'riments  on  an  op('n  beneb  top,  using  no 
precautions,  will  result  in  some  escape*  to  tbe 
surrouiuling  en\  ironment— a much  less  likely 
e\ent  in  maximum  containment  facilities.  (See 
table  3.1.) 

Two  points  sbould  bi*  noted,  first,  eacb  prob- 
ability can  be  minimized  by  appropriate  control 
measures.  Second,  tbe  probability  that  tbe  final 
e\  ent  w ill  occur  is  etiual  to  oi'  less  likeb  than  tbe 
least  likely  link  in  tbe  chain,  bec'ause  tbe  |)rob- 
al)ilities  must  be  multiplied  together,  if  the 
probabiliu  of  an\  single  step  is  zero,  tbe  prob- 
ability of  the  final  outcome  is  zei'o:  the  chain  of 
e\  ents  is  broken. 

THE  STATl  S OF  THE  CI  HKENT  .\SSES.SME,\T 
OF  PHYSICAL  RISK 

.\  successful  I'isk  assessment  sbould  pro\  ide 
information  about  tbe  likelihood  and  magnitude 
of  damage  that  might  occur  under  gi\en  cir- 
cumstances. It  is  clear  that  tbe  more  types  of 
damage  that  are  identified,  the  moi'e  risk  assess- 
ments must  be  carried  out. 


Figure  36.— Flow  Chart  to  Establish  Probability  of 
Harm  Caused  by  the  Escape  of  a Micro-Organism 
Carrying  Recombinant  DNA 


Event 


Probability 


I.  Inadvertent  incorporation  of  hazardous  gene 
into  micro-organism 


II.  Escape  of  micro-organism  into  environment 

f 

III.  Multiplication  of  micro-organism  and 
establishment  in  ecological  niche 

4 

IV.  Infection  of  man 

f 

V.  Production  of  factor  to  cause  disease 


P 


1 


^2 


P 


3 


P 


4 


P 


5 


NOTE:  Ps  will  always  be  smaller  than  any  of  the  other  probabilities. 


SOURCE;  Office  of  Technology  Assessment. 


.Although  the  original  charter  of  RAC  under- 
.scored  the  importance  of  a risk  assessment  pro- 
gram, it  was  not  until  f979  that  the  details  of  a 
formal  program  were  published.  For  5 years, 
risks  were  assessed  on  a case-bv-case  basis 
through:  1)  experiments  carried  out  under  con- 
tract from  iMIH,  2)  experiments  that  were  de- 
signed for  other  purposes  but  which  proved  to 
be  reUnant  to  tbe  c|uestion  of  risk,  and  3)  con- 
ferences at  which  findings  were  examined. 

From  tbe  start,  it  was  difficult  to  design  ex- 
periments that  could  supply  meaningful  infor- 
mation—e.g.,  bow  does  one  test  tbe  possibility 
that  "massive  ecological  disruptions  might 
occur?”  Or  that  a new  bacterium  with  harmful 
unforseen  characteristics  will  emerge?  Still 
some  experiments  were  proposed.  But  because 
tiiese  exfjeriments  bad  to  be  approximations  of 
tbe  actual  situation,  tbe  applicability  of  their 
findings  was  debated.  Here  too,  experts  could 
and  did  disagree— not  about  tbe  findings  them- 
seb  es,  but  about  their  interpretation. 

For  exani|)le,  in  an  important  experiment  de- 
signed to  test  a "worst  case  situation,”  a tumor 
\ irus  called  polyoma  w as  found  to  cause  no 
tumors  in  test  animals  when  incorporated  into 
E.  coli.^*  Since  just  a few  molecules  of  the  viral 
DNA  are  know  n to  cause  tumors  when  injected 
directly  into  animals,  it  was  concluded  that 
tumor  \iruses  are  noninfectious  to  animals 
when  incorporated  into  E.  coli.  If  polyoma  virus, 
which  is  the  most  infecti\  e tumor  virus  known 
for  hamsters,  cannot  cause  tumors  in  the  rDNA 
state  in  E.  coli,  it  is  unlikely  that  other  tumor 
\ iruses  w ill  do  so.  This  conclusion  has  had  wide- 
spread, but  not  unanimous,  acceptance.  It  has 
been  argued  that  there  might  be  "something 
special”  about  polyoma  that  prevents  it  from 
causing  tumors  in  this  altered  state;  other 
tumor  viruses  might  still  be  able  to  do  so.  At  one 
meeting  of  RAC,  in  fact,  it  was  suggested  that 
experiments  with  several  other  viruses  be  car- 
ried out  to  confirm  the  generality  of  the  finding. 
But  how  many  more  viruses?  What  is  enough? 


=M.  A.  Israel,  H.  VV.  Chan,  W.  P.  Rowe,  and  M.  A.  Martin,  "Molec- 
ular Cloning  of  Polyoma  V'irus  DNA  in  Escherichia  Coli:  Plasmid 
V'ector  Systems,"  Science  203:883-887,  1979. 

'Some  combinations  of  free  plasmid  and  tumor  virus  DNA  did 
cause  infections. 


202  • Impacts  of  Applied  Genetics— Micro-Organisms,  Piants,  and  Animals 


For  some,  one  carefully  planned  experiment 
using  the  most  sensitive  tests  is  sufficient  to 
allay  fears.  But  for  others,  significant  doubt 
about  safety  remains,  regardless  of  how  many 
viruses  are  examined.  The  criteria  depend  on 
an  individual’s  perception  of  risk. 

Many  experiments  carried  out  for  purposes 
other  than  risk  assessment  have  provided 
evidence  that  scenarios  of  doom  or  catastrophe 
are  highly  unlikely.  This  is  the  general  consen- 
sus of  specialists,  not  only  in  molecular  biology, 
but  in  population  genetics,  microbiology,  infec- 
tious diseases,  epidemiology,  and  public  health. 

Experiments  have  revealed  that  the  structure 
of  genes  from  higher  organisms  (plants  and 
animals)  differ  from  those  of  bacteria.  Con- 
sequently, those  genes  are  unlikely  to  be  ex- 
pressed accidentally  by  a bacterium;  the  original 
fears  of  ‘‘shotgun’’  experiments  have  become 
less  well-founded.  Hence,  data  gathered  to  date 
have  made  the  accidental  construction  of  a new 
epidemic  strain  more  unlikely. 

Conference  discussions  have  also  contributed 
to  a better  understanding  of  the  risks.  At  one 
such  conference,®  which  was  attended  by  45  ex- 
perts in  infectious  diseases  and  microbiology,  it 
was  concluded  that: 

• E.  coli  K-12  (the  weakened  form  of  E.  coli, 
used  in  experiments)  does  not  flourish  in 
the  intestinal  tract  of  man; 

• the  type  of  plasmid  permitted  by  the  Guide- 
lines has  not  been  shown  to  spread  from  E. 
coli  K-12  to  other  E.  coli  in  the  gut;  and 

• E.  coli  K-12  cannot  be  converted  to  a harm- 
ful strain  even  after  known  virulence  fac- 
tors were  transferred  to  it  using  standard 
genetic  techniques. 

A workshop  sponsored  by  NIH^  provided  a 
forum  for  scientists  to  discuss  the  risks  posed  by 
viruses  in  rDNA  experiments.  They  concluded 
that  the  risks  were  probably  less  when  a virus 
was  placed  inside  a bacterium  in  rDNA  form 

“ "Workshop  on  Studies  tor  Assessment  ol  Potential  Risks  As.soei- 
ated  With  Recomhinant  DNA  experimentation,"  I'almouth,  Mass., 
June  20-21.  1977. 

'"Workshop  to  Assess  Risks  for  Recomhinant  UNA  experiments 
Imolving  \'iral  Cenomes,"  cosponsored  hv  the  National  Institutes 
of  Health  and  the  european  Molecular  Biology  Organization, 
Ascot,  england,  Jan.  26-28,  1978. 


than  when  it  existed  freely.*  Experts  in  infec- 
tious disease  have  stressed  repeatedly  that  the 
ability  of  a micro-organism  to  cause  disease 
depends  on  a host  of  factors,  all  working  togeth- 
er. Inserting  a piece  of  DNA  into  a bacterium  is 
unlikely  to  suddenly  transform  the  oi'ganism 
into  a virulent  epidemic  strain. 

Careful  calculations  can  also  allay  fears  about 
the  damage  a genetically  engineered  micro-or- 
ganism might  cause.  Doomsday  scenarios  of 
escaped  E.  coli  that  carry  insulin  or  other 
hormone-producing  genes  were  recently  exam- 
ined in  another  workshop.®  I’rior  to  this  work- 
shop, newspaper  accounts  raised  the  possibility 
that  an  E.  coli  carrying  the  gene  for  human  in- 
sulin production  might  colonize  humans  and 
thus  upset  the  hormonal  balance  of  the  body. 

The  participants  calculated  how  much  insulin 
could  be  produced.  First,  it  was  assumeil  that  a 
series  of  highly  unlikely  events  would  occur— 
accidental  release,  ingestion  by  humans,  stable 
colonization  of  the  intestine  by  E.  coli  K-12.  E. 
coli  constitutes  approximately  1 percent  of  tlu? 
intestinal  bacterial  population,  and  it  was 
assumed  that  all  the  normal  E.  coli  would  h(* 
replaced  by  the  insulin-producing  E.  coli.  Insulin 
is  made  in  the  foi-m  of  a precursor  moh'cule, 
proinsulin.  It  was  assumed  that  50  p(>rcent  of  all 
bacterial  protein  [jroduction  would  h(>  dexoted 
to  this  single  pi'Otein,  anotluM-  highly  unlikely 
situation.  If  so,  30  micrograms  (;ig)— or  0 0 
units— would  then  he  made  in  the  inlestiiuv 
Although  proteins  are  \’(M’v  |)oorly  ai)sorhed 
from  the  intestinal  ca\  ity,  it  w as  assumed  foi' 
the  sake  of  argument  that  100  percent  of  the 
proinsulin  would  h(f  absorbed  into  the  circula- 
tion. Thus,  0.0  units  of  insulin  would  he  added 
to  the  noi’mal  dail\’  human  production  of  25  to 
30  units— an  imperceptible  difference. 

Calculations  like  these  ha\c  been  cai  i ied  «)Ut 
for  several  other'  hoi  inones.  I',\(*n  with  the  most 
implausible  seri(\s  of  (wents,  leading  to  the 
gi'eatest  oppoi’tunity  for  hormone  pi'oduction. 

'On  Iho  ()lh(‘r  hand.  Il  h.is  hrnn  .irgucil  lhal  ihiv  ha\  |iiii\idi-a 
vim.so.s  with  a new  mule  lor  (li.sM'ininalion  Nf\ ri  ihrlcss  thi-n-  i-. 
no  cvidiMU'C  that  v irn.scs  can  icadih  cm  .ipr  li  om  the  li.n  in  i.i  .mil 
.snh.s(HHi('nllv  cau,s(‘  inicclion 

"'"National  Inslilulc  ol  Mlcrg^v  .ind  Inicc  lions  I Iim  .im-s  v\  m k .liup 
on  Recomhinant  l).\  \ Risk  \sscssincnl  I’.is.idcn.i  ( .ilil  \|n 
11-12.  1981) 


Ch.  10 — The  Question  of  Risk  • 203 


the  c'oiK'lusion  is  that  noi'mal  hormone  le\els 
would  change  by  less  than  10  percent.  Similar 
coiulitions  toi’  interferon  production  could 
release  a[)pro.\imately  70/ig  or  the  ma.ximum 
dail\  dose  currently  used  in  cancer  therapy, 
l.ong-term  effects  of  such  e.xposure  ai'e  current- 
ly unknow  ti:  therefore,  experiments  using  high- 
producing  strains  (10®  molecules  per  cell  or 
more)  aiv  likely  to  he  monitored  if  such  strains 
e\  er  hecome  a\  ailahle. 

The  .\IH  program  of  risk  assessment,  which 
was  formalK  started  in  1979,  continues  to  iden- 
tity possible  consetiuences  of  rDN'.A  research. 
L'nder  the  aegis  of  the  National  Institute  of 
AllergN’  and  Infectious  Diseases,  the  progi'am 
supports  research  studies  designed  to  elucidate 
the  likelihood  of  harm.’  In  addition,  it  collates 
general  data  from  other  experiments  that  might 
he  rele\ant  to  risk  assessment.  Other  risk  as- 
sessments are  being  conducted  by  European 
organizations”  and  by  the  L'.S.  Environmental 
Protection  .-\genc\'  to  assess  the  consec|uences  of 
releasing  micro-organisms  into  the  en\iron- 
ment. 

Thus  far.  there  is  no  compelling  ex  idence  that 
E.  coli  K-12  bacteria  carrying  rDN.A  will  be  more 
' hazardous  than  any  of  the  micro-organisms 
I which  serxed  as  the  source  of  D\,A.  Nexer- 
! theless,  all  the  experiments  hax  e dealt  with  one 
I genus  of  bacterium.  Unless  the  conclusions 
about  £.  coli  can  be  extended  to  other  organisms 
likely  to  be  used  in  experiments  (such  as  Bacillus 
subtilis  and  yeast),  other  assessments  may  be  ap- 
propriate. 


’E.\tramural  efforts  were  first  conceived  in  the  summer  of  1975 
to  develop  and  test  safer  host-vector  systems  based  on  £.  coli,  the 
interagency  agreement  entered  into  with  the  ,\a\al  Biosciences 
Laboratory  tested  £.  coli  systems  in  a series  of  simulated 
accidental  spills  in  the  laboratory.  .Xt  the  Uni\ersity  of  .Xlichigan 
the  survival  of  these  systems  was  tested  in  mice  and  in  cultural 
conditions  simulating  the  mouse  gastrointestinal  tract.  Tufts 
Lniversity  tested  these  systems  in  both  mice  and  human 
volunteers.  Finally,  the  surv  ival  of  host-\ector  systems  in  sewage 
treatment  plants  was  tested  at  the  Unh  ersity  of  Te.xas.  The  peak 
year  for  costs  of  supporting  research  contracts  was  1978;  over  a 
half-million  dollars  were  required.  Currently,  the  cost  of 
maintaining  the  high  containment  facility  at  Frederick,  Md.,  is 
between  S200.000  and  S250.000  annually. 

••First  Report  to  the  Committee  on  Genetic  E.\perimentation  . a 
scientific  committee  of  the  International  Council  of  Scientific 
Unions,  from  the  Working  Group  on  Risk  .Assessment,  July  1978. 


Perception  of  risk. 

Tbe  probability  of  damage  can  be  estimated 
for  xarious  exents.  Tbe  entire  insurance  in- 
dustry is  based  on  the  fact  that  unfavorable 
exents  occur  on  a regular  basis.  The  number  of 
people  dying  annually  from  cancer,  or  automo- 
bile accidents,  or  homicides  can  he  predicted 
fairly  accurately.  These  estimates  depend  on 
the  ax  ailahility  of  data  and  the  assumptions  that 
the  major  determinants  do  not  change  from 
year  to  year. 

But  ex  en  if  the  probability  of  damage  is  fairly 
well  knoxxn,  a gap  often  exists  between  this 
"real”  probability  of  occurrence  and  the  "per- 
ceixed”  probability.  Txxo  factors  that  tend  to  af- 
fect perceptions  are  the  magnitude  of  the  possi- 
ble damage  and  the  lack  of  individual  control 
ox  er  exposure  to  the  risk.  Both  of  these  are  sig- 
nificant factors  in  the  fears  associated  xvith 
rDN.A  and  the  manipulation  of  genes.  Because 
intuitixe  exaluations  can  contradict  analytical 
exaluations,  the  question  of  risk  cannot  be  re- 
solx  ed  strictly  on  an  analytical  basis.  Its  resolu- 
tion xx'ill  have  to  come  through  the  political 
process. 

BURDEN  OF  PROOF 

The  possibility  of  inadvertently  creating  a 
dangerous  organism  does  exist,  but  its  prob- 
ability is  lower  than  was  originally  thought. 
Nevertheless,  an  important  principle  emerges 
from  the  debate.  Society  must  decide  whether 
the  burden  of  proof  rests  xvitb  those  who  de- 
mand evidence  of  safety  or  with  those  who  de- 
mand evidence  of  hazard.  The  former  would 
halt  experiments  until  they  are  proved  safe.  The 
latter  xvould  continue  experiments  until  it  is 
shown  that  they  might  cause  harm. 

A significant  theoretical  difference  exists  be- 
txveen  the  tw  o approaches.  Evidence  can  almost 
alxx^ays  be  provided  to  show  that  something 
causes  harm— e.g.,  it  can  be  demonstrated  that  a 
poliovirus  causes  paralysis,  that  a Pneumococcus 
causes  pneumonia,  that  a rhinovirus  causes  the 
common  cold.  However,  it  cannot  be  demon- 
strated that  a poliovirus  can  never  cause  the 
common  cold.  It  cannot  be  demonstrated  that 
rDNA  molecules  will  never  be  harmful.  It  can 


204  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


only  be  demonstrated  that  harmful  events  are  level  of  uncertainty  it  is  willing  to  accept. 
unlikely.  Hence,  society  must  determine  what 


Other  concerns  

Concerns  raised  by  industrial 
applications 

Originally  concerns  involved  hazards  that 
might  arise  in  the  laboratory.  Now  that  there 
are  industrial  applications  of  genetic  engineer- 
ing, the  concerns  include: 

• risks  associated  with  the  laboratory  con- 
struction of  new  strains  of  organisms, 

• risks  associated  with  industrial  production 
or  consumer  use  of  the  new  strains,  and 

• risks  associated  with  the  products  obtained 
from  the  new  strains. 

Many  similar  considerations  apply  to  the  as- 
sessment of  the  first  two  kinds  of  risks.  Unless 
the  organisms  used  in  an  industrial  production 
scheme  are  thoroughly  characterized,  conjec- 
tured fears  about  their  ability  to  cause  disease 
will  continue.  Even  with  a recombinant  orga- 
nism that  has  a well-defined  sequence  of  DNA,  a 
break  in  containment  would  leave  its  behavior 
in  the  environment  questionable.  Experience 
with  substances  such  as  asbestos  gives  rise  to 
fears  that  exposure  to  the  new  biological  sys- 
tems might  also  cause  unforseen  pathological 
conditions  at  some  future  time. 

Hazards  associated  with  products  raise  dif- 
ferent questions.  The  growing  consensus  in 
Federal  regulatory  agencies  appears  to  be  that 
these  products  should  be  assessed  like  all 
others— e.g.,  human  growth  hormone  (hGH) 
produced  by  genetically  engineered  bacteria 
should  be  tested  for  purity,  chemical  identity, 
and  biological  activity  just  like  hGH  from  human 
pituitary  glands.  The  possibility  of  product 
variation  due  to  mutation  of  the  bacteria, 
however,  suggests  that  batch  testing  and  certifi- 
cation might  be  warranted  as  well.  (For  further 
discussion  see  ch.  11.) 


Concerns  raised  by  the  implications  of 
the  rDNA  controversy  for  general 
microbiology 

Questions  about  the  |)Otential  hai'in  from 
genetically  engineered  micro-organisms  have 
led  to  questions  about  the  efforts  curixMitly 
employed  to  protect  the  public  fi'om  work  being 
done  with  micro-organisms  known  to  he  hazard- 
ous. These  viruses,  bacteria,  and  fungi  are 
handled  daily  in  laboratory  expei’iments,  in  the 
routine  isolation  of  infectious  agents  from  |)a- 
tients,  and  in  the  production  of  \ accines  in  the 
pharmaceutical  industry. 

Questions  have  been  raised  about  the  efficacy 
of  regulations  established  for  these  xai'ious 
potentially  hazardous  agents.  A full-s('ale  as.sess- 
ment  is  not  within  the  scope  ot  this  study,  hut  it 
is  clear  that  the  (|uestions  are*  piM  tiniMit.  I wo 
conclusions  have  been  reaclKul. 

First,  there  is  a growing  h(‘li('f  that  the  mere 
existence  of  a classification  scheme  for  ha/.aid- 
ous  agents  by  the  Clenter  for  Disease  (l)nlrol 
(GDC)  is  not  enough  to  ensure  their  .safe  han- 
dling. The  Subcommittee  on  .\rho\  irus  Labora- 
tory Safety  was  formed  I'ecently  because  of  con- 
cerns expressed  in  academic  ('ircles.  Ke|)ia‘senl- 
atives  from  unixersities,  the  I’uhlic  Health  Serv- 
ice, the  U.  S.  DepartiiHMit  of  .Agriculture,  and 
the  military,  who  constitutc'd  the  suhcommit- 
tee,  are  prej)aring  a report  based  on  an  interna- 
tional survey  of  laboratory  practices  and  inlec 
tions.  They  found  wide'  vaiiation  in  the  wavs 
different  agents  vv(M'(>  liandled  Most  ol  their 
recommendations  are  idcMitical  with  those  ,i|)- 
plicahle  to  rDNA— that  appropriate  cont.iinment 
levels  he  used  with  diffeicnt  viiuses  that  the 
health  of  workers  lu'  monitored,  and  th.it  .in  In 
stitutional  Biosafety  (l)mmittee  he  appointed  to 
serve  each  institution. 


Ch.  10 — The  Question  of  Risk  • 205 


Second,  little  is  known  ahoiit  the  health 
record  ot  v\orkers  in\  oKed  iti  the  fermentation 
and  vaccine  industries.  Foi'  most  industrial 
operations  the  e\  idence  of  harm  is  almost  en- 
tirely anecdotal.  .Most  industrial  fermentations 
are  regarded  as  hai  mless:  representativ  es  of  in- 
dustry characterize  it  as  a "non-pmhlem”  that 
has  never  merited  monitoring.  Conifirehensive 
information  on  the  potential  harmful  effects 
associated  w ith  research  using  rn\.\-carrying 
micro-organisms  w ill  not  he  available  because 
the  (iuidelines  consider  it  the  responsibility  of 
each  institution  or  companv  to  "determine,  in 
connection  with  each  project,  the  necessitv  for 
medical  sui'v  eillance  of  recomhinant-l),\,\  re- 
search personnel."  Hence  some  institutions 
might  decide  to  keep  records  of  some  or  all  ac- 
tiv  ities;  others  might  not. 

To  he  sin  e,  some  companies  have  e.xceeded 
the  minimal  medical  standards  set  by  \'1H  for 
fermentation  using  rDN.A-carrv  ing  micro-orga- 
nisms—eg.,  Kli  l.illy  &.  Co.  requires  that  all 
illnesses  he  reported  to  supervisors  and  that  any 
employees  who  are  ill  for  more  than  5 days 
must  report  to  a phvsician  before  being  allowed 
to  return  to  work.  .Any  employee  taking  antibi- 
otics (vv  hich  might  make  it  easier  for  bacteria  to 
colonize)  is  restricted  from  areas  where  rDN.A 
research  is  being  done  until  5 days  after  the  dis- 
continuance of  the  antibiotic.  .At  .Abbott  Labora- 
tories. a physician  checks  into  the  illness  of  any 
recombinant  worker  who  is  off  more  than  1 
dav— a precaution  taken  onlv  after  5 days  off 
for  workers  in  other  areas.  Lilly  maintains  a 
computer  listing  of  all  workers  involved  in 
rD.VA  activities.  Lilly,  the  Upjohn  Co.,  and 
Merck,  Sharp  and  Dohme  have  been  in  the 
process  of  computerizing  the  health  records  of 
all  their  employees  over  the  past  several  years. 

Work  with  rD\A  has  focused  attention  on 
biohazards  and  medical  surveillance— an  aware- 
ness that  had  arisen  in  the  past  but  had  not  been 
sustained.*  Consequently,  several  documents 
on  the  subject  either  have  been  or  will  be  pub- 
lished: 


■.As  of  Sepiember  1980,  the  .National  Institutes  of  Occupational 
Safety  and  Health  and  the  environmental  Protection  Agency  were 
planning  to  fund  a.ssessnients  of  the  adequacy  of  current  medical 
surv  eillance  technologv-. 


• CDC  is  preparing  a complete  revision  of  its 
laboratory  safety  manual,  wdiich  is  widely 
used  as  a starting  point  by  other  labora- 
tories. 

• The  Classification  of  Etiologic  Agents  on  the 
Basis  of  Hazard,  which  was  last  revised  in 
1974,  has  been  expanded  by  CDC  in  collab- 
oration with  NIH  into  a Proposed  Biosafety 
Guidelines  for  Microbiological  and  Biomedi- 
cal Laboratories.  These  guidelines  serve  the 
purpose  tultilled  by  the  Dangerous  Patho- 
gens Advisory  Group  (DRAG)  in  the  United 
Kingdom,  although  they  lack  any  regula- 
tory strength. 

• A comprehensive  program  in  safety, 
health,  and  environmental  protection  was 
developed  in  1979  by  and  for  NIH.  It  is  ad- 
ministered by  the  Division  of  Safety,  which 
includes  programs  in  radiation  safety,  oc- 
cupational safety  and  health,  environmen- 
tal protection,  and  occupational  medicine. 

• The  Office  of  Biohazard  Safety,  National 
Cancer  Institute  has  just  completed  a 3- 
year  study  of  the  medical  surveillance  pro- 
grams of  its  contractors;  a report  is  being 
drafted. 

Although  the  academic,  governmental,  and 
industrial  communities  have  shown  growing  in- 
terest in  biosafety,*  no  Federal  agency  regulates 
the  possession  or  use  of  micro-organisms  except 
for  those  highly  pathogenic  to  animals  and  for 
interstate  transport.**  Whether  such  regula- 
tions are  necessary  is  an  issue  that  extends  be- 
yond the  scope  of  this  study.  Nevertheless, 
other  countries— for  instance  the  United  King- 
dom, with  its  DPAG— have  acted  on  the  issue. 
This  organization  functions  specifically  to  guard 
against  hazardous  micro-organisms,  by  moni- 
toring and  licensing  university  and  industrial 
laboratories  and  meting  out  penalties  when 
necessary. 


•Curiously,  there  is  no  formal  society  or  journal,  but  there  has 
been  an  annual  Biological  Safety  Cionference  since  1955,  con- 
ducted on  a round-robin  basis  primarily  by  close  associates  of  the 
late  Arnold  VV'edum,  M.D.— former  Director  of  Industrial  Health 
and  Safetvat  the  I'.S,  Army  Biological  Research  Laboratories,  Fort 
Detrick,  Md..  who  is  regarded  as  the  "Father  of  Microbiological 
Safety." 

*”ln  some  States  and  cities,  licensing  is  required  for  all  facilities 
handling  pathogenic  micro-organisms. 


206  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Concerns  raised  by  the  implications  of 
the  rDNA  controversy  for  other 
genetic  manipulation 

Altering  the  hereditary  characteristics  of  an 
organism  by  using  rDNA  is  just  one  of  the 
several  methods  of  genetic  engineering.  The 
definition  of  rDNA  refers  specifically  to  the 
combination  of  the  DNA  from  two  organisms 


outside  the  cell.  If  the  DNA  is  combined  within 
living  cells,  the  Guidelines  do  not  pertain.  Figure 
35  shows  several  methods  that  achieve  the  same 
goal— transfering  genetic  material  from  one  cell 
to  another,  bypassing  the  normal  se.xual 
mechanisms  of  mating.  It  is  particulaiiy  signifi- 
cant that  DNA  from  different  sftecies  can  he 
combined  by  all  these  mechanisms,  only  one  of 
which  is  rDNA.  Different  species  of  bacteria, 


Figure  37.— Alternative  Methods  for  Transferring  DNA  From  One  Cell  to  Another 


A.  The  two  cells  are  fused  in  toto 

B.  A microcell  with  a fragmented  nucleus  carries  the  DNA 

C.  Free  DNA  can  enter  the  recipient  cell  in  a number  of  ways;  by  direct  microinjection,  through 
calcium-mediated  transformation,  or  by  being  coated  with  a phospholipid  membrane  in 
order  to  fuse  with  the  recipient  cell 

D.  The  free  DNA  can  be  joined  to  a plasmid  and  transferred  as  recombinant  DNA 


SOURCE:  Office  of  Technology  Assessment. 


Ch.10 — The  Question  of  Risk  • 207 


fungi,  and  higher  organisms  can  all  he  fused  or 
manipulated.  * 

Opponents  of  rDX.A  ha\e  stated  that  combin- 
ing genes  from  different  species  may  disturb  an 
e.xtremely  intricate  ecological  interaction  that  is 
onl\'  dimly  understood.  Hence,  such  e.xperi- 
ments,  it  is  argued,  are  unpredictable  and  there- 
fore hazardous.  If  so,  all  the  other  methods 
represented  in  figure  35  should  he  included  in 
the  (iuidelines.  \ et  they  are  not. 

rhe  most  acceptable  e.xplanation  for  this  in- 
consistency is  that  rl),\.\  is  currenth’  the  most 

*Kor  example,  anlihiotie  reNi.siant  pla.smicls  ha\e  been  tran.>;- 

I'eiTtHl  from  Staphylocotxus  aurvus  to  Radllus  suhtilis  acros.s 
sp«'eies  barriers  by  transtormation,  not  by  rON  A.  Foreign  genes 
for  the  enzyme  amyla.s**  hax  e al.so  been  inlixHlueed  into  fl.  sublilis. 


efficient  and  successful  method  of  combining 
genes  from  \ ery  diverse  organisms.  It  is  reason- 
able to  ask,  however,  what  would  happen  if  any 
of  the  other  methods  become  equally  success- 
ful. Will  a profusion  of  guidelines  appear?  Will 
one  committee  oversee  all  genetic  experiments 

Ethical  and  moral  concerns 

The  perceixed  risk  associated  with  genetic 
engineering  includes  ethical  and  moral  hazards 
as  well  as  physical  ones.  It  is  important  to 
recognize  that  these  are  part  of  the  general 
topic  of  risk.  To  some,  there  is  just  as  much  risk 
to  social  values  and  structure  as  to  human 
health  and  the  environment.  (For  further  dis- 
cussion see  ch.  13.) 


Conclusion  

Thus  far,  no  demonstrable  harm  associated 
with  genetic  engineering,  and  particularly 
rDX.A,  has  been  found.  But  although  demonstra- 
ble harm  is  based  on  e\  idence  that  damage  has 
occurred  at  one  time  or  another,  it  does  not 
mean  that  damage  cannot  occur. 

Conjectural  hazards  based  on  analogies  and 
scenarios  ha\e  been  addressed  and  most  ha\  e 
proxed  less  xxorrisome  than  prexiously  as- 
sumed. Xexertheless,  there  is  agreement  that 
certain  experiments,  such  as  the  transfer  of 
genes  for  knoxvn  toxins  or  x enoms  into  bacteria, 
should  still  be  prohibited  because  of  the  real 
likelihood  of  danger.  Still  other  experiments 
cannot  clearly  be  shoxvn  to  be  hazardous  or 
readily  dismissed  as  harmless.  Hence,  a political 
decision  is  likely  to  be  required  to  establish 
xvhat  constitutes  acceptable  proof  and  xvho 
must  prox  ide  it. 

Gix  en  that  potential  harm  can  be  identified  in 
some  cases,  its  probable  occurrence  and  magni- 
tude quantified,  and  perceived  risk  taken  into 
account,  a decision  to  proceed  is  usually  based 
on  society’s  xvillingness  to  take  the  risk.  This 
triad  of  the  physical  {actual  risk),  psychological 
{perception  of  risk),  and  political  {willingness  to 
take  risk)  plays  a role  in  all  decisions  relating  to 
genetic  engineering. 


The  potential  benefits  must  always  be  con- 
sidered along  with  the  risks.  Decisions  made  by 
RAC  haxe  reflected  this  view— e.g.,  when  it 
approx'ed  the  cloning  of  the  genetic  material  of 
the  foot-and-mouth  disease  virus.  The  perceived 
benefits  to  millions  of  animals  outweighed  the 
potential  hazard. 

Recombinant  DNA  techniques  represent  just 
one  of  several  methods  to  join  fragments  of 
DNA  from  different  organisms.  The  current 
Guidelines  do  no  extend  to  these  other  tech- 
niques, although  they  share  some  of  the  same 
uncertainties.  Ignoring  the  consequences  of  the 
other  technologies  might  be  viewed  as  an  incon- 
sistency in  policy- 

while  the  initial  concerns  about  the  possibili- 
ty of  hazards  at  the  laboratory  level  appear  to 
have  been  overstated,  other  types  of  potential 
hazards  at  different  stages  of  the  technology 
have  been  identified.  Emphasis  has  shifted 
somewhat  from  conjectured  hazards  that  might 
arise  from  research  and  development  to  those 
that  might  be  associated  with  production  tech- 
nologies. As  a consequence,  there  is  a clearer 
mandate  for  existing  Federal  regulatory  agen- 
cies to  play  a role  in  ensuring  safety  in  industrial 
settings. 


chapter  1 1 

Regulation  of 
Genetic  Engineering 


chapter  11 


Page 


Introduction 211 

Framework  for  the  Analysis 211 

Current  Regulation:  the  NIH  Guidelines 212 

Substantive  Requirements 212 

Administration 212 

Provisions  for  Voluntary  Compliance 215 

Evaluation  of  the  Guidelines 216 

The  Problem  of  Risk 216 

The  Decisionmaking  Process 221 

Conclusion 223 

Other  Means  of  Regulation 224 

Federal  Statutes 224 

Tort  Law  and  Workmen’s  Compensation 227 


State  and  Local  Law 229 

Conclusion 230 

Issue  and  Options 230 


Tables 

Table  No.  Page 

35.  Containment  Recommended  l)v  National 

Institutes  of  Health 213 

36.  Statutes  That  Will  Be  Most  Applicable  to 

Commercial  Genetic  Engineering 224 


Chapter  11 

Regulation  of  Genetic  Engineering 


Introduction  

Although  no  e\  idence  exists  that  any  hai'mful 
organism  has  been  created  hv  molecular  genetic 
techniques,  most  e\})erts  helie\e  that  some 
risk*  is  associated  with  genetic  engineering. 
One  kind  is  relatively  certain  and  ciuantitiable— 
that  of  working  with  known  toxins  or  patho- 
gens. .Another  is  uncertain  and  hypothetical— 
that  of  the  possible  creation  of  a [)athogenic  or 
otherwise  undesirable  organism  by  reshuffling 
genes  thought  to  he  harmless.  These  may  he 
thought  of  as  physical  risks  because  they  con- 
cern human  health  or  the  en\  ironment. 

(x)ncern  has  also  arisen  about  the  possible 
long-range  impacts  of  the  techni(|ues— that  they 
may  eventually  he  used  on  humans  in  some 
morally  unacceptable  manner  or  may  change 
fundamental  \ iews  of  w hat  it  means  to  be  hu- 
man. These  possibilities  may  he  thought  of  as 
cultural  risks,  since  they  threaten  fundamental 
beliefs  and  v alue  systems.' 

The  issue  of  whether  or  not  to  regulate 
molecular  genetic  techniques— and  if  so,  to 
what  extent— defies  a simple  solution.  Percep- 
tions of  the  nature,  magnitude,  and  acceptabili- 
ty of  the  risks  differ  drastically.  Approximately 
6 years  ago,  vv  hen  the  scientific  community  it- 
self accepted  a moratorium  on  certain  classes  of 
recombinant  DX.A  (rD\,A)  research,  some  sci- 
entists considered  the  concern  unnecessary.  To- 
day, even  though  the  physical  risks  of  rDNA  re- 
search are  generally  considered  to  be  less  than 
originally  feared— and  the  realization  of  its 
benefits  much  closer— some  people  would  still 
prohibit  it. 

The  Federal  Government's  approach  to  this 
issue  has  been  the  promulgation  of  the  Guide- 
lines for  Research  Involving  Recombinant  DNA 
Molecules  (Guidelines),  by  the  National  Insti- 
tutes of  Health  (NIH).  (See  app.  III-C  for  infor- 
mation about  what  other  countries  have  done 

" As  used  in  this  chapter,  risk  means  the  possibility  of  harm.  The 
probabilitv  of  that  harm  occurring  may  be  e.xtremelv  low  and/or 
highly  uncertain. 

'H.  Tristam.  Engelhardt.  Jr.,  “Taking  Risks:  Some  Background 
Issues  in  the  Debate  Concerning  Recombinant  D,\'.A  Research, 
Southern  California  Law  Review  o\:6,pp.  1141-1151.  1978. 


with  respect  to  guidelines  for  rDNA.)  Three 
other  available  modes  of  oversight  or  regulation 
are  current  Federal  statutes,  toi't  law,  and  State 
and  local  law. 

Frameworh for  the  analysis 

In  deciding  how  to  address  the  risks  posed  by 
genetic  engineering,  some  of  the  important 
questions  that  need  to  be  examined  are; 

• How  broadly  the  scope  of  the  issue  (or 

problem)  should  be  defined. 

—Who  identifies  the  risks  and  their  mag- 
nitude? 

—Who  proposes  the  means  for  addressing 
the  problem? 

• The  nature  of  the  procedural,  decisionmak- 
ing mechanism. 

—Who  decides? 

—Who  will  benefit  from  the  proposed  ac- 
tion and  who  will  bear  the  risk? 

—Will  the  risk  be  borne  voluntarily  or  in- 
voluntarily? 

—Who  has  the  burden  of  proof? 

— SboLild  a risk/benefit  analysis,  or  some 
other  approach,  be  used? 

• The  available  solutions  and  their  adequacy. 

—Should  there  be  full  regulation,  no  reg- 
ulation, or  something  in-between? 

—What  actions  and  actors  should  be  cov- 
ered? 

—What  is  the  appropriate  means  for  en- 
forcing a regulatory  decision? 

—Which  agency  or  other  group  should  do 
the  regulating? 

Underlying  these  questions  is  the  proposition, 
widely  accepted  by  commentators  on  science 
policy,  that  scientists  are  qualified  to  assess 
physical  risk,  since  that  inyolyes  measuring  and 
evaluating  technical  data.  Howeyer,  a judgment 
of  safety  (the  acceptability  of  that  risk)  can  only 
be  made  by  society  through  the  political  proc- 
ess, since  it  involyes  weighing  and  choosing 
among  yalues.^  3 4 5 6 Scientists  are  not  nec- 

^VVilliam  VV.  Lowrance,  Of  Acceptable  Risk:  Science  and  the  De- 
termination of  Safety  (Los  Altos,  C;alit.:  William  Kaufmann,  Inc., 
1976). 


211 


212  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


essarily  considered  to  be  more  qualified  to  make 
decisions  concerning  social  values  than  other 
well-informed  persons;  they  may  in  fact  be  less 
qualified  when  the  decision  involves  possible 
restrictions  on  scientific  research  because  of  the 

IconliniK'd  from  p.  211) 

'Alvin  \V.  VVninIxM'f',  "Scitxirc  and  Irans-ScifMici!,''  MinorvH, 
10:2,  April  li)72. 

^Allan  Mazur,  Oisputes  Between  Kxpei  ts,”  Minerva  11:2,  April 
1973. 

"Arthur  Kanti-owitz,  "The  Science  Court  Experiment,"  Juri- 
metrics  Journal,  vol.  17,  1977,  p.  332. 

“David  E.  Bazelon,  "Risk  and  Responsibility,"  Science,  vol.  205, 
July  20,  1979,  pp.  277-280. 


high  value  they  place  on  unrestricted  research 
and  because  of  possible  conflicts  of  interest. 
Moreover,  according  to  this  view,  if  society  is  to 
bear  a risk,  it  should  judge  the  acceptability  of 
that  risk  and  give  its  informed  consent  to  it.^* 


^Engelhard!,  op.cil.;  Eowi'ance,  op.cit.:  and  Bazelon,  op.  cit. 

•In  practice,  it  may  olten  be  diUicult  to  keep  the  two  kiiuls  of 
decisions  separate,  since  the  values  of  indix  idual  scientists  may  in- 
fluence their  interpi-etation  of  technical  data,  and  since  policy- 
makers may  not  have  the  technical  competence  to  understand  the 
I’isks  sufficiently." 

"Weinberg,  op.  cit.:  and  Bazelon,  op.  cit. 


Current  regulation:  the  NIH  Guidelines 


The  Guidelines  have  been  developing  in 
stages  over  a period  of  approximately  6 years  as 
scientists  and  policymakers  have  grappled  with 
the  risks  posed  by  rDNA  techniques.  (This  his- 
tory, discussed  in  app.  III-A,  is  crucial  to  under- 
standing current  regulatory  issues,  and  it  serves 
as  a basis  for  evaluating  the  Guidelines.)  They 
represent  the  only  Federal  oversight  mecha- 
nism that  specifically  addresses  genetic  engi- 
neering. 

Substantive  requirements 

The  Guidelines  apply  to  all  research  involving 
rDNA  molecules  in  the  United  States  or  its  ter- 
ritories conducted  at  or  sponsored  by  any  in- 
stitution receiving  any  support  for  rDNA  re- 
search from  NIH.  Six  types  of  experiments  are 
specifically  prohibitedi)  1)  the  formation  of 
rDNA  derived  from  certain  pathogenic  orga- 
nisms; 2)  the  formation  of  rDNA  containing 
genes  that  make  vertebrate  toxins;  3)  the  use  of 
the  rDNA  techniques  to  create  certain  plant 
pathogens;  4)  transference  of  drug  resistance 
traits  to  micro-organisms  that  cause  disease  in 
humans,  animals,  or  plants;  5)  the  deliberate 
release  of  any  organism  containing  rDNA  into 
the  environment;  and  6)  experiments  using 
more  than  10  liters  (1)  of  culture  unless  the 
rDNA  is  “rigorously  characterized  and  the 
absence  of  harmful  sequences  established.’^  A 
procedure  is  specified  for  obtaining  exceptions 


from  these  prohibitions.  Five  types  of  experi- 
ments are  completely  exempt. 

Those  experiments  that  are  neithffr  prohib- 
ited nor  exempt  must  he  carried  on  in  ac- 
cordance with  physical  and  biological  contain- 
ment levels  that  relate  to  the  degree  of  potential 
hazard.  (See  table  33.)  Physical  containment  re- 
quires methods  and  eciuipment  that  Ufssen  the 
chances  that  a recombinant  organism  might  es- 
cape. Four  levels,  designated  FI  for  thi^  k’ast 
restrictive  through  F4  for  the  most,  are  defined. 
Biological  containment  recjuires  working  witli 
weakened  organisms  that  are  unlikedy  to  sur- 
vive any  escape  from  the  laboratory,  three 
levels  are  specified.  Glasses  of  pcMinitted  e.x- 
periments  are  assigned  l)oth  |)hysical  and  bio- 
logical containment  levels.  Most  experiments 
using  Escherichia  coli  K-12,  th(^  standard  lal)ora- 
tory  bacterium  used  in  appro.ximately  «()  per- 
cent of  all  exjjeriments  co\(!red  by  the  (Guide- 
lines, may  be  perfornuHl  at  tlu*  low(‘st  contain- 
ment levels. 

AIIMINISTHA'I  I(>\ 

The  Guidelines  pi'o\  ide  an  administrativ r 
framework  foi'  implementation  that  specifies 
the  roles  and  I’esponsihilities  of  the  scientists, 
their  institutions,  and  the  I'ederal  Government. 
The  parties  who  are  crucial  to  the  effective 
operation  of  the  system  are:  1)  the  Director  ol 
NIH,  2)  the  NIH  Hecomhinant  D.VA  Advisory 
Committee  (RA(3,  3)  the  Mil  Office  ol  Itecomhi 


Ch.  11 — Regulation  of  Genetic  Engineering  • 213 


Table  35.  — Containment  Recommended  by 
National  Institutes  of  Health 


Biological — Any  combination  of  vector  and  host  must  be 
chosen  to  minimize  both  the  survival  of  the  system 
outside  of  the  laboratory  and  the  transmission  of  the 
vector  to  nonlaboratory  hosts.  There  are  three  levels 
of  biological  containment; 

HV1—  Requires  the  use  of  Escherichia  coli  K12  or 

other  weakened  strains  of  micro-organisms  that 
are  less  able  to  live  outside  the  laboratory. 

HV2—  Requires  the  use  of  specially  engineered  strains 
that  are  especially  sensitive  to  ultraviolet  light, 
detergents,  and  the  absence  of  certain 
uncommon  chemical  compounds. 

HV3—  No  organism  has  yet  been  developed  that  can 
qualify  as  HV3. 

Physical — Special  laboratories  (P1-P4) 

PI—  Good  laboratory  procedures,  trained  personnel, 
wastes  decontaminated. 

P2—  Biohazards  sign,  no  public  access,  autoclave  in 
building,  hand  washing  facility. 

P3—  Negative  pressure,  filters  in  vacuum  line,  class  II 
safety  cabinets. 

P4—  Monolithic  construction,  air  locks,  all  air 
decontaminated,  autoclave  in  room,  all 
experiments  in  class  III  safety  cabinets  (glove 
box),  shower  room. 


SOURCE;  Office  of  Technology  Assessment 


nant  DX.A  ,Acti\ities  (ORDA),  4)  the  Federal  In- 
teragency Ad\  isorv  Committee  on  Recombinant 
DN'.A  Research  (Interagency  Committee),  5)  the 
Institution  where  the  research  is  conducted,  6) 
the  Institutional  Biosafety  Committee  (IBC),  7) 
the  Principal  In\estigator  (PI),  and  8)  the  Bio- 
logical Safety  Officer. 

The  Director  of  t\IH  carries  the  primary  bur- 
den for  the  Federal  Go\  ernment’s  oversight  of 
rDNA  activities,  since  he  is  responsible  for  im- 
plementing and  interpreting  the  Guidelines,  es- 
tablishing and  maintaining  R.AC  (a  technical  ad- 
\ isorv  committee)  and  ORDA  (whose  functions 
are  purely  administrath  e),  and  maintaining  the 
Interagency  Committee  (which  coordinates  all 
Federal  acti\ities  relating  to  rDNA).  Under  this 
arrangement,  all  decisions  and  actions  are  taken 
by  the  Director  or  his  staff.  For  major  actions, 
the  Director  must  seek  the  advice  of  RAC,  and 
he  must  provide  the  public  and  other  Federal 
agencies  with  at  least  30  days  to  comment  on 


proposed  actions.  Such  actions  include:  1) 

assigning  and  changing  containment  levels  for 
e.\j)eriments,  2)  certifying  new  host-vector  sys- 
tems, 3)  maintaining  a list  of  rDNA  molecules  ex- 
empt from  the  Guidelines,  4)  permitting  excep- 
tions to  prohibited  experiments,  and  5)  adopting 
changes  in  the  Guidelines. 

For  other  specified  actions,  the  Director  need 
onh'  inform  R.AC,  the  IBC's,  and  the  public  of  his 
decision.  The  most  important  of  tliese  are:  1) 
making  minor  interpretive  decisions  on  contain- 
ment for  certain  experiments;  2)  authorizing, 
under  procedures  specified  by  RAC,  large-scale 
work  (in\'ol\  ing  more  than  10  1 of  culture)  with 
rDN.A  that  is  rigoi'ously  characterized  and  free 
of  harmful  seciuences;  and  3)  supporting  labora- 
tory safety  training  programs.  Every  action 
taken  by  the  Director  pursuant  to  the  Guide- 
lines must  present  "no  significant  risk  to  health 
or  the  en\  ironment.” 

R.AC  is  an  adx  isory  committee  to  the  Director 
on  technical  matters.  It  meets  quarterly.  Its  pur- 
pose, as  described  in  its  current  charter  of  June 
26,  1980  (and  unchanged  since  its  inception  in 
October  1974),  is  as  follows: 

The  goal  of  the  Committee  is  to  investigate 
the  current  state  of  knowledge  and  technology 
regarding  DNA  recombinants,  their  survival  in 
nature,  and  transferability  to  other  organisms; 
to  recommend  guidelines  for  the  conduct  of 
recombinant  DNA  experiments;  and  to  recom- 
mend programs  to  assess  the  possibility  of 
spread  of  specific  DNA  recombinants  and  the 
possible  hazards  to  public  health  and  to  the  en- 
vironment. This  Committee  is  a technical  commit- 
tee, established  to  look  at  a specific  problem.  (Em- 
phasis added.) 

The  charter  and  the  Guidelines  also  assign  it 
certain  advisory  functions  that  have  changed 
over  time. 

The  RAC  is  composed  of  not  more  than  25 
members.  At  least  eight  must  specialize  in  mo- 
lecular biology  or  related  fields;  at  least  six  must 
be  authorities  from  other  scientific  disciplines; 
and  at  least  six  must  be  authorities  on  law, 
public  policy,  the  environment,  public  or  oc- 
cupational health,  or  related  fields.  In  addition. 


214  • Impacts  of  Applied  Genetics— Micro-Organisms,  Piants,  and  Animais 


representatives  from  various  Federal  agencies 
serve  as  nonvoting  members. 

ORDA  performs  administrative  functions, 
which  include  reviewing  and  approving  IBC 
membership  and  serving  as  a national  center 
for  information  and  advice  on  the  Guidelines 
and  rDNA  activities. 

The  Interagency  Committee  was  established  in 
October  1976  to  advise  the  Secretary  of  the  then 
Department  of  Health  Education  and  Welfare 
(HEW)  [now  Health  and  Human  Services 
(DHHS)]  and  the  Director  of  NIH  on  the  coor- 
dination of  all  Federal  activities  relating  to 
rDNA.  It  has  thus  far  produced  two  reports.  Its 
first,  in  March  1977,  concluded  that  existing 
Federal  law  would  not  permit  the  regulation  of 
all  rDNA  research  in  the  United  States  to  the  ex- 
tent considered  necessary®  and  recommended 
new  legislation,  specifying  the  elements  of  that 
legislation.'®  The  second,  in  November  1977, 
surveyed  international  activities  on  regulating 
the  research  and  concluded  that,  while  appro- 
priate Federal  agencies  should  continue  to  work 
closely  with  the  various  international  organiza- 
tions, no  formal  governmental  action  was  neces- 
sary to  produce  international  control  by  means 
of  a treaty  or  convention."  It  is  currently  con- 
sidering issues  arising  from  the  large-scale  in- 
dustrial applications  of  rDNA  techniques. 

Under  the  Guidelines,  essentially  all  the  re- 
sponsibility for  overseeing  rDNA  experiments 
lies  with  those  sponsoring  or  conducting  the  re- 
search. The  Institution  must  implement  general 
safety  policies,*  establish  an  IBC,  which  meets 
specified  requirements,  and  appoint  a Biological 
Safety  Officer.  The  Biological  Safety  Officer,  who 
is  needed  only  if  the  Institution  conducts  ex- 
periments requiring  P3  or  P4  containment,  (see 
table  35)  oversees  safety  standards.  The  initial 
responsibility  for  particular  experiments  lies 


^Interim  Report  of  the  Federal  Interagency  Committee  on  Recom- 
binant DNA  Research:  Suggested  Elements  for  Legislation,  Mar.  15, 
1977,  pp.  9-10. 

'“Ibid.,  pp.  1 1-15. 

"Report  of  the  Federal  Interagency  Committee  on  Recombinant 
DNA  Research:  International  Activities,  November  1977,  pp.  13-15. 

‘These  include  conducting  any  bealib  surveillance  ibat  it  deter- 
mines to  be  necessary  and  ensuring  appropriate  ti  aining  tor  the 
IBC,  Biological  Satety  Otlicers,  Principal  Investigators,  and  labora- 
tory staff. 


with  the  PI,  the  scientist  receiving  the  funding. 
This  person  is  responsible  for  determining  and 
implementing  containment  and  other  safe- 
guards and  training  and  supervising  staff.  In  ad- 
dition, the  PI  must  also  submit  a registration 
document  that  contains  information  about  the 
project  to  the  IBC,  and  petition  NIH  for:  1)  cer- 
tification of  host-vector  systems,  2)  exceptions 
or  exemptions  from  the  Guidelines,  3)  and  de- 
termination of  containment  levels  for  experi- 
ments not  covered  by  the  Guidelines.  Further- 
more, all  of  the  above  have  certain  reporting  re- 
quirements designed  so  that  ORDA  is  eventually 
informed  of  significant  problems,  accidents,  \ io- 
lations,  or  illnesses.  * * 

The  IBC  is  designed  to  prox  ide  a (|uasi-inde- 
pendent  review  of  rDNA  work  done  at  an  in- 
stitution. It  is  responsible  for:  1)  rex  iewing  all 
rDNA  research  conducted  at  or  s[)onsored  by 
the  institution  and  approxing  those  pi'ojects  in 
conformity  with  the  Guidelines:  2)  periodically 
reviewing  ongoing  projects;  3)  adopting  emer- 
gency plans  for  spills  and  contamination;  4) 
lowering  containment  levels  for  certain  rDN.A 
and  recombinant  organisms  in  xvhich  th(>  ab- 
sence of  harmful  se(|uences  has  hec'ii  (>stah- 
lished;  and  5)  reporting  significant  problems, 
violations,  illnesses,  or  accidents  to  ORD.A 
within  30  days.***  Fhe  IBC!  must  he  com|)rised 
of  no  fewer  than  five  members  xxho  can  col- 
lectively assess  the  risks  to  health  or  the  en- 
vironment from  the  (bxperiments.  At  least  20 
percent  of  the  memh(M\ship  must  not  he  other- 
wise affiliated  with  the  institution  xxhere  the 
work  is  being  done,  and  must  re[)re.sent  the  in- 
terests of  the  surrounding  community  in  jiro- 
tecting  health  and  th(?  enx  ironment.  Comm- 
mittee  members  cannot  rexiexx  a project  in 
which  they  hax  e been,  or  e,\|)ect  to  he,  inxoix  ed 
or  have  a direct  finant'ial  interest.  Finally,  ilu' 
Guidelines  suggest  that  IBC  meetings  he  public: 
minutes  of  the  m(u4ings  and  submitted  docu- 
ments must  h(!  axailahle  to  the  public  on 
request. 


* ‘ The  I’l  Ls  rc(|uit'(‘(l  to  rcporl  this  inliM'm.iliiin  ilhin  .1(1  d.n  -•  In 
ORDA  and  bi.s  IB(  . I he  Biological  Salcl\  Olliccc  iiiiinI  ll•(Mll  l (hi- 
same  lo  llui  Inslidilion  and  ibe  IB(  unless  the  1*1  has  done  so  I be 

Inslitullon  nuisl  reporl  uilbln  .III  d.i\s  lo  ()RI)\  unless  ihe  I'l 
IBC  has  done  so. 

“'ll  does  nol  ba\i‘  lo  reporl  il  llie  I’l  h.is  done  so 


Ch.11 — Regulation  of  Genetic  Engineering  • 215 


rhe  reciuirements  imposed  on  an  institution 
and  its  scientists  are  enforced  l)v  the  authority 
of  N'lH  to  suspend,  terminate,  or  place  other 
conditions  on  its  funding  of  the  offending  proj- 
ects or  all  projects  at  the  institution.  Compliance 
is  monitored  through  the  requirements  for  noti- 
fication mentioned  aho\  e. 

PROVISIONS  FOR  \()Ll\T.ARV  CO.MPLI ANCE 

Organizations  or  indi\  iduals  w ho  do  not  re- 
cei\e  any  \'IH  funds  for  rUN'.A  research  are  not 
coxered  hy  the  Cuidelines.  These  include  other 
Federal  agencies,  institutions  and  indixiduals 
funded  by  those  agencies,  and  corporations. 

Federal  agencies  other  than  i\'IH  that  conduct 
or  fund  rD\,\  research  ha\e  proclaimed  their 
\oluntary  compliance  with  the  Guidelines.* 
Staff  scientists  ha\  e been  so  informed  hy  memo- 
randa. .As  foi'  outside  inxestigators,  this  policy 
has  been  implemented  through  the  grant  appli- 
cation process.  Instructions  in  grants  appli- 
cations contain  policy  statements  regarding 
compliance  w ith  the  Guidelines,  and  applicants 
are  sometimes  contacted  to  ascertain  their 
knowledge  of  the  Guidelines.  Information  has 
been  requested  for  certain  e.xperiments,  and 
IBC  membership  has  been  rex  iewed.  From  time 
to  time,  the  agencies  haxe  consulted  xxith  NIH 
on  matters  that  need  interpretation. 

Part  \ I of  the  Guidelines  is  designed  to  en- 
courage xoluntary  compliance  by  industry.  It 
creates  a parallel  system  of  project  reviexv  and 
IBC  approxal  analogous  to  that  required  for 
\IH-funded  projects,  modified  to  allex  iate  in- 
dustry’s concerns  about  protection  of  pro- 
prietary information. 

The  Freedom  of  Information  Act  requires 
Federal  agencies,  xxith  certain  exceptions,  to 
make  their  records  ax  ailable  to  the  public  on  re- 
quest. One  of  the  exceptions  is  for  trade  secrets 
and  proprietary  information  obtained  from 
others.  Part  \'I  contains  sexeral  provisions  for 
protecting  this  information.  Perhaps  the  most 
important  is  a process  xvhereby  a corporation 

•These  agencies  are  the  National  Science  Foundation,  the  De- 
partment of  Agriculture,  the  Department  of  Energ\',  the  X eterans 
•Administration,  and  the  Center  for  Disease  Control.  Two  other 
agencies,  which  have  e.xpressed  interest  in  this  research  but  are 
not  currently  sponsoring  any  projects,  are  the  Department  of  De- 
fense and  the  National  .Aeronautics  and  Space  .Administration. 


may  request  a presubmission  reviexv  of  the 
records  needed  to  register  its  projects  xvith  NIH. 
The  DHHS  Freedom  of  Information  Officer 
makes  an  informal  determination  of  whether 
the  records  xvould  haxe  to  he  released.  If  they 
are  determined  to  be  releasable,  the  records  are 
returned  to  the  submitting  company.  The 
Guidelines  also  require  that  NIH  consult  xvith 
any  institution  applying  for  an  exemption, 
exception,  or  other  approx  al  about  tbe  content 
of  any  public  notice  to  be  issued  xvben  the  ap- 
plication inx  olx  es  proprietary  information.  As  a 
matter  of  practice,  such  applications  are  also 
considered  by  RAC  in  nonpublic  sessions. 

Large-scale  experiments  (more  than  10  1 of 
culture)  xvith  rDNA  molecules  are  prohibited 
unless  the  rDNA  is  "rigorously  characterized 
and  the  absence  of  harmful  sequences  estab- 
lished.” Such  experiments  are  actually  scale-ups 
of  potential  industrial  processes.  Those  meeting 
this  standard  may  be  approved  by  the  Director 
of  NIH  under  procedures  specified  by  RAC.*  At 
its  September  1979  meeting,  RAC  adopted  pro- 
cedures for  reviexv  that  require  the  applicant  to 
submit  information  on  its  laboratory  practices 
and  containment  equipment.  Subsequently,  rec- 
ommendations xvere  developed  for  large-scale 
uses  of  organisms  containing  rDNA.  These  were 
published  in  the  Federal  Register  on  April  11, 
1980.  Besides  setting  large-scale  containment 
levels,  they  require  the  institution  to  appoint  a 
Biological  Safety  Officer  xvith  specified  duties, 
and  to  establish  a xvorker  health  surveillance 
program  for  xx^ork  requiring  P3  containment.  At 
its  September  1980  meeting,  RAC  modified  its 
reviexv  procedures  so  that  the  application  need 
only  specify  the  large-scale  containment  level  at 
which  the  work  xvould  be  done,  without  pro- 
viding details  on  containment  equipment.  RAC 
xvill  continue  to  review  the  biological  aspects  of 
the  applications  in  order  to  determine  that 
rDNA  is  rigorously  characterized,  that  the  ab- 
sence of  harmful  sequences  is  established,  and 
that  the  proposed  containment  is  at  the  ap- 
propriate level. 

•It  is  NIH,  not  the  company  proposing  the  scale-up,  that  deter- 
mines if  the  rDNA  to  he  used  is  "rigorously  characterized  and  the 
absence  of  harmful  sequences  established.".'^ 

‘^Guidelines  for  Research  Involving  Recombinant  DNA  Mole- 
cules, sec.  IX'-E-l-b-(3)-(d). 


216  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


Evaluation  of  the  Guidelines 

Two  basic  issues  must  be  addressed.  The  first 
is  how  well  the  Guidelines  confront  the  risks 
from  genetic  engineering,  which  may  not  have  a 
definitive  answer  in  view  of  the  uncertainty 
associated  with  most  of  the  risks.  Consequently, 
it  is  also  necessary  to  consider  a second  issue— 
whether  confidence  is  warranted  in  the  deci- 
sionmaking process  responsible  for  the  Guide- 
lines. 

THE  PROBLEM  OF  RISK 

The  Guidelines  are  designed  to  address  the 
risks  to  public  health  and  the  environment  from 
either  rDNA  molecules  or  organisms  and  vi- 
ruses containing  them.  The  underlying  premise 
is  that  research  should  not  be  unreasonably 
restricted.  This  is  essentially  a risk-benefit  ap- 
proach; at  the  time  that  the  original  Guidelines 
were  drafted,  it  represented  a compromise  be- 
tween the  extremes  of  no  regulation  and  of  no 
research  without  proof  of  safety.  Physical  and 
biological  containment  levels  were  established 
for  various  experiments  based  on  estimated 
degrees  of  risk.  The  administrative  mechanism 
created  by  the  Guidelines  is  that  of  a Federal 
agency— NIH— advised  by  a diverse  body  of 
experts— RAC.  Scientific  advice  on  the  technical 
aspects  of  risk  assessment  is  provided  by  techni- 
cal experts  on  RAC;  public  input  is  provided  by 
experts  in  nontechnical  subjects  and  by  the 
right  of  the  public  to  comment  on  major  actions, 
which  are  published  in  the  Federal  Register. 
Compliance  is  accomplished  by  a combination 
of  local  self-regulation  and  limited  Federal  over- 
sight, with  the  ultimate  enforcement  resting  in 
the  Federal  funding  power. 

Since  their  initial  appearance,  the  Guidelines 
have  evolved.  As  scientists  learned  more  about 
rDNA  and  molecular  genetics,  two  trends  oc- 
curred. First,  containment  levels  were  progres- 
sively lowered.  Major  revisions  were  made  in 
1978  and  1980;  minor  revisions  were  often 
made  quarterly,  as  proposals  were  submitted  to 
the  RAC  at  its  quarterly  meetings,  recom- 
mended by  RAC,  and  accepted  by  the  Director. 
By  now,  approximately  85  percent  of  the  per- 
mitted experiments  can  be  done  at  the  lowest 
physical  and  biological  containment  levels.  Se- 
cond, the  degree  of  centralized  Federal  over- 


sight has  been  substantially  reduced  to  the  point 
where  almost  none  remains.  Under  the  1976 
Guidelines,  all  permitted  experiments  ultimately 
had  to  be  reviewed  by  the  IBC  and  ORDA  before 
they  could  be  started;  the  1978  Guidelines  no 
longer  required  preinitiation  review  of  most 
experiments  by  ORDA,  although  ORDA  con- 
tinued to  maintain  a registry  of  experiments 
and  to  review  IBC  decisions.  Under  the 
November  1980  revision  to  the  Guidelines,  there 
will  be  no  Federal  registration  or  review  of  ex- 
periments for  which  containment  le\els  are 
specified  in  the  Guidelines.  About  97  percent  of 
the  permitted  experiments  fall  into  this 
category. 

Preinitiation  review  of  experiments  by  RA(’ 
has  been  an  important  part  of  the  oversight 
mechanism.  Expert  review  encourages  experi- 
mental design  to  be  well  thought  out  and  pro- 
vides a means  for  catching  potential  pi’ohlems, 
e.g.,  one  application  re\iewed  by  B,\(;  ne\('r 
mentioned  that  the  s[)ecies  to  he  used  as  a DN.\ 
donor  was  capable  of  manufacturing  a potent 
neurotoxin;  it  was  turned  down  aft(M'  a RAC 
member  familiar  with  the  species  brought  this 
fact  to  the  Committee’s  attention.'-’ 

The  burdens  imposed  on  rDN.A  acti\  ities  by 
the  Guidelines  appear  to  he  reasonable  in  \ iew 
of  continuing  concerns  about  risk.  I,(‘ss  than  15 
percent  of  permitted  expcM'iments  re(|uii’('  pre- 
initiation appro\  al  by  the  local  IBC's,  which  usu- 
ally meet  monthly.  Preinitiation  approx  al  of  e.\- 
periments  by  NIH  is  retjuired  only  for:  I)  e.xperi- 
ments  that  have  not  been  assigned  containment 
levels  by  the  Guidelines;  2)  expei-iments  using 
new  host-vector  systems,  which  must  he  cei  ti- 
fied  by  NIH;  3)  certain  experiments  re(|uiring 
case-by-case  approval;  and  4)  i'(*{|uests  for  ex- 
ceptions from  Guideline  re(|uii'ements.  1 he  low- 
est containment  levels  place  minimal  burdens 
on  the  experimenter,  (see  table  35).  For  in- 
dustrial applications,  NIH  approval  must  hi- 
received  not  only  when  th(‘  pi’oje('t  is  .scaled-u|) 
beyond  the  10-1  limit,  hut  also  for  each  addi- 
tional scale-up  of  the  same  project.  Many  re|)ia‘- 
sentatives  of  industry  consider  these  suhse- 


’’R.  M.  tlenig.  "Irmihic  on  llu-  H V(  ( omniitti'r  S()lii>  (hi-i 
Downgrading  of  £.  ro/i  Cont.iinnicnt.  HioSrirntr.  \n\  p|i 
762,  December  1979. 


Ch.  11— Regulation  of  Genetic  Engineering  • 217 


quent  appro\  als  to  be  unnecessary  and  burden- 
some. 

Information  about  whether  the  Guidelines 
ha\  e been  a disad\  antage  for  L'.S.  companies  in 
international  competition  is  scanty.  E.xamples 
include  the  appro.ximately  1-year  headstart  two 
European  groups  were  gi\  en  while  the  cloning 
of  hepatitis  B \ irus  was  prohibited,  the  ad\  an- 
tage  some  European  companies  had  in  using 
certain  species  of  bacteria  for  cloning  under 
conditions  that  were  prohibited  in  the  L^nited 
States,  and  the  delays  some  pharmaceutical 
companies  faced  because  they  had  to  build  bet- 
ter containment  facilities. 

The  present  (iuidelines  are  a comprehensive, 
dexible,  and  nonhurdensome  way  of  dealing 
w ith  the  physical  risks  associated  v\  ith  rDN'A  re- 
search while  permitting  the  work  to  go  for- 
ward. That  is  all  they  u ere  e\  er  intended  to  do. 

The  Scope  of  the  Guidelines.— In  many 
respects,  the  Guidelines  do  not  address  the  full 
scope  of  the  risks  of  genetic  engineering.  They 
co\  er  one  technique,  albeit  the  most  important; 
they  do  not  address  the  admittedly  uncertain, 
long-term  cultural  risks:  they  are  not  legally 
binding  on  researchers  recei\  ing  funds  from 
agencies  other  than  N’lH;  and  they  are  not  bind- 
ing on  industry. 

Other  genetic  techniques  present  risks  simi- 
lar to  those  posed  by  rD\,A,  but  to  a lesser  de- 
gree. Recombinant  Di\A  is  the  most  \ersatile 
and  efficient  technique;  it  uses  the  greatest 
\ariety  of  genetic  material  from  the  widest 
number  of  sources  with  reasonable  assurance 
of  expression  by  the  host  cell.  Cell  fusion  of 
micro-organisms,  which  also  in\  ol\  es  the  uncer- 
tain risk  of  recombining  the  genetic  material  of 
different  species,  is  significantly  less  versatile 
and  efficient  than  rDNA  but  mixes  more  genetic 
material.  In  addition,  the  parental  cells  may  con- 
tain partial  viral  genomes  that  could  combine  to 
form  a complete  genome  when  the  cells  are 
fused.  Transformation,  a technique  known  for 
decades,  similarly  imolves  moving  pieces  of 
D\A  betw  een  different  cells.  How'ever,  it  is  sig- 
nificantly less  versatile  and  efficient  than  cell  fu- 
sion, and  it  is  generally  considered  to  be  virtual- 
ly risk-free.  Thus,  cell  fusion  is  in  a gray  area 


between  the  other  two  techniques;  yet  no  risk 
assessment  has  been  done,  and  no  Federal  over- 
sight exists. 

Another  limitation  in  the  scope  of  the  guide- 
lines—and  in  the  process  by  which  they  were 
formulated— is  that  long-range  cultural  risks  (as 
distinguished  from  policy  issues  related  to  safe- 
ty) were  never  addressed.  As  noted  by  the  Di- 
rector of  NIH:'-* 

. . . NIH  has  been  addressing  the  policy  ques- 
tions in\'olving  the  safety  of  this  research,  not 
the  potential  future  application  ...  to  the  alter- 
ing of  the  genetic  character  of  higher  forms  of 
life,  including  man’ . . . 

Perhaps  it  was  inappropriate  to  do  more.  Such 
ethical  issues  might  be  considered  premature  in 
view  of  the  level  of  the  development  of  the  tech- 
nology'. The  desire  among  many  molecular  bi- 
ologists to  mo\  e ahead  w'ith  the  research  meant 
that  experiments  were  being  done;  therefore 
the  immediate  potential  for  harm  was  to  health 
and  the  en\ironment.  Thus,  it  was  arguably 
necessary  to  develop  a framework  to  deal  with 
the  risks  based  on  what  was  known  at  the  time. 
On  the  other  hand,  the  broader  questions  of 
where  the  research  might  eventually  lead  and 
whether  it  should  be  done  at  all  have  been 
raised  in  the  public  debate.  They  have  not  been 
formally  considered  by  the  Federal  Govern- 
ment. 

Another  limitation  in  the  scope  of  the  Guide- 
lines is  their  nonapplicability  to  research 
funded  or  performed  by  other  Federal  agencies. 
However,  agencies  supporting  such  research 
are  complying  with  the  Guidelines  as  a matter  of 
policy.  There  appears  to  be  little  reason  for 
questioning  these  declarations  of  general  policy. 
In  practice,  problems  might  arise  if  a mission  is 
perceived  to  be  at  odds  with  the  Guidelines  or 
because  of  simple  bureaucratic  defense  of  terri- 
tory—e.g.,  when  the  1976  Guidelines  were  pro- 
mulgated, tw'o  agencies— the  Department  of  De- 
fense (DOD)  and  the  National  Science  Founda- 
tion (NSF)— reserved  the  right  to  deviate  for  rea- 
sons of  national  security  or  differing  interpreta- 


'MS  F.R.  60103,  Dec.  22,  1978,  citing  43  F.R.  33067,  July  28, 
1978. 


218  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


tions,  respectively.*  DOD  no  longer  claims  an 
exception  for  national  security. NSF  took  its 
position  when  it  approved  funding  for  an  ex- 
periment using  a particular  species  of  yeast  that 
had  not  been  certified  by  NIH,  relying  on  an  am- 
biguously worded  section’®  in  the  Guidelines  to 
assert  that  it  could  certify  the  host.  Subsequent 
revisions  explicitly  stated  that  these  hosts  had  to 
be  certified  by  the  Director  of  NIH”"  and 
removed  many  similar  ambiguities. 

In  the  final  analysis,  NIH  has  indirect  leverage 
over  the  actions  of  other  agencies  through  its 
funding.  All  non-NIH  funded  rDNA  projects  at 
an  institution  which  also  receives  NIH  funds  for 
rDNA  work  must  comply  with  the  Guidelines; 
otherwise  NIH  funds  may  be  suspended  or  ter- 
minated. 

While  the  procedures  of  other  agencies  for 
administering  compliance  are  significantly  less 
formal  than  those  created  by  the  Guidelines  for 
NIH,  they  do  rely  heavily  on  NIH  for  help  and 
advice,  and  they  coordinate  their  efforts 
through  the  Interagency  Committee  and  their 
nonvoting  membership  on  RAC.  So  far,  this  vol- 
untary compliance  by  the  agencies  appears  to 
be  working  fairly  well. 

The  most  significant  limitation  in  the  scope  of 
the  Guidelines  is  their  nonapplicability  to  in- 
dustrial research  or  production  on  other  than  a 
voluntary  basis.  This  lack  of  legal  authority 
raises  concerns  not  only  about  compliance  but 
also  about  NIH’s  ability  to  implement  a volun- 
tary program  effectively. 

Whether  every  company  working  with  rDNA 
will  view  voluntary  compliance  to  be  in  its  best 
interest  depends  on  a number  of  factors.  In 
the  past,  certain  short-sighted  actions  by  even  a 
few  companies  in  a given  industry  has  led  to 

*For  a statement  of  the  DOD  position,  see  the  minutes  of  the 
November  23,  1976,  meeting  of  the  Federal  Interagency  Commit- 
tee. At  that  time,  DOD  had  no  active  or  planned  rDNA  projects. 
NSF’s  statement  of  its  intention  to  "preserve  some  level  of  inde- 
pendence of  decision"  was  expressed  in  an  internal  NIH  memo- 
randum dated  February  24,  1978,  from  the  Deputy  Director  for 
Science,  NIH,  to  the  Director,  NIH. 

'®Dr.  John  H.  Moxley,  III.,  Assistant  Secretary  of  Defense  for 
Health  Affairs,  personal  communication,  Nov.  18,  1980, 

''"Fungal  or  Similar  Lower  Eukaryotic  Host- Vector  Systems,"  41 
F.R.  27902,  27920,  July  7,  1976. 

'^43  F.R.  60108,  Dec.  22,  1978,  sec.  IIl-C-5  of  the  1978  Guidelines. 

■MS  F.R.  6724,  Jan.  29,  1980,  sec.  III-C-5  of  the  1980  Guidelines. 


well-documented  abuses  and  a host  of  Federal 
laws  to  curtail  them.  However,  at  least  two  con- 
straints are  operating  in  the  case  of  the  bio- 
technology industry.  First,  the  possibility  of  tort 
lawsuits  is  an  inducement  to  comply  with  the 
Guidelines,  which  would  probably  be  accepted 
as  the  standard  of  care  against  which  alleged 
negligence  would  be  evaluated.  (This  concept  is 
discussed  in  greater  detail  in  the  section  on  Tort 
Law  and  Workman’s  Compensation.)  Second, 
the  threat  of  statutory  regulation,  which  the 
companies  have  sought  to  avoid,  always  exists. 
Other  factors  are  also  at  work.  Except  for  the 
10-1  limitation,  for  which  case>by-case  excep- 
tions must  be  sought,  the  large-scale  contain- 
ment recommendations  of  April  11,  1980,  are 
not  excessively  burdensome,  at  least  for  phar- 
maceutical companies.  The  requirements  are 
similar  to  measures  that  must  currently  he 
taken  to  prevent  product  contamination.  In  ad- 
dition, the  public  debate  should  have  made  each 
company  aware  of  the  problems  and  the  need 
for  voluntary  compliance  before  it  inv  ested  sub- 
stantially in  biotechnology;  expensive  controls 
will  not  have  to  be  retrofitted.  However,  one 
definite  concern  is  that  new  com|)anies  at- 
tracted to  the  field  will  perceive  their  interests 
differently.  Because  they  did  not  actually  expe- 
rience the  period  when  legislation  seemed  ine\  i- 
table  and  because  they  will  he  late  entries  in  th(> 
race,  they  may  be  inclined  to  take  shortcuts. 

Besides  the  concern  about  whether  iiulustry 
has  sufficient  incentive  to  comply,  theri’  ar(>  a 
number  of  other  reasons  for  (luestioning  th(>  ef- 
fectiveness of  the  voluntary  program.  First, until 
very  recently  no  member  of  KAt!  was  an  e,\|)ert 
in  industrial  fermentation  technology— yet  the 
Committee  has  been  considering  applications 
from  industry  for  large-scale  production  since 
September  1979.*  This  drawback  was  demon- 
strated at  its  March  1980  meeting,  when  the 
Committee  expressed  uncertainty  owr  what 
Federal  or  State  safety  regulations  [iresently 
cover  standard  fermentation  technolog^v  I’ln- 

‘At  its  September  1980  meeting.  R \(  p.issed  the  InllmMiig  res 
oliUion,  wliieb  hits  been  accepted  b\  the  Du  et  tnr  ol  \IM  ” 

\teml)ci‘.s  should  he  chosen  to  pi'oude  expei  lise  in  lei  inent.ilinn 

lechnologv,  (engineering,  .ini  1 oilier  ,is|iec1s  i it  l.n  ge  si  .ile  jn  inIih  In  >n 

A termentation  lechnnlng\  expert  w.is  appointed  m l.iim.iix 
1981. 

'M.l  F.R.  77373,  Nov.  21,  Iil8l) 


Ch.  11— Regulation  of  Genetic  Engineering  • 219 


ployed  by  the  drug  industry.  \ arious  members 
e.xpressed  concern  in  the  March  and  June  1980 
meetings  about  the  Committee’s  continuance  to 
make  recommendations  on  the  applications 
without  a firm  knowledge  of  large-scale  produc- 
tion. 

Second,  the  pro\  isions  in  part  \ I of  the  Guide- 
lines, which  allow  prior  re\  iew  of  submitted  in- 
formation by  the  DHHS  Freedom  of  Information 
.Act  Officer,  gi\  e an  industrial  applicant  the  op- 
tion of  withholding  potentially  important  infor- 
mation on  the  grounds  of  trade  secrecy,  e\en 
when  DHHS  disagrees.  Third,  because  some 
R AC  members  ha\  e been  opposed  to  discussing 
industrial  applications  in  closed  session  (needed 
to  protect  proprietary  information),  they  have 
chosen  not  to  participate  in  those  sessions. 
Thus,  some  di\ersity  of  opinion  and  e.xpertise 
has  been  lost.  Fourth,  monitoring  for  compli- 
ance after  the  scale-up  applications  are  granted 
is  limited.  Some  early  applications  were  granted 
on  the  condition  that  \1H  could  inspect  facili- 
ties, and  at  least  one  inspection  was  made. 
Under  procedures  adopted  at  the  September 
1980  meeting,  a company’s  IBC  will  be  responsi- 
ble for  determining  whether  the  facilities  meet 
the  standards  for  the  large-scale  containment 
level  assigned  by  R.AC.  A working  group  of  RAC 
may  visit  the  companies  and  their  IBCs  from 
time-to-time  but  only  for  information  gathering 
purposes,  rather  than  for  regulatory  actions. 
Fifth,  even  if  noncompliance  were  found,  no 
penalties  can  be  imposed. 

The  members  of  R.AC,  acutely  aware  of  the 
problems  with  voluntary  compliance  by  indus- 
try, ha\e  been  deliberating  about  them  for 
almost  2 years.  At  a meeting  in  May  1979,  they 
decided,  by  a vote  of  nine  to  six  with  six  absten- 
tions, to  support  the  principle  of  mandatory 
compliance  with  the  Guidelines  by  non-MH 
funded  institutions.  However,  the  Secretary  of 
HEW'  (Joseph  Califano)  decided  to  continue  with 
the  dex  elopment  of  \ oluntary  compliance  provi- 
sions^® which  were  adopted  as  Part  \'I  of  the 
Guidelines  in  January  1980.  Actual  RAC  review 
of  submissions  from  the  private  sector  for  large- 
scale  work  began  in  September  1979.  At  a meet- 
ing in  June  1980,  RAC  debated  the  effectiveness 

“R.AC  minutes  of  Sept.  6-7,  1979,  p.  16,  in  Recombinant  DXA  Re- 
search, vol.  5,  (Wash.,  D.C.:  HEW,  1980),  p.  165. 


of  NlH’s  quasi-regulation  of  industry.  A primary 
concern  was  whether  the  RAC  would  be  viewed 
as  gix'ing  a “stamp  of  approval”  to  industrial  pro- 
jects, when,  in  fact,  it  has  neither  the  authority 
nor  the  ability  to  do  so.  One  member,  lawyer 
Patricia  King,  stated:^* 

Voluntary  compliance  is  the  worst  of  all  possi- 
ble worlds  . . . .You  achieve  none  of  the  objec- 
ti\es  of  regulation  and  none  of  the  benefits  of 
being  unregulated.  All  you’re  saying  is  'I  give  a 
stamp  of  approval  to  what  1 see  here  before  me 
without  any  authority  to  do  anything.’ 

Most  of  the  speakers  expressed  the  desire  that 
the  \ arious  agencies  in  the  Interagency  Commit- 
tee be  responsible  for  such  regulation.  How- 
ever, the  Interagency  Committee,  which  has 
been  studying  the  problem  since  January  1980, 
has  yet  to  decide  what  it  can  do.  Thus,  many  of 
its  members  see  RAC  as  filling  a regulatory  void 
until  the  traditional  agencies  take  action. 

Some  regulatory  agencies  have  begun  to  deal 
with  specific  problems  within  their  areas  of  in- 
terest. The  Occupational  Safety  and  Health  Ad- 
ministration will  decide  its  regulatory  policy  on 
the  basis  of  a study  of  potential  risks  to  workers 
posed  by  the  industrial  use  of  rDNA  techniques 
being  conducted  by  the  National  Institute  of 
Occupational  Safety  and  Health  (NIOSH).  In  a 
letter  to  the  Director  of  NIH  dated  September 
24,  1980,  Dr.  Eula  Bingham,  then  Assistant  Sec- 
retary for  Occupational  Safety  and  Health  of  the 
Department  of  Labor,  estimated  this  process 
would  take  approximately  2 years.  The  Environ- 
mental Protection  Agency  (EPA)  has  awarded 
several  contracts  and  grants  to  assess  the  risks 
of  intentional  release  of  genetically  engineered 
micro-organisms  and  plants  into  the  environ- 
ment. And  the  Food  and  Drug  Administration 
(FDA)  has  begun  to  develop  policy  with  respect 
to  products  made  by  processes  using  genetically 
engineered  micro-organisms.  (Further  details 
on  agency  actions  are  discussed  in  the  section. 
Federal  Statutes.) 

Compliance.— The  primary  mechanism  in 
the  Guidelines  for  enforcing  compliance  is  local 
self -regulation,  with  very  limited  Federal  over- 


^'Susan  Wright,  'Recombinant  DNA  Policy:  Controlling  Large- 
Scale  Processing,”  Environment,  vol.  22,  September  1980,  pp. 
29,32. 


220  • Impacts  of  Applied  Genetics— Micro-Organisms,  Piants,  and  Animals 


sight.  Penalties  are  based  on  NIH’s  power  to  re- 
strict or  terminate  its  funding. 

The  initial  responsibility  for  compliance  lies 
with  the  scientist  doing  the  experiments.  A re- 
searcher’s attitude  toward  the  risks  of  rDNA 
techniques  and  the  necessity  for  the  Guidelines 
appear  to  be  an  influential  factor  in  the  degree 
of  compliance.  A science  writer  who  worked  for 
3 months  in  a university  lab  in  1976  noted  slop- 
py procedures  and  a cavalier  attitude,  stating: 
“Among  the  young  graduate  students  and  post- 
doctorates it  seemed  almost  chic  not  to  know 
the  NIH  rules. On  the  other  hand,  in  the  case 
of  a recent  violation  of  the  Guidelines,  it  appears 
as  if  the  investigator’s  graduate  students  were 
the  first  to  raise  questions. Competitiveness 
is  another  important  factor.  Novice  scientists 
must  establish  reputations,  secure  tenure  in  a 
tight  job  market,  and  obtain  scarce  research 
funds;  established  researchers  still  compete  for 
grants  and  certainly  for  peer  recognition.  This 
competitive  pressure  could  provide  strong  in- 
centives to  bend  the  Guidelines;  on  the  other 
hand,  it  might  be  channeled  to  encourage  com- 
pliance if  it  is  believed  that  NIH  will  in  fact 
penalize  violations  by  restricting  or  terminating 
funding. 

The  first  level  of  actual  oversight  occurs  at 
the  institution.  An  argument  can  be  made  that 
reliance  on  the  PI  and  an  IBC  (that  might  be 
composed  mostly  of  the  Pi’s  colleagues)  provides 
too  great  an  opportunity  for  lax  enforcement  or 
coverups.  On  the  other  hand,  spreading  respon- 
sibility among  the  institution,  the  PI,  the  IBC, 
and,  in  the  case  of  more  hazardous  experi- 
ments, the  Biological  Safety  Officer  might  re- 
duce the  chance  of  violations  being  overlooked 
or  condoned.  This  responsibility  is  enhanced  by 
the  reporting  requirements  borne  by  each  of 
these  parties,  designed  so  that  ORDA  learns  of 
“significant’’  problems,  accidents,  violations,  and 
illnesses.  What  is  “significant”  is  not  defined. 

Public  involvement  at  the  local  level  acts  as  an 
additional  safeguard.  Twenty  percent  of  the 

“Janet  L.  Hopson,  "Recombinant  Lab  for  DNA  and  My  95  Days 
in  It,"  Smithsonian,  vol.  8,  June  1977,  p.  62. 

“D.  Dickson,  "Another  Violation  of  NIH  Guidelines,"  Nature  vol. 
286,  Aug.  14,  1980,  p.  649. 

“D.  Dickson,  "DNA  Recombination  Forces  Resignation,"  Nature 
vol.  287,  Sept.  18,  1980,  p.  179. 


IBCs  members  must  be  unaffiliated  with  the  in- 
stitution. IBC  documents,  including  minutes  of 
meetings,  are  publicly  available,  but  meetings 
are  not  required  to  be  held  in  public.  On  the 
other  hand,  the  probable  inability  of  the  mem- 
bers who  represent  the  public  to  understand 
the  technical  matters  might  limit  their  effective- 
ness. 

How  successful  has  compliance  been?  Three 
known  violations  have  occurred.  In  each,  no 
threat  to  health  and  the  environment  existed.  In 
each,  there  was  some  confusion  as  to  why  the 
violations  occurred.  NIH  is  presently  in\est- 
igating  the  third  violation.  For  the  first  two,  it 
accepted  explanations  of  misunderstandings 
and  misinterpretations  of  the  Guidelines.  How- 
ever, a Senate  oversight  report  concluded:*® 

While  undoubtedly  most  researchers  ha\e 

observed  the  guidelines  conscientiously,  it  is 

equally  clear  that  others  have  substituted  their 

own  judgments  of  safety  for  those  of  NIH. 

No  firm  conclusions  can  be  drawn  on  the  (jues- 
tion  of  compliance.  The  reporting  of  only  a few 
violations  could  be  evidence  that  the  compliance 
mechanism  embodied  in  tbe  Guidelines  has 
been  working  well.  Or  it  could  mean  that  some 
violations  are  not  being  discovered  or  reported. 

Tbe  November  1980  amendments  to  the 
Guidelines  substantially  cbanged  procedure's 
designed  to  monitor  compliance  by  abolishing  a 
document  called  a Memorandum  of  I'nder- 
standing  and  Agreement  (MllA).  It  had  been  re'- 
quired  for  15  to  20  percent  of  all  e'xperime'nts, 
those  thought  to  Ije  potentially  most  risk\’.  I'he* 
MUA,  which  was  to  be  filed  with  ()MI).\  by  an 
institution,  provided  information  about  each  e.\- 
periment,  and  it  was  the  institution's  certifica- 
tion to  NIH  that  the  experiment  complie'd  w ith 
the  Guidelines.  By  having  the  Ml'.Xs,  OBD.A 
could  monitor  for  inconsistencies  in  interpret- 
ing the  Guidelines,  actual  non{'om[)liance,  and 
the  consistency  and  (luality  with  which  IB(!s 
functioned  nationwide.  The  amendments  con- 
tinued a trend  begun  in  January  1!)80,  when  ap- 
proximately 80  percent  of  the  experiments. 


^’"Recombinant  DNA  Rt'.searcli  and  Its  Vppla  ations  n\rr\ifihl 
Report,  Siihcommitlee  on  Sciencr,  I cchnoliigv  and  Spati-  ol  ibe 
Senate  Committee  on  Commerce,  Science  and  1 1 anN|«>i  tation 
Aug.  1978,  p.  17. 


Ch.  11 — Regulation  of  Genetic  Engineering  • 221 


those  done  with  E.  coli  K-12,  were  exempted 
from  the  MUA  rec|uirement. 

The  aholition  of  the  Ml^A  essentially  abol- 
ished centralized  Federal  monitoring  of  rDNA 
experiments.  The  only  current  (iuideline  provi- 
sion that  ser\  es  this  kind  of  monitoring  function 
is  the  requirement  that  the  institution,  the  IBC, 
or  the  PI  notify  OKD.A  of  any  significant  \ iola- 
tions,  accidents,  or  problems  with  interpreta- 
tion. lamited  monitoi'ing  of  large-scale  acti\  ities 
continues.  Under  \'IH  procedures  (which  are 
not  part  of  the  Ciuidelines)  for  re\  iewing  appli- 
cations for  exemptions  from  the  10-1  limit,  the 
application  must  include  a copy  of  the  registra- 
tion document  filed  with  the  IBC.  Fhe  manufac- 
turing facilities  may  also  he  inspected  by  NIH, 
not  for  regulatory  purposes,  but  to  gather  infor- 
mation for  updating  its  I'ecommended  large- 
scale  containment  levels.  The  aholition  of  the 
ML’.A  is  consistent  with  traditional  views  that 
Government  should  not  interfere  with  basic  sci- 
entific research.  \\  hether  or  not  it  will  reduce 
either  the  incentive  to  comply  with  the  Guide- 
lines or  the  likelihood  of  discovering  violations 
remains  to  be  seen. 

THE  DECISIONMAKING  PROCESS 
Another  way  to  evaluate  the  Guidelines  be- 
sides considering  their  substantive  require- 
ments is  to  look  at  the  process  by  which  they 
were  formulated.  In  a situation  where  there  is 
uncertainty  and  even  strong  disagreement 
about  the  nature,  scope,  and  magnitude  of  the 
risks,  it  is  difficult  to  judge  whether  or  not  a 
proposed  solution  to  a problem  will  be  a good 
one.  Society’s  confidence  in  the  decisionmaking 
process  and  in  the  decisionmakers  then  be- 
comes the  issue.  As  David  L.  Bazelon,  Chief 
Judge  of  the  U.  S.  Court  of  Appeals  for  the  Dis- 
trict of  Columbia,  has  stated:^® 

When  the  issues  are  controversial,  any  deci- 
sion may  fail  to  satisfy  large  portions  of  the  com- 
munity. But  those  who  are  dissatisfied  with  a 
particular  decision  will  be  more  likely  to  ac- 
quiesce in  it  if  they  perceiv’e  that  their  view's  and 
interests  were  given  a fair  hearing.  If  the  deci- 
sion-maker has  frankly  laid  the  competing  con- 
siderations on  the  table,  so  that  the  public 
knows  the  worst  as  well  as  the  best,  he  is  unlike- 

L.  Bazelon.  "Coping  With  Technology  Through  the  Legal 
Process,"  62  Cornell  Law  Review  817,825,  June  1977. 


ly  to  find  himself  accused  of  high-handedness, 
deceit,  or  cover-up.  W'e  simply  cannot  afford  to 
deal  with  these  vital  issues  in  a manner  that  in- 
V ites  public  cynicism  and  distrust. 

The  manner  in  which  the  Guidelines  them- 
selves evolved  has  been  controversial.  (For  a 
detailed  discussion  see  app.  IIl-A.)  Initially,  the 
scope  and  nature  of  the  problem  was  defined  by 
the  scientific  community;  NIH  organized  RAC 
along  the  lines  suggested  by  tbe  NAS  committee 
letter  referred  to  in  app.  III-A.  One  of  the  goals 
of  RAC  was  to  recommend  guidelines  for  rDNA 
experiments;  it  was  not  charged  with  consider- 
ing broader  ethical  or  policy  issues  or  the  funda- 
mental question  of  whether  the  research  should 
have  been  permitted  at  all.  The  original  Guide- 
lines were  produced  by  a committee  having 
only  one  nonscientist. 

In  late  1978,  the  Secretary  of  HEW  signif- 
icantly restructured  RAC  and  modified  the 
Guidelines  in  order  to  increase  the  system’s 
accountability  to  the  public,  to  "provide  the  op- 
portunity for  those  concerned  to  raise  any 
ethical  issues  posed  by  recombinant  DNA  re- 
search", and  to  make  RAC  "the  principal  ad- 
visory body  ...  on  recombinant  DNA  policy. 
However,  it  has  remained  in  large  part  a tech- 
nically oriented  body.  Its  charter  was  not 
changed  in  this  respect;  the  Guidelines  them- 
selves state  that  its  advice  is  "primarily  scientific 
and  technical,”  and  matters  presented  for  its 
consideration  have  continued  to  be  mostly  tech- 
nical. One  area  where  RAC  has  played  a signifi- 
cant policy  role,  however,  is  in  dealing  with  the 
issue  of  voluntary  compliance  by  industry. 

It  could  be  argued  that  the  system  did  provide 
for  sufficient  public  input  into  the  formulation 
of  the  problem*  and  that  no  other  formulation 
was  realistic.  The  two  meetings  in  1976  and 
1977  of  the  NIH  Director’s  Advisory  Committee 
and  the  hearing  chaired  by  the  general  counsel 
of  HEW  in  the  fall  of  1978  provicfed  the  oppor- 
tunity for  public  comment  on  the  overall  Fed- 

^Uoseph  A.  Califano,  "Notice  of  Revised  Guidelines— Recombi- 
nant DNA  Research,"  43  F.R.  60080-60081,  Dec.  22,  1978. 

'The  problem  was  conceived  in  terms  of  how  to  permit  the  re- 
search to  be  done  while  limiting  the  physical  risks  to  an  acceptable 
level.  Other  formulations  were  possible,  the  broadest  being  how 
to  limit  all  risks,  including  cultural  ones,  to  an  acceptable  level. 
Such  a formulation  could  have  resulted  in  a prohibition  of  the 
research. 


222  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


eral  approach  to  the  controversy;  including 
whether  or  not  the  problem  had  been  too  nar- 
rowly phrased.  Similarly,  Congress  had  the 
opportunity  in  1977  to  reevaluate  the  entire 
institutional  response,  taking  into  account  any 
moral  objections  to  the  research  in  addition  to 
those  concerning  safety.  Yet  the  principal  bills 
were  based  on  the  proposition  that  the  research 
continue  in  a regulated  fashion. 

A related  issue  is  the  one  of  burden  of  proof. 
Should  the  proponents  of  a potentially  benefi- 
cial technology  be  required  to  demonstrate 
minimal  or  acceptable  risk  even  if  that  risk  is 
uncertain  or  even  hypothetical?  Or  should  its 
opponents  be  required  to  demonstrate  unac- 
ceptable risk?  If  the  proposition  is  accepted  that 
those  who  bear  the  risks,  in  this  case  the  public 
as  well  as  the  scientists,  must  judge  their  ac- 
ceptability, then  the  burden  must  be  on  the  pro- 
ponents. The  scientific  community  clearly  ac- 
cepted this  burden.  The  moratorium  proposed 
by  the  NAS  committee  in  July  1974  called  for  a 
suspension  of  certain  types  of  rDNA  experi- 
ments until  the  risks  could  be  evaluated  and 
procedures  for  adequately  dealing  with  those 
risks  could  be  developed.  The  Guidelines  pro- 
hibited some  experiments,  specified  contain- 
ment levels  for  others,  and  required  certifica- 
tion of  host-vector  systems.  All  actions  approved 
by  the  Director  of  NIH,  including  the  lessening 
of  the  restrictions  imposed  by  the  original 
Guidelines,  have  had  to  meet  the  requirement  of 
presenting  “no  significant  risk  to  health  or  the 
environment.” 

Two  other  criticisms  have  been  directed 
against  RAC,  particularly  in  its  early  days.  The 
first  concerned  inherent  conflicts  of  interest. 
RAC’s  members  were  drawn  from  molecular 
biology  and  related  fields.  One  of  the  early 
drafts  of  the  Guidelines  was  criticized  as  being 
“tailored  to  fit  particular  experiments  that  are 
already  on  the  drawing  boards. However, 
only  a few  of  the  members  were  actually  work- 
ing with  rDNA. A more  serious  criticism  was 
the  lack  of  a broad  range  of  expertise.  Although 


Wade,  “Recombinant  DNA:  NIH  Sets  Strict  Rules  to  Launch 
New  Technology,"  190  Science  1175,1179,  1975. 

“Dr.  Elizabeth  Kutter,  a member  of  RAC  at  that  time,  personal 
communication.  Sept.  11,  1980. 


the  risks  had  been  expressed  in  terms  of  poten- 
tial hazards  to  human  health  and  the  environ- 
ment, the  original  RAC  had  no  experts  in  the 
areas  of  epidemiology,  infectious  diseases,  bot- 
any or  plant  pathology,  or  occupational  health. 
It  did  have  one  expert  in  enteric  organisms,  E. 
coli  in  particular. 

These  shortcomings  were  eventually  rem- 
edied by  expanding  RAC’s  membership  to  allow 
the  appointment  of  other  experts,  including 
some  from  nontechnical  fields  such  as  law  and 
ethics.  In  addition  to  providing  knowledge  of 
other  fields,  these  members  served  as  disin- 
terested advisors,  since  they  had  no  direct  in- 
terest in  expediting  the  research.  Thus,  the  Gov- 
ernment dealt  with  the  problem  of  conflicts  of 
interest  by  offsetting  the  interested  group  with 
other  groups.  In  view  of  the  need  for  the  tech- 
nical expertise  of  the  molecular  biologists,  this 
approach  seems  reasonable;  nevertheless  the 
matter  could  probably  have  been  handled  more 
expeditiously.  Although  the  April  1975  amend- 
ment to  the  RAC  charter  added  experts  from 
such  fields  as  epidemiology  and  infectious  dis- 
eases, the  charter  did  not  reciuire  plant  expi’i'ts 
until  September  1976  (shortly  aftei’  the  passage 
of  the  original  Guidelines)  and  occupational 
health  specialists  until  December  1978.  In  addi- 
tion, while  two  nontechnical  members  wen?  ad- 
ded in  1976  (one  before  and  one  aftei’  passage  of 
the  Guidelines),  their  number  was  not  inci’eased 
until  Secretary  Califano  reconstituted  the  t'om- 
mittee  in  late  1978. 

The  present  makeup  of  RAC  is  fairly  diverse. 
As  of  September  1980,  nine  of  its  members  sj)e?- 
cialized  in  molecular  biology  or  related  fields, 
seven  were  from  other  scientific  disciplines, 
and  eight  were  from  the  areas  of  law  , public 
policy,  the  environment,  and  public  or  occupa- 
tional health. 23  Moreover,  since  D(?cemher  1978, 
representatives  of  the  interested  Federal  agen- 
cies have  been  sitting  as  nonvoting  members.  In 
January  1981,  an  expert  on  fermentation  was 
added. 


^“Dr.  Bernard  Talhol,  S|)ecial  Assislant  to  Ihi-  nii  erim  MM  (wr 
sonal  comunication,  .Sept.  18.  198(1 


Ch.11 — Regulation  of  Genetic  Engineering  • 223 


One  conflict  of  interest  not  soh  ed  by  expand- 
ing the  dix  ersity  of  the  RAC’s  membership  is  in- 
stitutional in  nature.  the  agency  hax  ing  pri- 
mary responsibility  for  developing  and  adminis- 
tering the  Guidelines,  \ ie\vs  its  mission  as  one  of 
promoting  biomedical  research.  Although  the 
Guidelines  are  not  regulations,  they  contain 
many  of  the  elements  of  regulations.  They  set 
standards,  offer  a limited  means  to  monitor  for 
compliance,  and  proxide  for  enforcement,  at 
least  for  institutions  receix  ing  NIH  grants  to  do 
rDN'.A  xvork;  thus,  they  may  be  considered 
quasi-regulatory.  Regulation  is  not  only  foreign 
but  antithetical  to  XIH's  mission.  The  current 
Director  stated  publicly  at  the  June  1980  RAC 
meeting  that  the  role  of  \IH  is  not  one  of  a 
regulator,  a role  that  must  he  axoided.  Under 
these  circumstances,  perhaps  another  agency, 
or  another  part  of  DHHS,  might  he  more  appro- 
priate for  oxerseeing  the  Guidelines,  since  the 
attitudes  and  priorities  of  promoters  are  usually 
quite  different  from  that  of  regulators. 

If  R.AC  has  alxvays  been  essentially  a technical 
adx  isory  body,  xvho  then  has  made  the  x alue  de- 
cisions concerning  the  acceptability  of  the  risks 
presented  by  rDXA  and  the  means  for  dealing 
xvith  them?  The  final  decisionmaker  has  been 
the  Director  of  XIH,  xvith  the  notable  exception 
in  the  case  of  the  1978  Guidelines,  xvhich  con- 
tained the  significant  procedural  revisions 
needed  to  meet  Secretary  Califano’s  approval. 
The  Director  did  hax  e access  to  diverse  points 
of  x’iexv  through  the  Director’s  Adx  isory  Com- 
mittee meetings  and  the  public  hearings  held 
before  the  1978  Guidelines.  (See  app.  III-A.)  In 
addition,  major  actions  xvere  alxvays  accom- 
panied by  a statement  discussing  the  relevant 
issues  and  explaining  the  basis  for  the  decisions; 
after  the  1978  revisions,  major  actions  had  to  be 
proposed  for  public  comment  before  decisions 
xvere  made.  In  theory,  it  may  have  been  prefer- 
able for  the  public  to  hax  e been  substantially  in- 
volxed  in  the  actual  formation  of  the  original 
Guidelines  rather  than  simply  to  have  reacted  to 
a finished  product.  However,  this  probably 
w ould  have  sloxved  the  process  at  a time  when 
the  strong  desire  of  the  molecular  biologists  to 


^'Califano,  op.  cit. 


use  the  rDNA  techniques  could  have  threatened 
the  notion  of  self-regulation.  Today,  there  ap- 
pears to  be  reasonable  opportunity  for  public 
input  through  the  process  of  commenting  on 
proposed  actions. 

Conclusion 

The  Guidelines  are  the  result  of  an  extraor- 
dinary, conscientious  effort  by  a combination  of 
scientists,  the  public,  and  the  Federal  Govern- 
ment, all  operating  in  an  unfamiliar  realm.  They 
appear  to  be  a reasonable  solution  to  the  prob- 
lem of  hoxv  to  minimize  the  risks  to  health  and 
the  environment  posed  by  rDNA  research  in  an 
academic  setting,  xvhile  permitting  as  much  of 
that  research  as  possible  to  proceed.  They  do 
not  in  any  xvay  deal  xvith  other  molecular  genet- 
ic techniques  or  xvith  the  long-term  social  or 
philosophical  issues  that  may  be  associated  with 
genetic  engineering. 

The  Guidelines  have  been  an  evolving  docu- 
ment. As  more  has  been  learned  about  rDNA 
and  molecular  genetics,  containment  levels 
have  been  significantly  lowered.  Also,  the  de- 
gree of  Federal  ox'ersight  has  been  substantially 
lessened.  Under  the  November  1980  Guidelines, 
virtually  all  responsibility  for  monitoring  com- 
pliance is  placed  on  the  IBCs.  NIH’s  role  will  in- 
x'olve  primarily:  1)  continuing  interpretation  of 
the  Guidelines,  2)  certifying  new  host-vector 
systems,  3)  serving  as  a clearinghouse  of  infor- 
mation, 4)  continuing  risk  assessment  experi- 
ments, and  5)  coordinating  Federal  and  local  ac- 
tivities. 

The  most  significant  short-term  limitation  of 
the  Guidelines  is  the  way  they  deal  with  com- 
mercial applications  and  products  of  rDNA  tech- 
niques. Although  large-scale  containment  levels 
and  related  administrative  procedures  exist, 
there  are  several  reasons  for  questioning  the  ef- 
fectiveness of  the  voluntary  compliance  con- 
cept. The  most  serious  problem  has  been  the 
lack  of  expertise  in  fermentation  technology  on 
RAC.  In  addition,  since  the  Guidelines  are  not 
legally  binding  upon  industry,  the  NIH  lacks  en- 
forcement authority,  although  there  has  been 
no  evidence  of  industrial  noncompliance.  Final- 
ly, because  of  its  role  as  a promoter  of  bio- 


224  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animais 


medical  research,  NIH  cannot  be  expected  to  act 
aggressively  to  fill  this  regulatory  void. 

As  a model  for  societal  decisionmaking  on 
technological  risks,  the  system  created  by  the 
Guidelines  could  serve  as  a valuable  precedent. 
It  does  a reasonable  job  of  combining  substan- 
tive scientific  evaluation  of  technical  issues  with 


procedural  safeguards  designed  to  accommo- 
date social  values  and  to  limit  conflicts  of  in- 
terest. The  only  major  criticism  is  that  proce- 
dural safeguards  and  public  input  were  not  sig- 
nificant factors  when  the  rDNA  problem  was 
first  addressed. 


Other  means  of  regulation 


There  are  three  other  means  available  for 
regulating  molecular  genetic  techniques  and 
their  products— current  Federal  statutes,  tort 
law  and  workmen’s  compensation,  and  State 
and  local  laws.  These  all  may  be  used  to  remedy 
some  of  the  limitations  of  the  Guidelines. 

Federal  statutes 

The  question  of  whether  existing  Federal 
statutes  provide  adequate  regulatory  authority 
first  arose  with  respect  to  rDNA  research.  In 
March  1977,  the  Interagency  Committee  con- 
cluded that  while  a number  of  statutes*  could 
provide  authority  to  regulate  specific  phases  of 
work  with  rDNA,  no  single  one  or  combination 
would  clearly  reach  all  rDNA  research  to  the  ex- 
tent deemed  necessary  by  the  Committee.  Fur- 
thermore, while  some  could  be  broadly  inter- 
preted, the  Committee  believed  that  regulatory 
action  taken  on  the  basis  of  those  interpreta- 
tions would  be  subject  to  legal  challenge. This 
was  the  basis  for  their  conclusion  that  specific 
legislation  was  needed  and  was  one  of  the  rea- 
sons behind  the  legislative  effort  discussed  in 
app.  III-A. 

With  respect  to  commercial  uses  and  prod- 
ucts of  rDNA  and  other  genetic  techniques,  a 
much  more  certain  basis  for  regulation  exists. 
Many  of  the  Federal  environmental,  product 
safety,  and  public  health  laws  are  directed 
toward  industrial  processes  and  products.  To  a 

‘The  Committee  concentrated  on  the  tollowing  statutes:  1)  the 
Occupational  Safety  and  Health  Act  (29  U.S.C.  §651  et.  seq  );  2)  the 
Toxic  Substances  Control  Act  (15  U.S.C.  §2601  et.  seq  );  3)  the  Haz- 
ardous Materials  Transportation  Act  (49  U.S.C.  §1801  et.  seq  );  and 
4)  sec.  361  of  the  Public  Health  Service  Act  (42  U.S.C.  §264). 

^^Interim  Report  of  the  Federal  Interagency  Committee  on  Recom- 
binant DNA  Research.  Suggested  Elements  for  Legislation,  op.  cit. 


large  extent,  the  genetic  technologies  will  pro- 
duce chemicals,  foods,  and  drugs— as  well  as 
pollutant  byproducts— that  will  clearly  come 
within  the  scope  of  these  laws. 'However,  there 
may  be  limitations  in  these  laws  and  questions 
of  their  interpretation  that  may  arise  with  re- 
spect to  the  manufacturing  process,  which 
employs  large  quantities  of  organisms,  and 
when  there  is  an  intentional  release  of  micro- 
organisms into  the  environment— e.g.,  for  clean- 
ing up  pollution.  For  a list  of  pertinent  laws,  see 
table  36.) 

The  Federal  Food,  Drug,  and  Cosmetic  Act 
(FFDCA)  and  section  351  of  the  Public  Health 
Service  Act  (42  U.S.C.  262)  give  FDA  authority 
over  foods,  drugs,  biological  products  (such  as 
vaccines),  medical  devices,  and  veterinary  medi- 
cines. This  authority  will  also  apply  to  those 
products  when  they  are  made  by  genetic  engi- 

Table  36.— Statutes  That  Will  Be  Most  Applicable 
to  Commercial  Genetic  Engineering 


1.  Federal  Food,  Drug,  and  Cosmetic  Act  (21  U.S.C.  §301 
et.  seq.) 

2.  Occupational  Safety  and  Health  Act  (29  U.S.C.  §651  et. 
seq.) 

3.  Toxic  Substances  Control  Act  (15  U.S.C.  §2601  et. 
seq.) 

4.  Marine  Protection,  Research,  and  Sanctuaries  Act  (33 
U.S.C.  §1401  et.  seq.) 

5.  Federal  Water  Pollution  Control  Act,  as  amended  by 
the  Clean  Water  Act  of  1977  (33  U.S.C.  §1251  et.  seq  ) 

6.  The  Clean  Air  Act  (42  U.S.C.  §7401  et.  seq.) 

7.  Hazardous  Materials  Transportation  Act  (49  U.S.C, 
§1801  et.  seq.) 

8.  Solid  Waste  Disposal  Act,  as  amended  by  the 
Resource  Conservation  and  Recovery  Act  of  1976  (42 
U.S.C.  §6901  et.  seq.) 

9.  Public  Health  Service  Act  (42  U.S.C.  §201  et.  seq  ) 

10.  Federal  Insecticide,  Fungicide,  and  Rodenticide  Act  i7 
U.S.C.  §136  et.  seq.) 


SOURCE:  Office  of  Technology  Assessment 


Ch.  11— Regulation  of  Genetic  Engineering  • 225 


neering  methods.  However,  interpreti\e  ques- 
tions ai'ising  out  of  the  unique  nature  of  the 
technologies— such  as  the  type  of  data  nec- 
essary to  show  the  safety  and  efficacy  of  a new 
drug  produced  by  rDNA  techniques— will  have 
to  be  resoKed  by  the  administrative  process  on 
a case-by-case  basis. 

FU.A  has  not  published  any  statements  of  of- 
ficial policy  toward  products  made  by  genetic 
engineering.  Since  it  has  different  statutory  au- 
thority for  different  types  of  products,  it  is  like- 
ly that  regulation  u ill  be  on  a product-by-prod- 
uct basis  through  the  appropriate  FDA  bureau. 
Substances  produced  by  genetic  engineering 
will  generally  be  treated  as  analogous  products 
produced  by  conventional  techniques  with  re- 
spect to  standards  for  chemistry,  pharmacolo- 
g\',  and  clinical  protocols;  howe\  er,  quality  con- 
trols may  have  to  be  modified  to  assure  continu- 
ous control  of  product  purity  and  identity.  In 
addition,  for  the  time  being,  the  Bureau  of 
Drugs  and  the  Bureau  of  Biologies  will  require  a 
new  IMotice  of  Claimed  Investigational  Exemp- 
tion for  a New  Drug  and  a new  New  Drug  Appli- 
cation for  products  made  by  rDN,A  technology, 
e\en  if  identity  with  the  natural  substance  or 
with  a previously  appro\  ed  drug  is  shown.  This 
policy  is  based  on  the  position  that  drugs  or 
biologies  made  by  rDN,A  techniques  have  not 
become  generally  recognized  by  experts  as  safe 
and  effective  and  therefore  meet  the  statutory 
definition  of  a "new  drug. 

FFDCA  also  permits  regulation  of  drug,  food, 
and  device  manufacturing.  Certain  FDA  regula- 
tions, called  Good  Manufacturing  Practices,  are 
designed  to  assure  the  quality  of  these  products. 
FDA  may  have  to  revise  these  to  accommodate 
genetic  technologies;  it  has  the  authority  to  do 
so.  It  probably  does  not  have  the  authority  to 
use  these  regulations  to  address  any  risks  to 
workers,  the  public,  or  the  enx  ironment,  since 
FFDCA  is  designed  to  protect  the  consumer  of 
the  regulated  product. 


“Minutes  of  the  Industrial  Practices  Subcommittee  of  the  Fed- 
eral Interagency  Advisory  Committee  on  Recombinant  DNA  Re- 
search, Dec.  16,  1980,  p.  3. 

•Sec.  201(p)  of  the  FFDCA  (21  U.S.C.  §321(p))  defines  a new  drug 
as  "anv  drug  . . . the  composition  of  which  is  such  that  such  drug 
is  not  generally  recognized,  among  experts  qualified  by  scientific 
training  and  experience  ...  as  safe  and  effective 


The  statute  most  applicable  to  worker  health 
and  safety  is  the  Occupational  Safety  and  Health 
Act,  which  grants  the  Secretary  of  Labor  broad 
power  to  reciuire  employers  to  provide  a safe 
workplace  for  their  employees.  This  power  in- 
cludes the  ability  to  require  an  employer  to 
modify  work  practices  and  to  install  control 
technology'.  The  statute  creates  a general  duty 
on  employers  to  furnish  their  employees  with  a 
workplace  "free  from  recognized  hazards  that 
are  causing  or  are  likely  to  cause  death  or  seri- 
ous physical  harm,”  and  it  requires  employers 
to  comply  with  occupational  safety  and  health 
standards  set  by  the  Secretary  of  Labor.  Accord- 
ing to  a recent  Supreme  Court  case,  a standard 
may  be  promulgated  only  on  a determination 
that  it  is  "reasonably  necessary  and  appropriate 
to  remedy  a significant  risk  of  material  health 
impairment.”^'*  Because  these  fairly  stringent  re- 
quirements limit  the  Act’s  applicability  to 
recognized  hazards  or  significant  risks,  the 
statute  could  not  be  used  to  control  manufactur- 
ing where  the  genetic  techniques  presented  on- 
ly hypothetical  risks.  However,  it  should  be  ap- 
plicable to  large-scale  processes  using  known 
human  toxins,  pathogens,  or  their  DNA. 

The  Secretary  of  Labor  is  also  directed  to  ac- 
count for  the  "urgency  of  the  need”  in  es- 
tablishing regulatory  priorities.  How  the  De- 
partment of  Labor  will  view  genetic  technol- 
ogies within  its  scale  of  priorities  remains  to  be 
seen.  NIOSH,  the  research  organization  created 
by  this  statute,  has  been  studying  rDNA  produc- 
tion methods  to  determine  what  risks,  if  any, 
are  being  faced  by  workers.  It  has  conducted 
fact-finding  inspections  of  several  manufac- 
turers, and  it  is  planning  a joint  project  with 
EPA  to  assess  the  adequacy  of  current  control 
technology.  In  addition,  a group  established  by 
the  Center  for  Disease  Control  (CDC)  together 
with  NIOSH  will  be  making  recommendations 
on:  1)  the  medical  surveillance  of  potentially  ex- 
posed workers,  2)  the  central  collection  and 
analysis  of  medical  data  for  epidemiological  pur- 
poses, and  3)  the  establishment  of  an  emergency 
response  team.®^ 

^“Industrial  Union  Department,  AFL-CIO  v.  American  Petroleum 
Institute,  100  S.Ct.  2844,2863,  1980. 

’^Minutes  of  the  Industrial  Practices  Subcomittee  of  the  Federal 
Interagency  Advisory  Committee  on  Recombinant  DNA  Research, 
Dec.  16,  1980,  op.  cit.,  p.  6. 


226  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


The  Toxic  Substances  Control  Act  (TSCA)  was 
intended  by  Congress  to  fill  in  the  gaps  in  the 
other  environmental  laws.  It  authorizes  EPA  to 
acquire  information  on  “chemical  substances”  in 
order  to  identify  and  evaluate  potential  hazards 
and  then  to  regulate  the  production,  use,  distri- 
bution, and  disposal  of  those  substances. 

A “chemical  substance”  is  defined  under  sec- 
tion 3(2)  of  this  Act  as  “any  organic  or  inorganic 
substance  of  a particular  molecular  identity,”  in- 
cluding "any  combination  of  such  substances  oc- 
curring in  whole  or  in  part  as  a result  of  a chem- 
ical reaction  or  occurring  in  nature.”* *  This 
would  include  DNA  molecules;  however,  it  is 
unclear  if  the  definition  would  encompass  gene- 
tically engineered  organisms.  In  promulgating 
its  Inventory  Reporting  Regulations  under 
TSCA  on  December  23,  1977,  EPA  took  the  fol- 
lowing position  in  response  to  a comment  that 
commercial  biological  preparations  such  as 
yeasts,  bacteria,  and  fungi  should  not  be  con- 
sidered chemical  substances;^® 

The  Administrator  disagrees  with  this  com- 
ment ....  This  definition  [of  chemical  sub- 
stance] does  not  exclude  life  forms  which  may 
be  manufactured  for  commercial  purposes  and 
nothing  in  the  legislative  history  would  suggest 
otherwise. 

However,  in  a December  9,  1977,  letter  re- 
sponding to  a Senate  inquiry,  EPA  Administra- 
tor Douglas  M.  Costle  stated:®^ 

[Allthough  there  is  a general  consensus  that  re- 
combinant DNA  molecules  are  “chemical  sub- 
stances” within  the  meaning  of  section  3 of 
TSCA,  it  is  not  at  all  clear  whether  a host  or- 
ganism containing  recombined  DNA  molecules 
fits— or  was  intended  to  fit— that  definition  .... 

If  such  organisms  are  subject  to  TSCA  on  the 
grounds  that  they  are  a “combination  of  ... 
substances  occurring  in  whole  or  in  part  as  a 
result  of  a chemical  reaction,”  the  Agency  might 
logically  have  to  include  all  living  things  in  the 
definition  of  "chemical  substance”— an  inter- 

‘Substances  subject  solely  to  FFDCA  or  tbe  Federal  Insecticide, 
Fungicide,  and  Rodenticide  Act  are  excluded  from  this  definition. 
“42  F.R.,  64572,  64584,  Dec.  23,  1977. 

^'Letter  to  Adlai  E.  Stevenson,  Chairman,  Subcommittee  on  Sci- 
ence, Technology,  and  Space,  U.S.  Senate  Committee  on  Com- 
merce, Science,  and  Transportation,  in  Oversight  Report,  Recombi- 
nant DNA  Research  and  Its  Applications,  95th  Cong.,  2d  sess.,  Au- 
gust 1978,  p.88. 


pretation  which  I am  confident  the  Congress 

neither  contemplated  nor  intended. 

If  EPA  were  to  take  the  broader  interpreta- 
tion, and  if  that  were  to  survive  any  legal  chal- 
lenge, TSCA  would  have  great  potential  for  reg- 
ulating commercial  genetic  engineering  by  reg- 
ulating the  organisms.  Under  section  4 of  this 
Act,  EPA  can  adopt  rules  requiring  the  testing  of 
chemical  substances  that  “may  present  an  un- 
reasonable risk”*  to  health  or  the  environment 
when  existing  data  are  insufficient  to  make  a 
determination.  Under  section  5,  the  manu- 
facturer of  a new  chemical  substance  is  re- 
quired to  notify  EPA  90  days  before  beginning 
production  and  to  submit  any  test  data  available 
on  the  chemical’s  health  or  environmental  ef- 
fects. If  EPA  decides  that  the  data  are  insuffi- 
cient for  evaluating  the  chemical’s  effects  and 
that  it  "may  present  an  unreasonable  risk”  or 
will  be  produced  in  substantial  quantities,  the 
chemical  substance’s  manufacture  or  use  can  he 
restricted  or  prohibited.  Under  section  0,  EP.A 
can  prohibit  or  regulate  the  manufacture  or  use 
of  any  chemical  substance  that  "presents,  oi'  will 
present  an  unreasonable  risk  of  injury  to  health 
or  the  environment.” 

As  with  the  Occupational  Safety  and  Health 
Act,  the  scientific  evidence  probably  does  not 
support  a finding  that  most  genetically  en- 
gineered molecules  or  organisms  present  an  un- 
reasonable risk.  On  the  other  hand,  the  stand- 
ard in  section  5— may  present  an  unreasonable 
risk— and  the  requirement  for  a premanulae- 
turing  notice  would  permit  El’A  to  e\aluat(* 
cases  where  genetically  engineered  mici’o-orga- 
nisms  were  proposed  to  he  released  into  the 
environment. 

Several  other  environmental  statutes  w ill  ap- 
ply, mainly  with  I'espect  to  pollutants,  wastes, 
or  hazardous  materials.**  The  Marine'  I’rotee- 


‘ The  term  'unrea.sonahle  I'i.sk  " is  not  delineil  in  the  sl.ilwlr 
However,  the  legislative  histoi-y  indicates  that  its  drtcrmin.itinn  in 
volves  balancing  the  probability  that  harm  will  occur  and  the 
magnitude  and  severity  ot  that  harm,  against  the  cltci  t ol  the  pro 
posed  regulatory  action  and  the  a\ailahilit\  to  socii4\  ol  thi'  bene 
fits  of  the  substance. “ 

'“H.  Kept.  94-1341. 94lh  ( ong  . 2d  sess  1976  pp  l.l  t,'. 

* ‘ Two  consumer  protection  statutes  w ere  considei  I'd  lint  w i-i  e 
determined  to  he  x irlualiv  inapplicable  I hese  v\eie  the  lc•del.ll 

Hazardous  Substances  ,\ct  ( 15  t ' S I §t2litet  sei|  I anil  the  ( on 
sumer  I’roduct  Safety  Act  (15  tl.S  ( §2tl5l  el  sei|  I 


Ch.  11 — Regulation  of  Genetic  Engineering  • 227 


tion,  Research,  and  Sanctuaries  Act  prohibits 
ocean  dumping  without  an  EPA  permit  of  any 
material  that  would  “unreasonably  degrade  or 
endanger  human  health,  welfare,  or  amenities, 
or  the  mai’ine  en\  ironment,  ecological  systems, 
or  economic  potentialities.”^®  "Material"  is  de- 
fined as  "matter  of  any  kind  oi'  description,  in- 
cluding . . . biological  and  laboratory  waste 
. . . and  industrial  . . . and  other  waste."'"’  The 
Federal  Water  Pollution  Control  .Act  regulates 
the  discharge  of  pollutants  (which  include  bio- 
logical materials)  into  LfS.  waters,  and  the  Solid 
Waste  Disposal  ,Act  regulates  hazardous  wastes. 
The  Clean  .Air  .Act  regulates  the  discharge  of  air 
pollutants,  which  includes  biological  materials. 
Especially  applicable  is  section  112  (42  U.S.C^  § 
7412),  w hich  allows  EP.A  to  set  emission  stand- 
ards for  hazardous  air  pollutants— those  for 
which  standards  have  not  been  set  under  other 
sections  of  the  Act  and  which  "may  reasonably 
be  anticipated  to  result  in  an  increase  in  mortali- 
ty or  an  increase  in  serious  irre\  ersible,  or  in- 
capacitating re\ersible,  illness.”  The  Hazardous 
Materials  Transportation  .Act  co\ers  the  inter- 
state transportation  of  dangerous  articles,  in- 
cluding etiologic  (disease-causing)  agents.  The 
Secretary  of  Transportation  may  designate  as 
hazardous  any  material  that  he  finds  "may  pose 
an  unreasonable  risk  to  health  and  safety  or 
property”  when  transported  in  commerce  in  a 
particular  quantity  and  form.^’ 

Section  361  of  the  Public  Health  Ser\  ice  Act 
(42  U.S.C.  §264)  authorizes  the  Secretary  of 
HEW  (now  DHHS)  to  “.  . . make  and  enforce 
such  regulations  as  in  his  judgment  are  neces- 
sary to  pre\  ent  the  introduction,  transmission, 
or  spread  of  communicable  diseases  . . . .”  Be- 
cause of  the  broad  discretion  given  to  tbe  Sec- 
retary, it  bas  been  argued  that  this  section  pro- 
\'ides  sufficient  authority  to  control  all  rDNA  ac- 
tivities. * Others  ha\  e argued  that  its  purpose  is 
to  protect  only  human  health;  for  regulations  to 
be  \ alid,  there  would  have  to  be  a supportable 
finding  of  a connection  between  rDNA  and 


”33  U.S.C.  § 1412. 

“33  U.S.C.  § 1402(c). 

“49  U.S.C.  § 1803. 

'On  Nov.  11,  1976,  the  Natural  Resources  Defense  Council  and 
the  Environmental  Defense  Fund  petitioned  the  Secretary  of  HEW 
to  promulgate  regulations  concerning  rDNA  under  this  Act. 


human  disease.  In  any  event,  HEW  declined  to 
promulgate  any  regulations. 

The  following  conclusions  can  therefore  be 
made  on  the  applicability  of  existing  statutes. 
First,  tbe  products  of  genetic  technologies— 
such  as  drugs,  chemicals,  pesticides,**  and 
foods— u'ould  clearly  be  covered  by  statutes 
already  covering  these  generic  categories  of 
materials.  Second,  uncertainty  exists  for  regu- 
lating either  production  methods  using  en- 
gineered micro-organisms  or  their  intentional 
release  into  the  environment,  when  risk  has  not 
been  clearly  demonstrated.  Third,  the  regu- 
latory agencies  have  begun  to  study  the  situa- 
tion but  have  not  promulgated  specific  regu- 
lations. Fourth,  since  regulation  will  be  dis- 
persed throughout  several  agencies,  there  may 
be  conflicting  interpretations  unless  active  ef- 
forts are  made  by  the  Federal  Interagency  Com- 
mittee to  develop  a comprehensive,  coordinated 
approach. 

Tort  law  and  workmen's  compensation 

Statutes  and  regulations  are  usually  directed 
at  preventing  certain  types  of  conduct.  While 
tort  law  strives  for  the  same  goal,  its  primary 
purpose  is  to  compensate  injuries.  (A  tort  is  a 
civil  wrong,  other  than  breach  of  contract,  for 
which  a court  awards  damages  or  other  relief.) 
By  its  nature,  tort  law  is  quite  flexible,  since  it 
has  been  dev'eloped  primarily  by  the  courts  on  a 
case-by-case  basis.  Its  basic  principles  can  easily 
be  applied  to  cases  where  injuries  have  been 
caused  by  a genetically  engineered  organism, 
product,  or  process.  It  therefore  can  be  applied 
to  cases  involving  genetic  technologies  as  a 
means  of  compensating  injuries  and  as  an  incen- 
tive for  safety-conscious  conduct.  The  most  ap- 
plicable concepts  of  tort  law  are  negligence  and 
strict  liability.  (A  related  body  of  law— work- 
men’s compensation— is  also  pertinent.) 

Negligence  is  defined  as  conduct  (an  act  or  an 
omission)  that  involves  an  unreasonable  risk  of 
harm  to  another  person.  For  the  injured  party 
to  be  compensated,  he  must  prove  in  court  that: 
1)  the  defendant’s  conduct  was  negligent,  2)  the 


••Pesticides  are  subject  to  the  Federal  Insecticide,  Futigicide, 
and  Rodenticide  Act,  7 U.S.C.  § 136  et.  seq.. 


228  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


defendant’s  actions  in  fact  caused  the  injury, 
and  3)  the  injury  was  not  one  for  which  com- 
pensation should  he  denied  or  limited  because 
of  overriding  policy  reasons. 

Because  of  the  newness  of  genetic  technol- 
ogy, legal  standards  of  conduct  (e.g.,  what  con- 
stitutes unreasonable  risk)  have  not  been  ar- 
ticulated by  the  courts.  If  a case  were  to  arise,  a 
court  would  undoubtedly  look  first  to  the 
Guidelines.  Even  if  a technique  other  than  rDNA 
were  involved,  they  would  provide  a general 
conceptual  framework  for  good  laboratory  and 
industrial  techniques.  Other  sources  for  stand- 
ards of  conduct  include:  1)  CDC’s  guidelines  for 
working  with  hazardous  agents;  2)  specific  Fed- 
eral laws  or  regulations,  such  as  those  under  the 
Public  Health  Service  Act  covering  the  inter- 
state transportation  of  biologic  products  and 
etiologic  agents;  and  3)  industrial  or  profes- 
sional codes  or  customary  practices,  such  as 
generally  accepted  containment  practices  in  the 
pharmaceutical  industry  or  in  a microbiology 
laboratory.  Compliance  with  these  standards, 
however,  does  not  foreclose  a finding  of  neg- 
ligence, since  the  courts  make  the  ultimate  judg- 
ment of  what  constitutes  proper  conduct.  In 
several  cases,  courts  have  decided  that  an  entire 
industry  or  profession  has  lagged  behind  the 
level  of  safe  practices  demanded  by  society.* 
Conversely,  noncompliance  with  existing  stand- 
ards almost  surely  will  result  in  a finding  of 
negligence,  if  the  other  elements  are  also  pres- 
ent. 

Causation  may  be  difficult  to  prove  in  a case 
involving  a genetically  engineered  product  or 
organism.  In  the  case  of  injury  caused  by  a path- 
ogenic micro-organism— e.g.,  it  may  be  difficult 
to  isolate  and  identify  the  micro-organism  and 
virtually  impossible  to  trace  its  origin,  especially 
if  it  had  only  established  a transitory  ecological 
niche.  In  addition,  it  might  be  difficult  to 
reconstruct  the  original  situation  to  determine 
if  the  micro-organism  simply  escaped  despite 


•For  example,  see:  The  T.  J.  Hooper,  60  F.  2d  737  (2d  Cir.  1932), 
concerning  tugboats;  and  Helling  v.  Carey,  519  P.  2d  981  (1974), 
where  the  court  held  that  the  general  practice  among  ophthalmo- 
logists of  not  performing  glaucoma  tests  on  asymptomatic  patients 
under  40  (because  they  had  only  a one  in  25,000  chance  of  having 
the  disease)  would  not  prevent  a finding  of  negligence  when  such 
a patient  developed  the  disease. 


precautions  or  if  culpable  human  action  was  in- 
volved. On  the  other  hand,  if  a micro-organism 
or  toxin  is  identified,  it  may  be  so  unique 
because  of  its  engineering  that  it  can  he  readily 
associated  with  a company  known  to  produce  it 
or  with  a scientist  known  to  be  working  with 
it.** 

The  law  recognizes  that  not  every  negligent 
act  or  omission  that  causes  harm  should  result 
in  liability  and  compensation— e.g.,  the  concept 
of  "foreseeable”  harm  serves  to  limit  a de- 
fendant’s liability.  The  underlying  social  policy 
is  that  the  defendant  should  not  he  liable  for  in- 
juries so  random  or  unlikely  as  to  he  not  rea- 
sonably foreseeable.  This  determination  is  made 
by  the  court.  In  the  case  of  a genetically  en- 
gineered organism,  extensive  harm  would  prob- 
ably be  foreseeable  because  of  the  organism’s 
ability  to  reproduce;  how  that  harm  could  occur 
might  not  be  foreseeable. 

Unlike  negligence,  strict  liability  does  not  re- 
quire a finding  that  the  defendant  breached 
some  duty  of  care  owed  to  the  injured  person; 
the  fact  that  the  injury  was  caused  by  the  de- 
fendant’s conduct  is  enough  to  impose  liability 
regardless  of  how  carefully  the  activity  was 
done.  For  this  doctrine  to  apply,  the  activity 
must  be  characterized  as  "abnormally  dan- 
gerous.” To  determine  this,  a court  would  look 
at  the  following  six  factors,  no  one  of  which  is 
determinative:"*^ 

1.  existence  of  a high  risk  of  harm, 

2.  great  gravity  of  the  hai  in  if  it  occurs, 

3.  inability  to  eliminate  the  risk  by  exei’cising 
reasonable  care. 


•’ll' several  companies  were  working  with  Ihe  micro-organiMn 
it  could  be  impossible  to  pro\-e  which  company  prodm  ed  Ihe  par 
ticular  ones  that  caused  the  harm.  V recent  ( alilornia  Supreme 
Court  case,  Sindell  v.  Ahholl  l.ahoratories,  26  ( al  3d  5HK  l9Mtl 
could  pro\'ide  a way  around  this  |)roblem  it  the  new  lheor\  ol 
liahility  that  it  establishes  hecomes  widely  accepted  Iw  coiii  ls  m 
other  jurisdictions.  Ihe  Court  ruled  Ih.it  women  whose  mothiTs 
had  taken  diethylstilbestrol.  a drug  that  allegedly  c.iiised  c.iiii  ei  m 
their  daughters,  could  proceed  to  trial  .igainst  m.iiuit.iclurers  ol 
the  drug,  even  though  most  ol  Ihe  plaintitls  would  not  be  .ihle  to 
show  which  |)arlicular  manutaclurers  produced  Ihe  drug  I he 
Court  said  that  wh(>n  the  delendani  manut.iclurers  h.id  ,i  suhslaiv 
tial  share  of  Ihe  product  market,  liahililv  it  lound  would  he  ap 
portioned  among  the  defendants  on  Ihe  h.isis  ol  their  m.irkel 
share,  A particular  defendant  could  esc.ipe  li.ihilil\  oiiK  h\ 
proving  it  could  not  ha\c  made  Ihe  drug 
^^Restatemenl  (Second)  of  Torts  §5211 1 19761 


Ch.  11 — Regulation  of  Genetic  Engineering  • 229 


4.  extent  to  w hich  the  acti\  ity  is  not  common, 

5.  inappro[)i'iateness  of  the  acti\  itv  to  the 
place  where  it  is  done,  and 

6.  the  acti\  itv’s  value  to  the  community. 

C'ii\en  the  current  consensus  about  the  risks 
of  genetic  technicjues,  it  would  he  difficult  to 
argue  that  the  doctrine  of  strict  liability  should 
apply.  How  e\er,  in  the  extremely  unlikely  e\ent 
that  a serious,  w idespread  injury  does  occur', 
that  alone  would  probably  suppoi't  a court's  de- 
termination that  the  activity  was  abnormally 
dangerous,  I'egai'dless  of  its  pi'ohahility.  In  such 
cases,  the  courts  have  generally  relied  on  the 
principle  of  “enterprise  liability"— that  those  en- 
gaged in  an  enterprise  should  hear  its  costs,  in- 
cluding the  costs  of  injuries  to  others. 

For  either  negligence  or  strict  liability,  the 
person  causing  the  harm  is  liable.  L'nder  the 
legal  principle  of  respondeat  superior,  liability  is 
also  imputed  fi'om  the  original  actor  to  people 
or  entities  w ho  have  a special  relationship  with 
him— e.g.,  employers.  I'hus,  a corporation  can 
he  liable  for  the  torts  of  its  scientists  or  produc- 
tion workers.  Similarly,  a university,  an  IBC,  a 
Biological  Safetv  Officer,  and  a PI  would  prob- 
ably he  liable  for  the  torts  of  scientists  and  stu- 
dents under  their  direction. 

.Another  body  of  law  designed  to  compensate 
injuries  deserves  brief  mention.  Workmen’s 
compensation  is  a statutory  scheme  adopted  by 
the  States  and— for  specific  occupations  or  cir- 
cumstances—by  the  Federal  Government  to 
compensate  injuries  without  a need  for  showing 
fault.  The  employee  need  only  show  that  the  in- 
jury was  job-related.  He  is  then  compensated  by 
the  employer  or  the  employer's  insurance  com- 
pany. It  would  clearly  apply  to  genetic  engineer- 
ing. 

Tort  law  and  workmen’s  compensation  will 
be  available  to  compensate  any  injuries  re- 
sulting from  the  use  of  molecular  genetic  tech- 
niques, especially  from  their  commercial  appli- 
cation. Tort  law  may  also  indirectly  prevent 
potentially  hazardous  actions,  although  the  de- 


■•’R.  Dworkin.  "Biocatastrophe  and  the  Law:  Legal  .Aspects  of  Re- 
combinant DN.A  Research,"  in  The  Recombinant  D\'A  Debate, 
Jackson  and  Stitch  (eds.)  (Englewood  Cliffs,  X.J.:  Prentice-Hall,  Inc. 
1979),  pp.  219,  223. 


terrent  effect  of  compensation  is  less  efficient 
than  direct  regulation— e.g.,  the  threat  of  law- 
suits will  not  necessarily  discourage  high-risk 
activities  where  problems  of  proof  make  re- 
covery unlikely,  where  the  harm  may  be  small 
and  widespread  (as  with  mild  illness  suffered  by 
a large  number  of  people),  or  where  profits  are 
less  than  the  cost  of  prevention  but  greater  than 
expected  damage  awards  and  legal  costs. 

Tort  law  has  two  other  limitations.  First,  tort 
litigation  involves  high  costs  to  the  plaintiff,  and 
indirectly  to  society.  Second,  it  cannot  adequate- 
ly compensate  the  victims  of  a catastrophic  sit- 
uation where  liability  would  bankrupt  the 
defendant. 

State  and  local  law 

L’nder  the  10th  amendment  to  the  Constitu- 
tion, all  powers  not  delegated  to  the  Federal 
Government  are  reserved  for  the  States  or  the 
people.  One  of  those  is  the  power  of  the  States 
and  municipalities  to  protect  the  health,  safety, 
and  welfare  of  their  citizens.  Thus,  they  can 
regulate  genetic  engineering. 

The  reasons  espoused  in  favor  of  local  regula- 
tion are  based  on  the  traditional  concept  of  local 
autonomy;  those  most  likely  to  suffer  any 
adv’erse  affects  of  genetic  engineering  should 
control  it.  Also,  local  and  State  governments  are 
usually  more  accessible  to  public  input  than  the 
Federal  Government.  Consequently,  judgments 
on  the  acceptability  of  the  risks  will  more 
precisely  reflect  the  will  of  the  segment  of  the 
public  most  directly  affected. 

A number  of  arguments  have  been  made 
against  local  as  opposed  to  Federal  regulation. 
The  primary  one  is  that  regulation  by  States  and 
communities  would  give  rise  to  a random  patch- 
work  of  confusing  and  conflicting  controls.  In 
addition.  States  and  especially  localities  may  not 
have  the  same  access  as  the  Federal  Govern- 
ment to  the  expertise  that  should  be  used  in  the 
formulation  of  rational  controls.  Finally,  any 
risks  associated  with  rDNA  or  other  techniques 
are  not  limited  by  geographic  boundaries; 
therefore,  they  ought  to  be  dealt  with  national- 
ly. The  above  arguments  reflect  the  position 
that  regulation  of  genetic  technologies  is  a na- 


230  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


tional  issue  that  can  be  handled  most  effectively 
at  the  Federal  level. 

A few  jurisdictions  have  used  their  authority 
in  the  case  of  rDNA.  * The  most  comprehensive 
regulation  was  created  by  the  States  of  Mary- 

'Cambridge,  Mass.,  established  a citizens'  study  group  that  rec- 
ommended that  researchers  be  subject  to  some  additional  re- 
strictions beyond  those  of  the  Guidelines.  These  were  embodied  in 
an  ordinance  passed  by  the  City  Council  on  Feb.  7,  1977.  Berkeley, 
Calif.,  passed  an  ordinance  requiring  private  research  to  conform 
to  the  Guidelines.  Similar  ordinances  or  resolutions  were  passed 
by  Princeton,  N.J.,  Amherst,  Mass.,  and  Emeryville,  Calif. 


Conclusion  

The  initial  question  with  respect  to  regulating 
genetic  engineering  is  how  to  define  the  scope 
of  the  problem.  This  will  depend  largely  on 
what  groups  are  involved  in  that  process  and 
how  they  view  the  nature,  magnitude,  and 
acceptability  of  the  risks.  Similarly,  the  means 
of  addressing  the  problem  will  be  determined 
by  how  it  is  defined  and  who  is  involved  in  the 
actual  decisionmaking  process.  For  these  rea- 
sons, it  is  important  that  regulatory  mechanisms 
combine  scientific  expertise  with  procedures  to 
accommodate  the  values  of  those  bearing  the 
risk  so  that  society  may  have  confidence  in 
those  mechanisms. 

Currently,  genetic  techniques  and  their  prod- 
ucts are  regulated  by  a combination  of  the 


Issue  ai 

ISSUE:  How  could  Congress  address  the 

risks  associated  with  genetic  en- 
gineering? 

A number  of  options  are  available,  ranging 
from  deregulation  through  comprehensive  new 
regulation.  An  underlying  issue  for  most  of 
these  options  is:  What  are  the  constitutional 
constraints  placed  on  congressional  regulation 
of  molecular  genetic  techniques,  particularly 
when  they  are  used  in  research?  (This  is  dis- 
cussed in  app.  III-B.) 


land  and  New  York.'*^  Currently,  there  is  little, 
if  any,  effort  on  the  State  or  local  level  to  pass 
laws  or  ordinances  covering  rDNA  or  similar 
genetic  techniques,  and  there  is  little  activity 
under  the  existing  laws. 


^“Annotated  Code  o f Maryland,  ;irt.  43  §§  898-910  (supp.  19781 
“^McKinnev’s  Consolidated  Laws  of  New  )'ork,  Public  MimIiIi  Law  . 
art.  32-A  §§3220-3223  (supp.  1980) 


Guidelines,  Federal  statutes  protecting  health 
and  the  environment,  some  State  or  local  laws, 
and  the  judicially  created  law  of  torts,  which  is 
available  to  compensate  injuries  after  they  oc- 
cur. In  most  cases,  this  system  appears  adequate 
to  deal  with  the  risks  to  health  and  the  en\  iron- 
ment.  However,  there  is  some  concern  regard- 
ing commercial  applications  for  the  following 
reasons:  1)  the  voluntary  applicability  of  the 
Guidelines  to  industry,  2)  RAC's  insufficient  ex- 
pertise in  fermentation  technology,  3)  the  po- 
tential interpretive  problems  in  apjilying  ex- 
isting law  to  the  workplace  and  to  situations 
where  , micro-organisms  are  intentionally  re- 
leased into  the  enviornment,  and  4)  the  absence 
of  a definitive  regulatory  posture  l)v  the 
agencies. 


Options 

OPTIONS: 

A:  Congress  could  maintain  the  status  quo  hy  let- 
ting NIH  and  the  regulatory  agertcies  set  the 
Federal  policy. 

This  option  requires  Congress  to  detcMTiiine 
that  legislation  to  remedy  the  limitations  in  cur- 
rent Federal  oversight  would  result  in  unnec«*s- 
sary  and  burdensome  regulation.  No  knov\ti 
harm  to  health  or  the  em  iionnuMit  has  (m  • 
curred  under  the  current  system,  and  the  agen- 
cies generally  have  significant  legal  authoiit\ 


Ch.  11— Regulation  of  Genetic  Engineering  • 231 


and  expertise  that  should  permit  them  to  adapt 
to  most  new  problems  posed  by  genetic  engi- 
neering. rhe  agencies  ha\e  been  consulting 
with  each  other  through  the  Interagency  Com- 
mittee, and  the  three  agencies  that  will  play  the 
most  important  role  in  regulating  large-scale 
commercial  acti\  ities— FDA,  OSHA,  and  ERA— 
ha\  e been  studying  the  situation. 

The  disad\  antages  of  this  option  are  the  lack 
of  a centralized,  uniform  Federal  response  to 
the  problem,  and  the  possibility  that  risks 
associated  with  commercial  applications  will  not 
be  adequately  addressed.  Certain  applications, 
such  as  the  use  of  micro-organisms  for  oil  re- 
covery are  not  unequix  ocably  regulated  by  cur- 
rent statutes;  broad  interpretations  of  statutory 
language  in  order  to  reach  these  situations  may 
be  overturned  in  court.  Conflicting  or  redun- 
dant regulations  of  different  agencies  would 
result  in  unnecessary  burdens  on  those  regu- 
lated. In  addition,  some  commercial  acti\ity  is 
now  at  the  pilot  plant  stage,  but  the  responsible 
agencies  have  yet  to  establish  official  policy  and 
to  devise  a coordinated  plan  of  action. 

B:  Congress  could  require  that  the  Federal  Inter- 
agency Advisory  Committee  on  Recombinant 
Di\A  Research  prepare  a comprehensive  re- 
port on  its  members'  collective  authority  to 
regulate  rDi\A  and  their  regulatory  intentions. 

The  Industrial  Practices  Subcommittee  of  this 
Committee  has  been  studying  agency  authority 
over  commercial  rDNA  activities.  Presently, 
there  is  little  official  guidance  on  regulatory  re- 
quirements for  companies  that  may  soon  mar- 
ket products  made  by  rDMA  methods.— e.g., 
companies  are  building  fermentation  plants 
without  knowing  what  design  or  other  require- 
ments OSHA  may  mandate  for  worker  safety. 
As  was  stated  by  former  OSHA  head,  Dr.  Eula 
Bingham,  it  will  take  at  least  2 years  for  OSHA  to 
set  standards,  if  the  current  NIOSH  study  shows 
a need  for  them.^® 

A congressionally  mandated  report  would 
assure  full  consideration  of  these  issues  by  the 
agencies  and  expedite  the  process.  It  could  in- 


■•^Letter  from  Dr.  Eula  Bingham.  .Assistant  Secretary  for  Occupa- 
tional Safety  and  Health,  to  Dr.  Donald  Fredrickson.  Director,  NIH, 
Sept.  24,  1980. 


elude  the  following:  1)  a section  prepared  by 
each  agency  that  assesses  its  statutory  authority 
and  articulates  what  activities  and  products  will 
be  considered  to  come  within  its  jurisdiction,  2) 
a summary  section  that  evaluates  the  adequacy 
of  existing  Federal  statutes  and  regulations  as  a 
whole  with  respect  to  commercial  genetic  en- 
gineering, and  3)  a section  proposing  any  specif- 
ic legislation  considered  to  be  necessary. 

Tbe  principal  disadvantages  of  this  option  are 
that  it  may  be  unnecessary  and  impractical.  The 
agencies  are  studying  the  situation,  which  must 
be  done  before  they  can  act.  Also,  it  is  often 
easier  and  more  efficient  to  act  on  each  case  as 
it  arises,  rather  than  on  a hypothetical  basis 
before  tbe  fact. 

C:  Congress  could  require  Federal  monitoring  of 
all  rDNA  activity  for  a limited  number  of 
years. 

This  option  represents  a “wait  and  see”  posi- 
tion by  Congress  and  the  middle  ground  be- 
tween the  status  quo  and  full  regulation.  It  rec- 
ognizes and  balances  the  following  factors:  1) 
the  absence  of  demonstrated  harm  to  human 
health  or  the  environment  from  genetic  en- 
gineering; 2)  the  continuing  concern  that  genet- 
ic engineering  presents  risks;  3)  the  lack  of  suf- 
ficient knowledge  from  which  to  make  a final 
judgment;  4)  the  existence  of  an  oversight  mech- 
anism that  seems  to  be  working  well,  but  that 
has  clear  limitations  with  respect  to  commercial 
activities;  5)  the  virtual  abolition  of  Federal 
monitoring  of  rDNA  activities  by  the  recent 
amendments  to  the  Guidelines;  and  6)  the  ex- 
pected increase  in  commercial  genetic  engineer- 
ing activities. 

Monitoring  involves  the  collection  and  eval- 
uation of  information  about  an  activity  in  order 
to  know  what  is  occurring,  to  determine  the 
need  for  other  action,  and  to  be  able  to  act  if 
necessary.  More  specifically,  this  option  would 
provide  a data  base  that  could  be  used  for:  1)  de- 
termining the  effectiveness  of  voluntary  compli- 
ance with  the  Guidelines  by  industry  and  man- 
datory compliance  by  Federal  grantees,  2)  de- 
termining the  quality  and  consistency  of  IBC  de- 
cisions and  other  actions,  3)  continuing  a formal 
risk  assessment  program,  4)  identifying  vague 


232  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


or  conflicting  provisions  of  the  Guidelines  for 
revision,  5)  identifying  emerging  trends  or  prob- 
lems, and  6)  tracing  any  long-term  adverse  im- 
pacts on  health  or  the  environment  back  to 
their  sources. 

The  obvious  disadvantages  of  this  option  are 
the  increased  paperwork  and  effort  by  scien- 
tists, universities,  corporations,  and  the  Federal 
Government.  Those  working  with  rDNA  would 
have  to  gather  the  required  information  peri- 
odically and  prepare  reports,  which  would  be 
filed  by  the  sponsoring  institution  with  a 
designated  existing  Federal  agency.  A wide- 
range  of  information  would  be  required  for 
each  project.  The  agency  would  have  to  process 
the  reports  and  take  other  actions,  such  as  pre- 
paring an  annual  report  to  Congress,  to  imple- 
ment the  underlying  purposes  of  this  option. 
Additional  manpower  would  most  likely  be 
needed  by  that  agency. 

A statute  implementing  this  option  could  in- 
clude the  following  elements:  1)  periodic  collec- 
tion of  information  in  the  form  of  reports  from 
all  institutions  in  the  United  States  that  sponsor 
any  work  with  rDNA,  2)  active  evaluation  of 
that  information  by  the  collecting  agency,  3)  an- 
nual reports  to  Congress,  and  4)  a sunset  clause. 
Important  information  would  include:  1)  the 
sponsoring  institution's  name;  2)  all  places 
where  it  sponsors  the  research;  and  3)  a tabular 
or  other  summary  that  discloses  for  each  proj- 
ect continuing  or  completed  during  the  report- 
ing period:  the  culture  volume,  the  source  and 
identity  of  the  DNA  and  the  host-vector  system, 
the  containment  levels,  and  other  information 
deemed  necessary  to  effect  the  purposes  of  the 
act.  The  statute  could  also  require  employers  to 
institute  and  report  on  a worker  health  sur- 
veillance program. 

For  this  option  to  work,  the  monitoring  agen- 
cy would  have  to  take  an  active  role  in  eval- 
uating the  data.  It  should  have  the  authority  to 
require  amendments  to  the  reports  when  any 
part  is  vague,  incomplete,  or  inconsistent  with 
another  part.  It  could  also  be  required  to  notify 
the  appropriate  Federal  funding  agency  of  ap- 
parent cases  of  noncompliance  with  the  Guide- 
lines by  their  grantees.  Finally,  it  should  pre- 


pare an  annual  report  to  Congress  on  the  effec- 
tiveness of  Federal  oversight. 

The  choice  of  an  agency  to  administer  the 
statute  would  be  important.  The  selection  of 
NIH  would  permit  the  use  of  an  existing  admin- 
istrative structure  and  body  of  expertise  and  ex- 
perience. On  the  other  hand,  one  of  the  regu- 
latory agencies  may  take  a more  active  moni- 
toring role  and  be  more  experienced  with 
handling  proprietary  information. 

This  approach  is  similar  to  a bill  introduced  in 
the  96th  Congress,  S.  2234,  but  broader  in 
scope.  The  latter  covered  only  institutions  not 
funded  by  NIH,  and  did  not  contain  provisions 
for  requiring  amendment's  to  the  reports  oi-  foi- 
notifying  other  agencies  of  possible  noncom- 
pliance. The  bill  was  broader  in  one  respect 
because  it  would  have  required  information 
about  prospective  experiments.  This  provision 
had  been  criticized  because  of  the  difficulty  of 
projecting  in  advance  the  course  that  scientific 
inquiry  will  take.  The  goals  of  a monitoring  pro- 
gram can  be  substantially  reached  by  monitor- 
ing ongoing  and  completed  work. 

D.  Congress  could  make  the  NIH  Guidelines  ap- 
plicable to  all  rDNA  work  done  in  the  United 
States. 

The  purpose  of  this  option  is  to  allev  iate  any 
concerns  about  the  effectiveness  of  voluntary 
compliance.  RAC  itself  has  gone  on  record  as 
supporting  mandatory  compliance  with  the 
Guidelines  by  non-NIH  funded  instituions,  in- 
cluding private  companies. 

This  option  has  the  advantages  of  using  an  e,\- 
isting  oversight  mechanism,  which  would  sim- 
ply be  extended  to  industry  and  to  academic  re- 
search funded  by  agencies  other  than  Nlll.  Spe- 
cific requirements  on  technical  (|uestiotis  such 
as  containment  levels,  host-vector  .systems,  and 
laboratory  practices  would  continue  to  he  .set  by 
NIH  in  order  to  accommodate  new  information 
expeditiously;  the  statute  would  simply  codilv 
the  responsibilities  and  proctKlui’es  of  the  cur- 
rent system.  There  would  he  few  transitional 
administrative  problems,  since  tin*  e\p«*rtise 
and  experience  already  exist  at  NIH  However,  it 
would  be  necessary  to  appoint  .several  experts 


Ch.11 — Regulation  of  Genetic  Engineering  • 233 


t 

in  fermentation  and  other  industrial  technolo- 
gies to  RAC  if  production,  as  well  as  research,  is 
to  be  adequately  covered.  In  addition,  the  rec- 
ommendations for  large-scale  containment  pro- 
cedures would  have  to  be  made  part  of  the 
, Guidelines. 

The  major  changes  would  have  to  be  made 
; with  respect  to  enforcement.  Present  penalties 
for  noncompliance— suspension  or  termination 
of  research  funds— are  obviously  inapplicable  to 
industry.  In  addition,  procedures  for  monitor- 
ing compliance  could  be  strengthened.  Some  of 
the  elements  of  option  C could  be  used.  An 
added  or  alternative  approach  would  be  to  in- 
spect facilities. 

The  main  disad\  antage  of  this  option  is  that 
NIH  is  not  a regulatory  agency.  Since  NIH  has 
traditionally  viewed  its  mission  as  promoting 
biomedical  research,  it  would  have  a conflict  of 
interest  between  regulation  and  promotion. 
One  of  the  regulatory  agencies  could  be  given 
the  authority  to  enforce  the  Guidelines  and  to 
adopt  changes  therein.  NIH  could  then  continue 
in  a scientific  advisory  role. 

E.  Congress  could  require  an  environmental  im- 
pact statement  and  agency  approval  before 
any  genetically  engineered  organism  is  inten- 
tionally released  into  the  environment. 

There  have  been  numerous  cases  where  an 
animal  or  plant  species  has  been  introduced  into 
a new  environment  and  has  spread  in  an  uncon- 
trolled and  undersirable  fashion.  One  of  the 
early  fears  about  rDNA  was  that  a new  path- 
ogenic or  otherwise  undesirable  micro-orga- 
nism could  establish  an  environmental  niche. 
Yet  in  pollution  control,  mineral  leaching,  and 
enhanced  oil  recovery,  it  might  be  desirable  to 
release  large  numbers  of  engineered  micro-or- 
ganisms into  the  environment. 

The  Guidelines  currently  prohibit  deliberate 
release  of  any  organism  containing  rDNA  with- 
out approval  by  the  Director  of  NTH  on  advice  of 
RAC.  The  obvious  disadvantage  of  this  prohibi- 
tion is  that  it  lacks  the  force  of  law.  The  release 
of  such  an  organism  without  NIH  approval 
would  be  a prima  facie  case  of  negligence,  if  the 
organism  caused  harm.  However,  it  may  be 
more  desirable  social  policy  to  attempt  to  pre- 


vent this  type  of  harm  through  regulation 
rather  than  to  compensate  for  injuries  through 
lawsuits.  Another  possible  disadvantage  of  the 
present  system  is  that  approval  may  be  granted 
on  a finding  that  the  release  would  present  "no 
significant  risk  to  health  or  the  environment;”  a 
tougher  or  more  specific  standard  than  this  may 
be  desirable. 

A required  study  of  the  possible  consequen- 
ces following  the  release  of  a genetically 
engineered  organism,  especially  a micro-orga- 
nism, would  be  an  important  step  in  ensuring 
safety.  This  option  could  be  implemented  by  re- 
quiring those  proposing  to  release  the  organism 
to  file  an  impact  statement  with  an  agency  such 
as  NIH  or  EPA,  which  would  then  grant  or  deny 
permission  to  release  the  organism.  A disad- 
vantage of  this  option  is  that  companies  and  in- 
dividuals might  be  discouraged  from  developing 
useful  organisms  if  this  process  became  too 
burdensome  and  costly. 

F.  Congress  could  pass  legislation  regulating  all 
types  and  phases  of  genetic  engineering,  from 
research  through  commercial  production. 

The  main  advantage  of  this  option  would  be 
to  deal  comprehensively  and  directly  with  the 
risks  of  novel  molecular  genetic  techniques, 
rather  than  relying  on  the  current  patchwork 
system.  A specific  statute  would  eliminate  the 
uncertainties  over  the  extent  to  which  present 
law  covers  particular  applications  of  genetic  en- 
gineering, such  as  pollution  control,  and  any 
concerns  about  the  effectiveness  of  voluntary 
compliance  with  the  Guidelines. 

Other  molecular  genetic  techniques,  while 
not  as  widely  used  and  effective  as  rDNA,  raise 
similar  concerns.  Of  the  current  techniques,  cell 
fusion  is  the  prime  candidate  for  being  treated 
like  rDNA  in  any  regulatory  framework.  It  per- 
mits the  recombination  of  chromosomes  of 
species  that  do  not  recombine  naturally,  and  it 
may  permit  the  DNA  of  latent  viruses  in  the  cells 
to  recombine  into  harmful  viruses.  No  risk  as- 
sessment of  this  technique  has  been  done,  and 
no  Federal  oversight  exists. 

The  principal  arguments  against  this  option 
are  that  the  current  system  appears  to  be  work- 
ing fairly  well,  and  that  the  limited  risks  of  the 


234  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


techniques  may  not  warrant  the  significantly  in- 
creased regulatory  burden  and  costs  that  would 
result  from  such  legislation.  Congress  will  have 
to  decide  if  that  system  will  remain  adequate  as 
commercial  activity  grows. 

If  Congress  were  to  decide  on  this  option,  the 
legislation  could  incorporate  some  or  all  of  op- 
tions C,  D,  and  E.  The  present  mechanism 
created  by  the  Guidelines  could  be  appropriate- 
ly modified  to  provide  the  regulatory  frame- 
work. The  modifications  could  include  a regis- 
tration and  licensing  system  to  provide  infor- 
mation on  what  work  was  actually  being  done 
and  a means  for  continuous  oversight.  One 
important  type  of  information  would  be  health 
and  safety  statistics  gathered  by  monitoring 
workers  involved  in  the  production  of  products 
from  genetically  engineered  organisms.  Anoth- 
er modification  could  be  a sliding  scale  of 
penalties  for  violations,  ranging  from  monetary 
fines  through  revocation  of  operating  licenses 
to  criminal  penalties  for  extreme  cases. 

It  would  not  be  necessary  to  create  a new 
agency,  which  would  duplicate  some  of  the  re- 
sponsibilties  of  existing  agencies.  Instead,  Con- 
gress could  give  these  agencies  clear  regulatory 
authority  by  amending  the  appropriate  statutes. 
Designating  a lead  agency  would  assure  a more 
uniform  interpretation  and  application  of  the 
laws. 

G.  Congress  could  require  NIH  to  rescind  the 
Guidelines. 

This  option  requires  Congress  to  determine 
that  the  risks  of  rDNA  techniques  are  so  insig- 
nificant that  no  control  or  oversight  is  nec- 
essary. Deregulation  would  have  the  advantage 
of  allowing  funds  and  personnel  currently  in- 
volved in  implementing  the  Guidelines  at  the 
Federal  and  local  levels  to  be  used  for  other  pur- 
poses. In  fiscal  year  1980,  NIH  spent  approxi- 
mately $500,000  in  administering  the  Guide- 
lines; figures  are  not  available  for  the  analogous 
cost  to  academia  and  industry.  Personnel  hours 


spent  have  not  been  estimated.  V'ery  few  people 
work  full-time  on  administering  or  complying 
with  the  Guidelines.  NIH  employs  only  six  peo- 
ple full-time  for  this  purpose,  and  some  institu- 
tions employ  full-time  biological  safety  person- 
nel. However,  over  1,000  people  nationally 
devote  some  effort  to  implementing  the 
Guidelines— members  of  the  IBCs  and  the  scien- 
tists conducting  the  rDNA  experiments  who 
must  take  necessary  steps  to  comply. 

There  are  several  reasons  for  retaining  the 
Guidelines.  First,  sufficient  scientific  concern 
about  risks  exists  for  the  Guidelines  to  prohibit 
certain  experiments  and  require  containment 
for  others.  Second,  they  are  not  particularly 
burdensome,  since  an  estimated  80  to  85  per- 
cent of  all  experiments  can  he  done  at  the 
lowest  containment  levels  and  an  estimated  97 
percent  will  not  require  NIH  approval.  I hird, 
NIH  will  continue  to  serve  an  important  role  in 
continuing  risk  assessments,  in  e\aluating  new 
host-vector  systems,  in  collecting  and  dispersing 
information,  and  in  interpreting  the  Guidelines. 
Fourth,  if  the  Guidelines  were  abolished,  I'cgu- 
latory  activity  at  the  State  and  local  levels  (X)uld 
again  become  actixe.  Finally,  the  oversight  sys- 
tem has  been  flexible  enough  in  the  past  to  lib- 
eralize restrictions  as  ex  idence  indicated  loxxer 
risk. 

H.  Congress  could  consider  the  need  for 
regulating  work  with  all  hazardous  micro- 
organisms and  viruses,  whether  or  not  they 
are  genetically  engineered. 

Micro-organisms  carrying  rDNA,  according  to 
an  increasingly  accepted  xiexv,  represent  just  a 
subset  of  micro-organisms  and  x irus(?s,  xx  hich, 
in  general,  pose  risks.  (d)('  has  puhlish(*d  guide- 
lines for  xvorking  xvith  hazardous  agents  such  as 
polio  virus.  Hoxvever,  such  xvoi'k  is  not  cur- 
rently subject  to  legally  enforceable  l'('deral  reg- 
ulations. It  xvas  not  xvithin  the  scope  of  this 
study  to  examine  this  issue,  hut  it  is  an  emerging 
one  that  Congress  may  xvish  to  consid(>r. 


Chapter  12 

Patenting  Living  Organisms 


chapter  12 


Page 


A Landmark  Decision 237 

Legal  Protection  of  Inventions 237 

Trade  Secrets 237 

Patents 238 

Living  Organisms 239 

The  Chakrabarty  Case 240 

Potential  Impacts  of  the  Decision  and 

Related  Policy  Issues 242 

Impacts  on  Industry 242 

The  Relationship  Between  Patents  and 

Innovation 242 

The  Advantages  of  Patenting  Living 
Organisms 243 


Patent  V.  Trade  Secret  Protection 245 

Impact  of  the  Court's  Decision  on  the 

Biotechnology  Industry 246 

Impacts  of  the  Court’s  Decision  on  the  Patent 
Law  and  the  Patent  and  Trademark  Office  . . 246 
Impact  of  the  Court’s  Decision  on  Academic 

Research 248 

Impacts  of  the  Court’s  Decision  on  Genetic 

Diversity  and  the  Food  Supply 249 

The  Morality  of  Patenting  Living  Organisms.  . . 250 
Private  Ownership  of  Inventions  From 

Publicly  Funded  Research 250 

Issue  and  Options 252 


Chapter  12 

Patenting  Living  Organisms 


A landmark  decision 


In  a 5 to  4 decision  (Diamond  \ . Chakrabartv, 
June  16,  1980),  the  Supreme  Court  ruled  that  a 
manmade  mico-organism  is  patentable  under 
the  current  patent  statutes.  This  decision  was 
alternately  hailed  as  ha\  ing  "assured  this  coun- 
try’s technology'  future”’  and  denounced  as  cre- 
ating “the  Bra\  e New  W orld  that  Aldous  Huxley 
warned  ot.”^  Howe\er,  the  Court  clearly  stated 
that  it  was  undertaking  only  the  narrow  task  of 
determining  u hether  or  not  Congress,  in  enact- 
ing the  patent  statutes,  had  intended  a man- 
made micro-organism  to  be  excluded  from  pat- 
entability solely  because  it  was  ali\  e.  Moreov  er, 
the  opinion  invited  Congress  to  overturn  the 
decision  if  it  disagreed  with  the  Court’s  inter- 
pretation. 


■Prepared  Statement  of  (ienentech.  Inc.,  cited  in  "Science  .Vlav 
Patent  New  Forms  of  Life.  Justices  Rule,  5 to  4.  The  ,Ven  York 
Times.  June  17  1980.  p 1 

'Prepared  statement  of  the  Peoples'  Business  Commission,  cited 
in  "Science  May  Patent  .\ew  Forms  of  Life,  Justices  Rule,  5 to  4,” 
The  .\ew  York  Times,  June  17,  1980,  p.  1. 


Congress  may  want  to  reconsider  the  issue  of 
whether  and  to  what  extent  it  should  specifi- 
cally provide  for  or  prohibit  the  patentability  of 
living  organisms.  While  the  judiciary  operates 
on  a case-by-case  basis,  Congress  can  consider 
all  the  issues  related  to  patentability  at  the  same 
time,  gathering  all  relevant  data  and  taking  tes- 
timony from  the  interested  parties.  The  issues 
involved  go  beyond  the  narrow  ones  of  scien- 
tific capabilities  and  the  legal  interpretations  of 
statutory  wording.  They  require  broader  deci- 
sions based  on  public  policy  and  social  values; 
Congress  has  the  constitutional  authority  to 
make  those  decisions  for  society.  It  can  act  to  re- 
solve the  questions  left  unanswered  by  the 
Court,  ov'errule  the  decision,  or  develop  a com- 
prehensive statutory  approach,  if  necessary. 
Most  importantly.  Congress  can  draw  lines;  it 
can  specifically  decide  which  organisms,  if  any, 
should  be  patentable. 


Legal  protection  of  inventions 


The  inherent  ‘Tight”  of  the  originator  of  a 
new  idea  to  that  idea  is  generally  recognized,  at 
least  to  the  extent  of  deserving  credit  for  it 
when  used  by  others.  At  the  same  time,  it  is  also 
believed  that  worthwhile  ideas  benefit  society 
when  they  are  widely  av  ailable.  Similarly,  when 
an  idea  is  embodied  in  a tangible  form,  sucb  as 
in  a machine  or  industrial  process,  the  inventor 
has  the  "right”  to  its  exclusive  posession  and  use 
simply  by  keeping  it  secret.  However,  if  he  may 
be  induced  to  disclose  the  inv'ention’s  details, 
society  benefits  from  the  new  ideas  embodied 
therein,  since  others  may  build  upon  the  new 
knowledge.  The  legal  system  has  long  recog- 
nized the  competing  interests  of  the  inventor 
and  the  public,  and  has  attempted  to  protect 


both.  The  separate  laws  covering  trade  secrets 
and  patents  are  the  mean  by  which  this  is  done. 

Trade  secrets 

The  body  of  law  governing  trade  secrets  rec- 
ognizes that  harm  has  been  done  to  one  person 
if  another  improperly  obtains  a trade  secret  and 
then  uses  it  personally  or  discloses  it  to  others. 
A trade  secret  is  anything— device,  formula,  or 
information— which  when  used  in  a business 
provides  an  advantage  over  competitors  ig- 
norant of  it— e.g.,  improper  acquisition  includes 
a breach  of  confidence,  a breach  of  a specific 
promise  not  to  disclose,  or  an  outright  theft. 
Trade  secrecy  is  derived  from  the  common  law. 


237 


238  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animais 


as  opposed  to  being  specifically  created  by 
statute;  the  State  courts  recognize  and  protect  it 
as  a form  of  property.  The  underlying  policy  is 
one  of  preventing  unfair  competition  or  unjust 
benefits.  The  protection  lasts  indefinitely.  Two 
well-known  examples  of  long-time  trade  secrets 
are  the  formulas  for  Coca  Cola  and  for  Smith 
Brothers’  black  cough  drops;  the  latter  is  sup- 
posedly over  100  years  old. 

A company  relying  on  trade  secrecy  to  pro- 
tect an  important  invention  must  take  several 
steps  to  effect  that  protection.  These  include; 
permitting  only  key  personnel  to  have  access, 
requiring  such  people  to  sign  complex  contracts 
involving  limitations  on  subsequent  employ- 
ment, and  monitoring  employees  and  com- 
petitors for  possible  breaches  of  security.  Even 
so,  there  are  practical  limitations  to  what  can  be 
done  and  what  can  be  proved  to  the  satisfaction 
of  a court.  Moreover,  independent  discovery  of 
the  secret  by  a competitor  is  not  improper,  in- 
cluding the  discovery  of  a secret  process  by  an 
examination  of  the  commercially  marketed 
product.  Most  importantly,  once  a trade  secret 
becomes  public  through  whatever  means,  it  can 
never  be  recaptured.  Thus,  reliance  on  trade 
secrecy  for  protecting  inventions  can  be  risky. 

Patents 

In  contrast  to  the  common  law  development 
of  trade  secrecy,  patent  law  is  a creation  of  Con- 
gress. The  Federal  patent  statutes  (title  35  of  the 
United  States  Code)  are  derived  from  article  I, 
section  8,  of  the  Constitution,  which  states: 

The  Congress  shall  have  Power  ...  To  pro- 
mote the  Progress  of  Science  and  useful  Arts,  hy 
securing  for  limited  Times  to  Authors  and  In- 
ventors the  exclusive  Right  to  their  respective 
Writings  and  Discoveries. 

This  clause  grants  Congress  the  power  to  cre- 
ate a Federal  statutory  body  of  law  designed  to 
encourage  invention  by  granting  inventors  a 
lawful  monopoly  for  a limited  period  of  time. 
Under  the  current  statutory  arrangement, 
which  is  conceptually  similar  to  the  first  patent 
statutes  promulgated  in  1790,  a patent  gives  the 
inventor  the  right  to  exclude  all  others  from 
making,  using,  or  selling  his  invention  within 


the  United  States  without  his  consent  for  17 
years.  In  return,  the  inventor  must  make  full 
public  disclosure  of  his  invention.  The  policy  be- 
hind the  law  is  twofold.  First,  by  rewarding  suc- 
cessful efforts,  a patent  provides  the  in\  entor 
and  those  who  support  him  with  the  incentixe 
to  risk  time  and  money  in  research  and  develop- 
ment. Second,  and  more  importantly,  the  patent 
system  encourages  public  disclosure  of  techni- 
cal information,  which  may  otherwise  ha\  e re- 
mained secret,  so  others  may  use  the  knowl- 
edge. The  inducement  in  both  cases  is  the  po- 
tential for  economic  gain  through  exploitation 
of  the  limited  monopoly.  Of  coui'se,  there  are 
many  reasons  why  this  potential  may  not  he 
realized,  including  the  existence  of  competing 
products. 

To  qualify  for  patent  protection,  an  imention 
must  meet  three  statutory  reciuirements:  it 
must  be  capable  of  being  classified  as  a process, 
machine,  manufacture,  or  composition  of  mat- 
ter; it  must  he  new,  useful,  and  not  ohx  ious;  and 
it  must  be  disclosed  to  the  |)uhlic  in  sufficient 
detail  to  enable  a person  skilled  in  the  same  oi' 
the  most  closely  related  area  of  tcu’hnolog^v  to 
construct  and  operate  it.  I’lants  that  reproduc<' 
asexually  may  also  he  patented,  hut  slightly  dif- 
ferent criteria  are  used. 

Although  the  categories  in  th(^  first  r'e(|uire- 
ment  are  quite  broad,  th(w  are  not  unlimited.  In 
fact,  the  courts  ha\e  held  such  things  as  scien- 
tific principles,  mathematical  formulas,  and 
products  of  nature  to  he  unpatentahh'  on  th(* 
grounds  that  they  are  only  discox ci  ies  ol  pi«>- 
existing  things— not  the  r(?sult  of  the  inx(’nlix«*, 
creatix'e  action  of  man,  xvhich  is  xx  hat  the  pat»'iit 
laws  are  designed  to  (Micourage.  I his  concept 
was  reaffirmed  in  the  T’/ja/craharfx’ opinion. 

The  recjuirement  that  an  inxcntion  he  uselul. 
new,  and  not  ohxious  further  narmxxs  the 
range  of  patentable  inx (Mitions.  I ’tilitx  I'vists  it 
the  invention  xvorks  and  xxould  hax  «•  .some  bene- 
fit to  society;  the  d(!gr(M’  is  not  import. mt  ,\ox  ci- 
ty signifies  that  tlu;  inxcntion  must  dilhT  I mm 
the  "prior  art”  (publicly  knoxxn  inxentions  or 
knowledge).  Novelty  is  tiot  considered  to 
— e.g.,  if:  1)  the  a[)plicant  for  a patent  is  not 
the  inventor,  2)  the?  inxcntion  xxas  prexiousix 


Ch.  12 — Patenting  Living  Organisms  • 239 


known  or  used  publidv  by  others  in  tbe  United 
States,  or  3)  tbe  invention  was  previously  de- 
scribed in  a U.S.  or  foreign  patent  or  publica- 
tion. rbe  inability  to  meet  the  novelty  require- 
ment is  another  reason  u by  products  of  nature 
are  unpatentable.  \onob\  iousness  refers  to  tbe 
degree  of  difference  between  tbe  in\  ention  and 
the  prior  art.  If  tbe  invention  would  have  been 
obvious  at  tbe  time  it  was  made  to  a person  with 
ordinary  skill  in  that  field  of  technolo^v,  then  it 
is  not  })atentable.  The  policy  behind  tbe  dual 
' criteria  of  nov  eltv  and  nonobv  iousness  is  that  a 
i patent  should  not  take  fi'om  the  public  some- 
' thing  which  it  already  enjoys  or  potentially 
! enjoys  as  an  obv  ious  e.\ tension  of  current 
' knowledge. 

rbe  final  re(|uirement— for  adequate  public 
disclosure  of  an  inv  ention— is  know  n as  the  en- 
I ablement  requirement.  It  is  designed  to  ensure 
I that  the  [)ublic  receives  the  full  benefit  of  the 
I new  knowledge  in  return  for  granting  a limited 
I monopoly.  .As  a public  document,  tbe  patent 
must  contain  a sufficiently  detailed  description 
of  tbe  invention  so  that  others  in  that  field  of 
technolo^v  can  build  and  use  it.  At  the  end  of 
this  description  are  the  claims,  which  define  the 
boundaries  of  the  invention  protected  by  the 
patent. 

The  differences  between  trade  secrets  and 
patents,  therefore,  center  on  the  categories  of 
inventions  protected,  the  term  and  degree  of 
protection,  and  the  disclosure  required.  Only 
those  inventions  meeting  the  statutory  require- 
ments outlined  above  qualify  for  patents  and 
then  only  for  a limited  time,  whereas  anvthing 
giving  an  adv  antage  over  business  competitors 
qualifies  as  a trade  secret  for  an  unlimited  time. 
A patent  requires  full  public  disclosure,  while 
trade  secrecy  requires  an  explicit  and  often 
costly  effort  to  withhold  information.  The  pat- 
ent law  provides  rights  of  exclusion  against 
everyone,  even  subsequent  independent  inven- 
tors, while  the  trade  secrecy  law  protects  only 
against  wrongful  appropriation  of  the  secret. 

Living  organisms 

Although  the  law  for  protecting  inv'entions  is 
[Usually  thought  of  as  applying  to  inanimate  ob- 
jects, it  also  applies  to  certain  living  organisms. 


Any  organism  that  both  meets  the  broad  defini- 
tion of  a trade  secret  and  may  be  lawfully 
owned  by  a private  person  or  entity  can  be  pro- 
tected by  that  body  of  law,  including  micro- 
organisms, plants,  animals,  and  insects.  In  addi- 
tion, plants  are  covered  specifically  by  two  Fed- 
eral statutes,  the  Plant  Patent  Act  of  1930  and 
the  Plant  \ arietv  Protection  Act  of  1970.  Fur- 
thermore, the  Supreme  Court  has  now  ruled 
that  manmade  micro-organisms  are  covered  by 
tbe  patent  statutes.  Its  determination  of  con- 
gressional intent  in  the  Chakrabarty  case  was 
based  significantly  on  an  analysis  of  the  two 
plant  protection  statutes. 

Patent  protection  for  plants  was  not  available 
until  Congress  passed  the  Plant  Patent  Act  of 
1930,  recognizing  that  not  all  plants  were  prod- 
ucts of  nature  because  new  varieties  could  be 
created  by  man.  This  Act  covered  new  and  dis- 
tinct asexually  reproduced  varieties  other  than 
tuber-propagated  plants  or  those  found  in  na- 
ture.* The  requirement  for  asexual  reproduc- 
tion was  based  on  the  belief  that  sexually 
reproduced  varieties  could  not  be  reproduced 
true-to-type  and  that  it  would  be  senseless  to  try 
to  protect  a variety  that  would  change  in  the 
next  generation.  To  deal  with  the  fact  that  or- 
ganisms reproduce,  the  Act  conferred  the  right 
to  exclude  others  from  asexually  reproducing 
the  plant  or  from  using  or  selling  any  plants  so 
reproduced.  It  also  liberalized  the  description 
requirement  for  plants.  Because  of  the  impos- 
sibility of  describing  plants  with  the  same  de- 
gree of  specificity  as  machines,  their  description 
need  only  be  as  complete  as  is  "reasonably  possi- 
ble." 

By  1970,  plant  breeding  technology  had  ad- 
vanced to  where  new,  stable,  and  uniform  vari- 
eties could  be  sexually  reproduced.  As  a result. 
Congress  provided  patent-like  protection  to 
novel  varieties  of  plants  that  reproduced  sexu- 
ally by  passing  the  Plant  Variety  Protection  Act 
of  1970.  Fungi,  bacteria,  and  first-generation 
hybrids  were  excluded.  * * Hybrids  have  a built- 

* Approximately  4,500  plant  patents  have  been  issued  to  date, 
most  for  roses,  apples,  peaches,  and  chrysanthemums. 

•'Originally,  six  v'egetables— okra,  celery,  peppers,  tomatoes, 
carrots,  and  cucumbers— were  also  excluded.  On  Dec.  22,  1980, 
President  Carter  signed  legislation  (H.R.  999)  amending  the  Plant 
\’ariety  Protection  Act  to  include  these  vegetables,  to  extend  tbe 
term  of  protection  to  18  years,  and  to  make  certain  technical 
changes. 


240  • Impacts  of  Applied  Genetics— Micro-Organisms,  Piants,  and  Animais 


in  protection,  since  the  breeder  can  control  the 
inbred,  parental  stocks  and  the  same  hybrid 
cannot  be  reproduced  from  hybrid  seed. 

The  1970  Act,  administered  by  the  Office  of 
Plant  Variety  Protection  within  the  U.S.  Depart- 
ment of  Agriculture  (USDA),  parallels  the  patent 
statutes  to  a large  degree.  Certificates  of  Plant 
V'ariety  Protection  allow  the  breeder  to  exclude 


The  Chakrabarty  case  _ 

In  1972,  Ananda  M.  Chakrabarty,  then  a re- 
search scientist  for  the  General  Electric  Co.,  de- 
veloped a strain  of  bacteria  that  would  degrade 
four  of  the  major  components  of  crude  oil.  He 
did  this  by  taking  plasmids  from  several  dif- 
ferent strains,  each  of  which  gave  the  original 
strain  a natural  ability  to  degrade  one  of  the 
crude  oil  components,  and  putting  them  into  a 
single  strain.  The  new  bacterium  was  designed 
to  be  placed  on  an  oil  spill  to  break  down  the  oil 
into  harmless  products  by  using  it  for  food,  and 
then  to  disappear  when  the  oil  was  gone.  Be- 
cause anyone  could  take  and  reproduce  the  mi- 
crobe once  it  was  used,  Chakrabarty  applied  for 
a patent  on  his  invention.  The  U.S.  Patent  and 
Trademark  Office  granted  a patent  on  the  proc- 
ess by  which  the  bacterium  was  developed  and 
on  a combination  of  a carrier  (such  as  straw) 
and  the  bacteria.  It  refused  to  grant  patent  pro- 
tection on  the  bacterium  itself,  contending  that 
living  organisms  other  than  plant  were  not 
patentable  under  existing  law.  On  appeal,  the 
Court  of  Customs  and  Patent  Appeals  held  that 
the  inventor  of  a genetically  engineered  micro- 
organism whose  invention  otherwise  met  the 
legal  requirements  for  obtaining  a patent  could 
not  be  denied  a patent  solely  because  the  inven- 
tion was  alive.  The  Supreme  Court  affirmed. 

The  majority  opinion  characterized  the  issue 
as  follows:® 

The  question  before  us  in  this  case  is  a nar- 
row one  of  statutory  interpretation  requiring  us 

to  construe  35  U.S.C.  §101,  which  provides: 


^Diamond  v.  Chakrabarty,  100  S.Ct.  2204,  2207  (1980). 


Others  from  selling,  offering  for  sale,  reproduc- 
ing (sexually  or  asexually),  importing,  or  export- 
ing the  protected  variety.  In  addition,  others 
cannot  use  it  to  produce  a hybrid  or  a different 
variety  for  sale.  However,  saving  seed  for  crop 
production  and  for  the  use  and  reproduction  of 
protected  varieties  for  research  is  expressly 
permitted.  The  term  of  protection  is  18  years. 


"Whoever  invents  or  discovers  any  new  and 
useful  process,  machine,  manufacture,  or  com- 
position of  matter,  or  any  new  and  useful  im- 
provement thereof,  may  obtain  a patent  there- 
for, subject  to  the  conditions  and  requirements 
of  this  title.” 

Specifically,  we  must  determine  whether  re- 
spondent’s micro-organism  constitutes  a "manu- 
facture” or  "composition  of  matter”  within  the 
meaning  of  the  statute. 

After  evaluating  the  words  of  the  statute,  the 
policy  behind  the  patent  laws,  and  the  legis- 
lative history  of  section  101  of  the  patent 
statutes  and  of  the  two  plant  pi'otection  Acts, 
the  Court  ruled  that  Congress  had  not  intended 
to  distinguish  between  unpatentable  and  pat- 
entable subject  matter  on  the  basis  of  liv  ing  ver- 
sus nonliving,  but  on  the  basis  of  "pi’oducts  of 
nature,  whether  living  or  not,  and  human-made 
inventions.”'*  Therefore,  the  majority  ruled, 
“[t]he  patentee  has  produced  a new  bacterium 
with  markedly  different  characteilslics  from 
any  found  in  nature  and  one  having  potential 
for  significant  utility.  His  discovery  is  not  na- 
ture’s handiwork,  hut  his  own;  accordingly  it  is 
patentable  subject  matter  under  §101.”®  The 
majority  did  not  see  their  decision  as  extt'nding 
the  limits  of  patentability  beyond  those  set  by 
Congress. 

The  Court  found  that,  in  choosing  such  ex- 
pansive terms  as  "manufactun'”  and  "com- 
position of  matter”— words  that  have  Imumi  in 
every  patent  statute  since  1 793— ('ongress  plain- 
ly intended  the  patent  laws  to  have  a wide 

■■Ibkl,  p.  2,210. 

®lbici,  p.  2,208. 


i 


Ch.12 — Patenting  Living  Organisms  • 241 


scope.  .\loi'eo\  er,  \\  lien  these  law  s wei'e  last  re- 
codified in  1952,  tht'  congressional  committee 
reports  att  irmed  the  intent  of  (Congress  that  pat- 
entable subject  mattei’  "include  an\  thing  under 
the  sun  that  is  made  h\’  man.’'* *^  I'he  ('ourt 
acknow  ledged  that  not  e\  ervthing  is  patentable; 
laws  of  nature,  physical  phenomena,  and 
ahsti'act  ideas  are  not. 

The  ('ourt  founti  the  (io\  ernment’s  argu- 
ments unpersuasi\  e.  S|)ecifically,  that  [lassing 
the  Plant  Patent  .Act  of  1930  and  the  Plant  \ arie- 
ty  Protection  .\ct  of  1970,  which  excluded  bac- 
teria, was  evidence  of  congressional  under- 
standing that  section  101  did  not  apply  to  liv  ing 
organisms:  otherwise:  these  statutes  would 
have  been  unnecessary.  In  disagreeing,  the 
CA)urt  stated  that  the  1930  Act  was  necessary  to 
overcome  the  belief  that  even  artificially  bred 
plants  were  unpatentable  [products  of  nature 
atul  to  relax  the  written  description  require- 
ment, pei  niitting  a description  as  complete  as  is 
"reasonably  possible.”  As  for  the  1970  ,Act,  the 
Court  stated  that  it  had  been  passed  to  extend 
patent-like  protection  to  new  sexually  reproduc- 
ing varieties,  which,  in  1930,  were  believed  to 
he  incapable  of  reproducing  in  a stable,  uniform 
manner.  The  1970  .Act's  exclusion  of  bacteria, 
which  indicated  to  the  Government  that  Con- 
gress had  not  intended  bacteria  to  be  pat- 
entable, was  considered  insignificant  for  a num- 
ber of  reasons. 

The  Gov  ernment  had  also  argued  that  Con- 
gress could  not  have  intended  section  101  to 
cover  genetically  engineered  micro-organisms, 
since  the  technology  was  unforeseen  at  the 
time.  The  majority  responded  that  the  very  pur- 
pose of  the  patent  law  was  to  encourage  new, 
unforeseen  inv  entions,  w hich  was  why  section 
101  was  so  broadly  worded.  Furthermore,  as 
for  the  “gruesome  parade  of  horribles""  that 
might  possibly  be  associated  with  genetic  engi- 
neering, the  Court  stated  that  the  denial  of  a 
patent  on  a micro-organism  might  slow  the  sci- 
entific work  but  certainly  would  not  stop  it;  and 
the  consideration  of  such  issues  involves  policy 
judgments  that  the  legislative  and  executive 

®S.  Rept.  .No.  1979,  82d  Cong.,  2d  sess..  p.  5,  1952:  H.R.  Repl.  N'o. 
1923,  82d  Cong..  2d  sess.,  p.  6.  1952,  cited  in  Diamond  v. 
Chakrabartv.  100  S.Ct.  2204.  2207  (1980). 

• Diamonds.  Chakrabartv,  100  S.Ct.  2204,  2211  (1980). 


branches  of  Government,  and  not  the  courts, 
are  competent  to  make.  It  further  recognized 
that  Congress  could  amend  section  101  to  spe- 
cificallv  exclude  genetically  engineered  orga- 
nisms or  could  write  a statute  specifically  de- 
signed for  them. 

The  dissenting  Justices  agreed  that  the  issue 
was  one  of  statutory  interpretation,  but  inter- 
preted section  101  differently.  They  saw  the 
two  plant  protection  Acts  as  strong  ev  idence  of 
congressional  intent  that  section  101  not  cover 
living  organisms.  In  view  of  this,  the  dissenters 
maintained  that  the  majority  opinion  was  ac- 
tually extending  the  scope  of  the  patent  law's 
beyond  the  limit  set  by  C'ongress. 

rhe  stated  narrowness  of  the  Court’s  decision 
may  limit  its  impact  as  precedent  in  subsequent 
cases  that  raise  similar  issues,  although  not  nec- 
essarily. Certainly,  the  decision  applies  to  any 
genetically  engineered  micro-organism.  It  is  a 
technical  distinction  vvitho(.it  legal  significance 
that  most  of  the  work  being  done  on  such  orga- 
nisms involves  recombinant  DNA  (rDNA)  tech- 
niques, which  Chakrabartv  did  not  use.  The  real 
question  is  whether  or  not  it  would  permit  the 
patenting  of  other  genetically  engineered  or- 
ganisms, such  as  plants,  animals,  and  insects. 
•Any  fears  that  the  decision  might  serve  as  a 
legal  precedent  for  the  patenting  of  human  be- 
ings in  the  distant  future  are  totally  groundless. 
Under  our  legal  system,  the  ow'nership  of  hu- 
mans is  absolutely  prohibited  by  the  13th 
amendment  to  the  Constitution. 

Although  the  Chakrabarty  case  involved  a 
micro-organism,  there  is  no  reason  that  its  ra- 
tionale could  not  be  applied  to  other  organisms. 
In  the  majority’s  view,  the  crucial  test  for  pat- 
entability concerned  whether  or  not  the  micro- 
organism was  manmade.  Conceptually,  there  is 
nothing  in  this  test  that  limits  it  to  micro- 
organisms. The  operative  distinction  is  between 
humanmade  and  naturally  occurring  "things,” 
regardless  of  what  they  are.  Thus,  the  Chakra- 
barty opinion  could  be  read  as  precedent  for  in- 
cluding any  genetically  engineered  organism 
(except  humans)  within  the  scope  of  section  101. 
Whether  a court  in  a subsequent  case  will  inter- 
pret Chakrabarty  broadly  or  narrowly  cannot  be 
predicted. 


242  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


Even  if  section  101  were  interpreted  as  cover- 
ing other  genetically  engineered  organisms, 
they  probably  could  not  be  patented  for  failure 
to  meet  another  requirement  of  the  patent 
laws— the  enablement  requirement.  It  is  gener- 
ally impossible  to  describe  a living  organism  in 
writing  with  enough  detail  so  that  it  can  be 
made  on  the  basis  of  that  description.  Relaxing 
this  requirement  for  plants  was  one  reason 
behind  the  Plant  Patent  Act  of  1930.  For  micro- 
organisms, the  problem  is  solved  by  depositing  a 
publicly  available  culture  with  a recognized  na- 
tional repository  and  referring  to  the  accession 
number  in  tbe  patent.*  While  such  an  approach 

• This  procedui  e was  accepted  hv  the  Court  of  Customs  and  Pat- 
ent Appeals  ICX'PA)  in  upholding  a patent  on  a process  using  micro- 
organisms. Application  of  Argoudelis,  434  F.2d  1390  (CXiPA  1970). 
Phis  procedure  should  also  he  acceptable  for  patents  on  micro- 
organisms themselves. 

Potential  impacts  of  the 
related  policy  issues  

During  the  8-year  history  of  the  Chakrabarty 
case  and  the  surrounding  public  debate,  nu- 
merous assertions  were  made  about  tbe  poten- 
tial impacts  of  permitting  patents  on  genetically 
engineered  organisms.  They  ranged  from  more 
immediate  effects  on  the  biotechnology  indus- 
try, the  patent  system,  and  academic  research 
to  the  long-term  impacts  on  genetic  diversity 
and  the  food  supply.  In  addition,  two  major  pol- 
icy issues  that  have  been  raised  are  the  morality 
of  patenting  living  organisms;  and  the  propriety 
of  permitting  private  ownership  of  inventions 
from  publicly  funded  research. 

Impacts  on  industry 

The  basic  question  for  industry  is  tbe  extent 
to  which  permitting  patents  on  genetically  en- 
gineered organisms  will  stimulate  both  their  de- 
velopment and  the  growth  of  the  industries  em- 
ploying them.  To  ascertain  this  requires  first  an 
examination  of  the  theory  and  social  policies 
underlying  the  patent  system. 


may  be  theoretically  possible  for  animals  and  in- 
sects, it  may  be  logistically  impractical.  How- 
ever, if  tissue  culture  techniques  ad\  ance  to  tbe 
point  where  genetically  engineered  organisms 
can  be  made  from  single  cells  and  stored  indefi- 
nitely in  that  form,  there  appears  to  be  no  rea- 
son to  treat  them  any  differently  than  micro- 
organisms, in  the  absence  of  a specific  statute 
prohibiting  their  patentability. 


decision  and 


THE  RELATIONSHIP  BETWEEN  l»A  TEN  I S 
AND  INNOVATION 

The  patent  system  is  su[)posed  to  stimulate  in- 
novation—tbe  process  by  which  an  iiux'iition  i.s 
brought  into  commercial  use— hecausi*  the*  in- 
ventor does  not  receive  financial  ix'uaixls  until 
the  invention  is  used  commercially.  Ihe  ( on- 
stitution  itself  presumes  this,  as  do  the  statutes 
enacted  pursuant  to  the  |)atent  clause  in  article 
I,  section  8.  Attempts  ha\'e  been  mad(>  to  subject 
this  presumption  to  empirical  analysis;  but  in- 
novation is  extraordinarily  (X)mple\  and  in- 
volves interacting  factor's  that  ai'c  difficult  to 
separate.  In  addition,  the  existence  ol  patent> 
and  trade  secrets  as  altei  iiatix c means  for  |)io- 
tection  makes  it  almost  impossible  to  study  the 
effects  of  patents  alonr'  on  iinx'iition  and  in- 
novation.* 


‘A  major  reason  lor  the  lack  ol  em)iiric.il  sUhIii-s  h.e.  Iiren  Ihe 
lack  ol  a|i|)ro|iriale  data  I he  inlorm.ilioo  .i  \ ail.ihli  - on  Ihr  I’liinh'  i 
ol  palenis  a|)|)lle(l  tor  and  issued  does  not  mdieale  Ihe  ini|“'i  l.n*.  , 
economic  benidits,  or  economic  I'osls  ol  imenlions  Iwhelhi  i I'.e 
ented  or  unpalenledi  dial  ma\  nol  h.i\  e evisled  ,il  ,dl  m m.n  i 
been  crealed  more  slou  K il  nol  lor  ibe  paleni  s\  siem  ' le  I’m 


Ch.  12 — Patenting  Living  Organisms  • 243 


St‘\  ei'al  re’asoiiahle  arguments  ha\  e been  pre- 
sented to  suppor  t the  pi'esum[)tion  that  the  pat- 
ent system  stimulates  innovation.  First,  the  po- 
tential I'oi'  tlie  e.xclusive  eommei'eialization  of  a 
new  pi'ocluet  or-  pi'oeess  ei'eates  the  ineentiv  e to 
undertake  tlie  long,  I'isky,  and  e.xpensive  pi'oc- 
ess  from  I’eseai'ch  thi’ough  de\  elopment  to  mai'- 
keting.  ,\t  evei'v  stage  of  innovation— from  de- 
fining })riorities  and  making  initial  estimates  of 
an  invention  s value  to  advertising  the  finished 
|)i’oduet— the  inventor  and  his  barker's  must 
spend  time,  money,  and  effor  t,  not  onlv  to  de- 
velop a pr'oilirrt  hirt  to  convince  others  of  its 
vvor'th.  Onlv  a small  per'centage  of  new  ideas  or 
inv  entions  sur  v iv  e.  If  a competitor,  particularly 
a lar'ger'  fir'm  with  a well-developed  mar  keting 
cafKihility,  vver'e  free  to  copy  a product  at  this 
point,  smaller'  firms  would  have  little  incentive 
to  r.mdertake  the  pr'ocess  of  inr'rov  ation. 

Second,  the  infor'mation  and  new  knowledge 
disclosed  by  the  patent  allows  others  to  develop 
competing,  and  pr'esirmahly  better,  prodircts  by 
impt'oving  on  the  patented  pt'oduct  or  "in- 
venting ar'ound”  it.  Third,  patents  may  r'edirce 
unnecessat'v  costs  to  individual  firms,  thei'ehy 
freeing  resources  for  firrther  innovation.  Once 
a patent  is  issued,  competitors  can  r'edii'ect 
t'esearch  and  development  (R&.D)  funds  into 
other  at'eas.  For  the  firm  holding  the  patent, 
maintaining  control  over  the  technology  is 
theoretically  less  e.xpensive,  since  the  costs  of 
trade  secret  protection  are  no  longer 
required.  * * 

Anecdotal  accounts  support  the  proposition 
that  patents  stimulate  innovatron;  probably  the 
best  known  is  the  story  of  penicillin.  Although 


dent  Carters  recent  report  on  industrial  innovation,  the  patent 
policy  committee,  composed  of  industry  representatives  having 
long  e.xperience  with  the  patent  system,  recommended  ways  of 
enhancing  inno\ ation  by  impro\ing  the  patent  system,  including 
the  patenting  of  industrially  important  liv  ing  organisms.  However, 
they  pro\  ided  no  hard  economic  data  to  support  their  recommen- 
dation.’ 

“Carole  Kitti,  and  Charles  L.  Trozzo,  The  Effects  of  Patent  and 
Antitrust  Laws,  Regulations,  and  Practices  on  innovation,  vol.  II  (Arl- 
ington, V a..  Institute  for  Defense  Analyses,  1976),  pp.  2,9. 

’L'.S.  Department  of  Commerce,  Advisory  Committee  on  Indus- 
trial Innovation:  Final  Report,  September  1979,  pp.  148-149. 

■ ■ Patent  rights  can  be  very  e.xpensive  to  enforce  against  an  in- 
fringer, howev  er,  should  litigation  be  necessary. 


Sir  .Alexander  Fleming  had  discovered  a prom- 
ising weapon  against  bacterial  infection,  it  took 
him  over  10  years  to  get  the  money  and  facilities 
he  needed  to  pui'ify  and  produce  penicillin  in 
hulk.  Only  W orld  W'ar  II  and  an  international  ef- 
fort finally  accomplished  that  task.  Sir  Howard 
Florey,  who  shared  the  Nobel  prize  with  Flem- 
ing foi'  developing  penicillin,  attributed  the 
delay  to  their  not  having  patented  the  drug, 
vv  hich  he  termed  "a  cardinal  error. 

Some  have  claimed  that  the  monopoly  power 
of  a patent  can  he  used  to  retard  innovation.  A 
corporation  can  legally  refuse  to  license  a pat- 
ent on  a basic  invention  to  holders  of  patents  on 
improvements,  thus  protecting  its  product  from 
becoming  less  attractive  or  obsolete.  On  the 
other  hand,  unless  the  corporation  can  satisfy 
the  market  for  its  product,  it  is  usually  in  its 
economic  interest  to  engage  in  cross-licensing 
arrangements  with  holders  of  improvement  pat- 
ents; it  receives  royalties  and  all  parties  can 
market  the  improved  product.  Cross-licensing 
has  been  misused  several  times  by  a few  domi- 
nant firms  in  an  attempt  to  exclude  innovative 
new  firms  from  their  markets.  Such  arrange- 
ments v iolate  the  antitrust  laws.  W'hether  or  not 
that  body  of  law  adequately  prevents  patent 
misuse  is  beyond  the  scope  of  this  report. 

THE  ADVANTAGES  OF  PATENTING 
LIVING  ORGANISMS 

Given  the  presumed  connection  between 
patents  and  innovation,  the  next  question  is 
whether  patenting  a living  organism  would  add 
significant  protection  for  the  patent  holder,  or 
whether  alternative  approaches  would  be  suffi- 
cient. In  this  context,  it  is  necessary  to  focus  on 
the  present  industrial  applications— which  in- 
volve only  micro-organisms- to  examine  alter- 
native forms  of  patent  coverage  and  to  compare 
the  protection  offered  by  trade  secrecy  with 
that  offered  by  patents. 

Opinions  vary  widely  among  spokesmen  for 
the  genetic  engineering  companies  on  the  value 
of  patenting  micro-organisms.”  Spokesmen  for 
Genentech,  Inc.,  have  stated  numerous  times 


'“Ibid,  pp.  170-171. 

”D.  Dickson,  "Patenting  Living  Organisms:  How  to  Beat  tbe  Bug- 
Rustlers,”  Nature,  vol.  283,  Jan,  10,  1980.  pp.  128-129. 


244  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


that  such  patents  are  crucial  to  the  development 
of  the  industry,  while  others  have  stated  their 
preference  for  trade  secrecy. 

Genentech’s  friend-of-the-court  brief  filed  in 
the  Chakrabarty  case  stated,  ‘The  patent  incen- 
tive did,  and  doubtless  elsewhere  it  will,  prove 
to  be  an  important  if  not  indispensible  factor  in 
attracting  private  support  for  life-giving  re- 
search. Genentech  bas  also  supported  in- 
creased patent  protection  because,  to  attract 
top  scientists  to  the  company,  it  had  to  give 
assurances  that  they  would  be  able  to  publish 
freely.’^  This  severely  curtails  any  reliance  on 
trade  secrets. 

The  rationale  behind  the  contrary  position  is 
based  on  the  belief  that  the  industry  is  moving 
so  quickly  that  today’s  frontrunner  is  not  nec- 
essarily tomorrow’s,  and  that  unique  knowledge 
translates  into  competitive  advantage.  Thus,  in  a 
strategy  similar  to  that  of  the  advanced  micro- 
electronics industry,  firms  may  prefer  to  rely 
on  trade  secrets  even  for  patentable  inventions, 
coupled  with  an  intense  marketing  effort  once 
an  invention  has  reached  the  commercial  stage. 
The  idea  is  to  get  the  jump  on  competitors  and 
to  stay  in  front.*"* 

The  uncertainty  about  whether  micro-orga- 
nisms could  be  patented  before  the  Supreme 
Court's  decision  does  not  appear  to  have  hin- 
dered the  development  of  the  industry.  Clearly, 
companies  did  not  have  any  difficulty  raising 
capital— e.g.,  before  the  decision,  Cetus  Corp. 
had  a paper  value  of  $250  million  without  hold- 
ing a single  patent  on  a genetically  engineered 
organism.  Moreover,  products  such  as  insulin, 
human  growth  hormone,  and  interferon  were 
being  made,  albeit  in  small  quantities,  by  un- 
patented, genetically  modified  organisms.  (See 
ch.  4.) 

Before  the  decision,  companies  relied  either 
entirely  on  trade  secrecy  for  protection,  or  on  a 
combination  of  patents  on  the  microbiological 
process  and  the  product  and  trade  secret  pro- 
tection of  the  mico-organism  itself.  Considering 


‘^Brief  for  Genentech  as  Amicus  Curiae,  p.  3. 

“Thomas  Kiley,  V'ice  President  and  General  Coun.sel  for 
Genentech,  personal  communication,  Apr.  15,  1980. 

'“Dickson,  op  cit.,  p.  128. 


the  existence  of  such  protection,  the  question  is 
what  the  actual  advantages  are  to  patenting  the 
micro-organisms  as  well. 

One  advantage  results  from  the  ahilitv  of  a 
living  organism  to  reproduce  itself.  Dexeloping 
a new  microbe  for  a specific  purpose,  such  as 
the  production  of  human  insulin,  can  be  a long, 
difficult,  and  costly  procedure.  Yet  once  it  is 
developed,  it  reproduces  endlessly,  and  any- 
body acquiring  a culture  would  ha\  e the  benefit 
of  the  development  process  at  little  or  no  cost 
unless  the  organism  were  patented. 

Often,  a company  is  able  to  keep  the  microbe 
a trade  secret,  since  only  the  product  is  sold. 
However,  where  the  microbe  is  the  product- 
such  as  with  Chakrabarty’s  oil-eating  bacteri- 
um-patenting the  organism  is  the  best  means 
of  protection.  Moreox  er,  even  when  a microbe 
itself  can  be  kept  under  lock  and  key,  a com- 
pany desiring  to  patent  the  process  in  which  it  is 
used  must  place  a sample  culture  in  a |)ublic 
repository  to  meet  the  enahlenuMit  rc(|uire- 
ment. 

A conijjetitor  could  legally  obtain  the  mif'ro- 
organism.  If  the  competitor  \\(M'e  to  use*  it  to 
make  the  product  for  commercial  purposes,  the 
company  might  suspect  infi'ingement  but  ba\c 
difficulty  proving  it,  especially  when  the  prod- 
uct is  not  patented.  The  infringing  actixily 
would  take  place  entirely  xvithin  the  confines  of 
the  competitor’s  plant.  M(M’(?  suspicion  is  not  suf- 
ficient legal  grounds  foi’  ins[)ccting  the  com- 
petitor’s plant  for  exidiMice  of  infringement 
when  the  unpatented  product  could  theoret- 
ically be  made  by  many  different  methods 
besides  the  one  patented.  * 

A second,  but  less  (certain,  adxantage  pro- 
vided by  patenting  the  micro-organism  is  that 
even  uses  and  products  of  the  organism  not  dis- 
covered by  the  inx  cMitor  xvould  be  pi-oleeled  in- 
directly. That  is,  xx’hik?  nexv  u.ses  and  products 
could  be  patented  by  their  inx entors,  those  |).it- 
ents  would  be  "dominated"  by  tin*  micro-orga- 
nism patent.  Royalties  xxoiild  haxc  to  be  paid 


'Some  would  ;msucr  this  assei  lion  In  vin  imk  lli.il  a lawMiil 
could  he  slarled  even  on  the  hasis  ol  litlle  ex  ideiu  e the  -iiim*;  ■ ■an 
|)anv  would  r('ly  on  Ihe  di.scoxnx  ptocos  xxhnli  i>  IiImm.iI  .mil 
XX ide-raiif'inf',  lo  prox  Ide  anx'  exisliiif;  ex  idem  e ol  inti  in8''ini'ot 


Ch.12 — Patenting  Living  Organisms  • 245 


\vhene\er  the  micro-organism  was  used  for 
commercial  purposes.  Whether  this  would  he  a 
significant  ad\antage  in  practice  is  uncertain. 
Usually,  onl\’  one  pi'oduct  is  optimally  produced 
hy  a gi\  en  mici'o-organism  and  only  one  micro- 
organism is  best  for  a gi\en  process.  Pre- 
sumably, the  micro-organism’s  inxentor  would 
also  ha\e  disco\ered  and  patented  its  best  use 
and  product. 

.-\nother  alternative  to  patenting  a man-made 
micro-organism,  besides  trade  secrecy,  is  to  pat- 
ent its  manmade  components.  Examples  of 
these  include  a plasmid  containing  the  cloned 
gene,  a sequence  of  D\A,  or  a synthetic  gene 
made  by  the  reverse  transcriptase  process. 
These  components,  which  are  nothing  more 
than  strings  of  inanimate  chemicals,  would  not 
be  unpatentable  products  of  nature  if  they  were 
made  in  the  laboratory  and  were  not  identical  to 
the  natural  material.  Patenting  them  would  not 
be  ecjuivalent  to  patenting  the  entire  organism, 
since  their  function  would  be  affected  in  vary- 
ing degrees  by  the  internal  environment  of  their 
host.  Nevertheless,  the  inventor  of  a partic- 
ularly useful  component,  such  as  an  efficient 
and  stable  plasmid,  might  want  to  patent  it  re- 
gardless of  w hether  or  not  the  organism  could 
be  patented,  since  it  could  be  used  in  an  in- 
definite number  of  different  micro-organisms. 

Thus,  if  Congress  were  to  prohibit  patenting 
of  micro-organisms  because  they  are  alive,  in- 
dustry could  compensate  to  a large  degree  by 
patenting  inanimate  components.  On  the  other 
hand,  if  Congress  allows  the  Supreme  Court’s 
decision  to  stand,  certain  components  will  un- 
doubtedly still  be  patented.  In  fact,  such  patents 
may  become  more  important  than  patents  for 
micro-organisms,  since  the  components  are  the 
critical  elements  of  genetic  engineering. 

PATENT  \ . TR  ADE  SECRET  PROTECTION 

Even  with  the  advantages  provided  by  pat- 
enting a micro-organism,  a company  could  still 
decide  to  rely  on  trade  secrecy.  In  choosing  be- 
tween these  two  options,  it  would  evaluate  the 
following  factors:*^ 


'®R.  Saliwanchik.  '.Microbiological  ln\'entions:  Protect  by  Patent- 
ing or  Maintain  as  a Trade  Secret?"  Developments  in  Industrial 
Microbiology,  \ ol.  19,  1978.  pp.  273,  277. 


• whether  the  organism  itself  or  the  sub- 
stance that  it  makes  will  be  the  commercial 
product, 

• w hether  there  is  any  significant  doubt  of 
its  meeting  the  legal  requirements  for 
patenting, 

• whether  there  is  the  likelihood  of  others 
discovering  it  independently, 

• whether  it  is  a pioneer  invention, 

• what  its  projected  commercial  life  is  and 
how  readily  others  could  improve  on  it  if  it 
were  disclosed  in  a patent, 

• whether  there  are  any  plans  for  scientific 
publication,  and 

• what  the  costs  of  patenting  are  versus  re- 
liance on  trade  secrecy. 

The  first  two  factors  make  the  decision  easy. 
Obviously,  an  organism  like  Chakrabarty’s  can 
best  be  protected  by  a patent.  In  most  instances, 
the  substance  made  by  the  organism  is  the  com- 
mercial product.  In  that  case,  if  there  are  sig- 
nificant doubts  that  the  organism  can  meet  all 
the  legal  requirements  for  patentability,  the 
company  would  probably  decide  to  rely  on 
trade  secrecy. 

The  next  three  factors  require  difficult  de- 
cisions to  be  made  on  the  basis  of  the  charac- 
teristics of  the  new  organism,  its  product,  and 
the  competitive  env  ironment.  If  research  to  de- 
velop a particular  product  is  widespread  and  in- 
tense (as  is  the  case  with  interferon),  the  risk  of 
a competitor  dev^eloping  the  invention  inde- 
pendently provides  a significant  incentive  for 
patenting.  On  the  other  hand,  reverse  engineer- 
ing (examination  of  a product  by  experts  to  dis- 
cover the  process  by  which  it  was  made)  by 
competitors  is  virtually  impossible  for  products 
of  micro-organisms  because  of  the  variability 
and  biochemical  complexity  of  microbiological 
processes. 

Thus,  greater  protection  may  often  lie  in 
keeping  a process  secret,  even  if  the  microbe 
and  the  process  could  be  patented.  This  is  es- 
pecially true  for  a process  that  is  only  a minor 
improvement  in  the  state  of  the  art  or  that  pro- 
duces an  unpatentable  product  already  made  by 
many  competitors.  The  commercial  life  of  the 
process  might  be  limited  if  it  were  patented  be- 
cause infringement  would  be  difficult  to  detect 


246  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


and  not  worth  the  time  and  money  to  prosecute 
Reliance  on  trade  secrecy  might  then  extend  its 
commercial  life. 

Most  companies  would  patent  truly  pioneer 
inventions,  which  often  provide  the  opportunity 
for  developing  large  markets.  Moreover,  pat- 
ents of  this  sort  tend  to  have  long  commercial 
lives,  since  it  is  difficult  to  circumvent  a pioneer 
invention  and  since  any  improvements  are  still 
subject  to  the  pioneer  patent.  Furthermore,  in- 
fringement is  easy  to  detect  because  of  the  in- 
vention’s trailblazing  nature. 

The  last  two  factors  involve  considerations 
secondary  to  a product  and  its  market.  Ob- 
viously, any  publication  of  the  experiments 
leading  to  an  invention  foreclose  the  option  of 
trade  secrecy.  Also,  company  must  evaluate  the 
options  of  protection  via  either  patenting  or 
trade  secrecy  in  terms  of  their  respective  cost 
effectiveness. 

IMPACT  OF  THE  COURT’S  DECISION 
ON  THE  BIOTECHNOLOGY  INDUSTRY 

The  Chakrabarty  decision  will  add  some  pro- 
tection for  microbiological  inventions  by  pro- 
viding companies  with  an  additional  incentive 
for  the  commercial  development  of  their  inven- 
tions, particularly  in  marginal  cases,  by  lower- 
ing uncertainty  and  risk.  A greater  effect  will 
result  from  the  new  information  disclosed  in 
patents  on  inventions  that  otherwise  might  have 
been  kept  secret  indefinitely.  Competitors  and 
academicians  will  gain  new  knowledge  as  well 
as  a new  organism  upon  which  to  build.  The 
Patent  Office  had  deferred  action  on  about  150 
applications,  while  awaiting  the  Court’s  deci- 
sion; as  of  December  1980,  it  was  processing  ap- 
proximately 200  applications  on  micro-orga- 
nisms.'®’* 

Depending  on  the  eventual  number  and  im- 
portance of  patented  inventions  that  would 
have  otherwise  been  kept  as  trade  secrets,  the 
ultimate  effect  of  the  decision  on  innovation  in 
the  biotechnology  industry  could  be  significant. 


"Rene  Tegtmeyer,  Assistant  Cionimissioner  tor  Patents,  U.S.  Pat- 
ent and  Tradeniark  Otfice,  personal  communication,  Jan.  8,  1981. 

‘These  applications  include  about  100  on  geneticallv  en- 
gineered microbes  and  about  100  on  cultures  oT  strains  isolated 
from  nature. 


Conversely,  if  the  Court  had  reached  the  op- 
posite decision,  the  industry  would  ha\e  been 
held  back  only  moderately  because  of  reason- 
ably effective  alternative  means  of  protection. 

Impacts  of  the  Court's  decision  on  the 
patent  law  and  the  Patent 
and  Trademark  Office 

The  key  rationale  supporting  the  Court’s 
holding  Chakrabarty’s  microbe  to  be  patentable 
was  the  fact  that  it  was  manmade;  its  status  as  a 
living  organism  was  irrele\  ant.  The  Patent  Of- 
fice interprets  this  decision  as  also  permitting 
patents  on  micro-organisms  found  in  nature  hut 
whose  useful  properties  depend  on  human  in- 
tervention other  than  genetic  engineering,'^ 
e.g.,  if  the  isolation  of  a pure  culture  of  a 
microbial  strain  induces  it  to  produce  an  an- 
tibiotic, that  pure  culture  would  he  patentahU' 
subject  matter. 

Because  of  the  complexity,  reproducibility, 
and  mutability  of  li\  ing  organisms,  the  dei'ision 
may  cause  some  problems  for  a body  of  law  de- 
signed more  for  inanimate  ohjec'ts  tlian  for  li\  - 
ing  organisms.  It  raises  (luestions  aliout  the 
proper  interpretation  and  application  of  the  re- 
quirements foi'  no\  elty,  nonohx  iousness,  and 
enablement.  In  addition,  it  raises  (|uestions 
about  how  broad  the  scope'  of  patent  coxcrage* 
on  important  mici’o-organisms  should  be'  and 
about  the  continuing  need  lor  tlu'  txxo  plant  jiro- 
tection  Acts.  These  uncertainties  could  I'e'snlt  in 
increased  litigation,  making  it  more'  eliffie  ult  anel 
costly  for  oxvners  eif  pate'nts  ein  lix  ing  eirganisms 
to  enforce  their  rights. 

The  complexity  ejf  living  matte'i'  xxill  make'  it 
difficult  for  anyeine  examining  the'  inxe'iitiein  tei 
determine  if  it  meets  the  reH|uii  e'me'nts  loi-  neix  • 
elty,  nonobviousness,  and  e'liable'me'nt.  Mie  ie)- 
organisms  can  have  differeMit  e'harae  te'ristie-s  in 
different  enviremments.  Meire'eix  e-r,  mie  rohial 
taxonomists  eiften  differ'  ein  the'  pre'e  i.se'  e l.i.s.sil i- 
cation  of  mici’eihial  sti'ains.  Ia  e'ii  at  te'i'  e'xpe'iisix  e' 
tests,  uncertainty  may  still  e'xist  aheiut  xxhe'thi'r 
a specific  micrei-eii’ganism  is  elistine  t lierm  eithe'i' 
known  strains;  scieintists  elei  neit  haxe*  e-omple'le* 


'Mbid,  .liiii.  7,  198  I 


Ch.  12— Patenting  Living  Organisms  • 247 


knowledge  of  an\  single  organism’s  biophysical 
and  biochemical  mechanisms.  C'onsecjuently, 
there  may  he  cases  where  it  is  difficult  to  know 
the  prior  art  precisely  enough  to  make  a deter- 
mination of  no\  elt\ . 

Similarly,  microbial  comple\it\  I'aises  prob- 
lems in  determining  nonoln  iousness  because 
there  are  so  many  different  w ays  of  engineering 
a new  organism  with  a desired  trait— e.g.,  a 
gene  could  he  inserted  into  a given  plasmid  at 
se\  eral  different  positions.  If  a microbe  w ith  the 
gene  at  one  position  in  the  plasmid  were 
patented,  could  a patent  he  denied  to  an  other- 
wise structurally  identical  organism  with  the 
gene  at  a different  position  because  the  second 
was  obvious?  Perhaps  not.  The  second  organism 
w ould  probably  not  be  an  oln  ions  in\  ention  if  it 
pro\ided  significantly  more  of  the  product,  a 
better  quality  product  under  similar  fermenting 
conditions,  or  the  same  product  under  cheaper 
operating  conditions. 

.As  to  enablement,  the  major  problem  has 
been  discussed  pre\  iously:  placing  a culture  of 
the  micro-organism  into  a repository  is  the  ac- 
cepted solution.  One  problem  w ith  repositories, 
howe\  er,  is  their  potential  misuse.  In  a case  in- 
\ ol\  ing  alleged  price  fi.xing  and  unfair  competi- 
tion—e.g.,  the  Federal  Trade  Commission  found 
that  micro-organisms  placed  in  a public  reposi- 
tory pursuant  to  process  and  product  patents 
on  the  antibiotic  .Aureomycin  did  not  produce 
the  antibiotic  in  commercially  significant 
amounts:  in  actual  practice,  other  strains  were 
being  used  for  production,  and  the  company  in- 
\ ol\  ed  was  able  to  benefit  from  a patent,  w'hile, 
in  effect,  retaining  the  crucial  micro-organism 
as  a trade  secret.'®* 

Comple.xitv  also  raises  questions  about  the  ap- 
propriate scope  of  patent  co\  erage.  In  a patent, 
the  in\  entor  is  permitted  to  claim  his  inx  ention 
as  broadly  as  possible,  so  long  as  the  claims 

''‘American  Cyanamid  Co.,  el.  al,  63  FFC  1747,  1905  n.  14  (1963), 
vacated  and  remanded,  363  F.2d  757  (6th  Cir.  1966),  readopted  72 
FTC  623  (1967),  affirmed  401  F.2d  574  (6th  Cir.  1968),  cert,  denied, 
394  L'.S.  920(1969). 

■The  company  had  maintained  that  sec.  112  simply  required  it 
to  deposit  a strain  that  conformed  to  the  description  of  the  one 
found  in  the  patent  application.  Hotve\  er,  it  is  often  the  case  with 
bacteria  that  manv  strains  of  a species  will  conform  to  e\  en  the 
most  precisely  written  description. 


made  do  not  ox'erlap  with  any  "prior  art”  or  ob- 
x’ious  extensions  thereof— e.g.,  a person  who 
dex  eloped  a particular  strain  of  Escherichia  coli 
that  produced  human  insulin  through  a geneti- 
cally modified  plasmid  could  be  entitled  to  a pat- 
ent coxering  all  strains  of  E.  coli  that  produce 
the  insulin  in  the  same  xvay.  Chakrabarty’s  pat- 
ent application— e.g.,  claimed  "a  bacterium  from 
the  genus  Pseudomonas  containing  therein  at 
least  txvo  stable  energy  generating  plasmids, 
each  of  said  plasmids  providing  a separate 
hydrocarbon  degradative  pathway.”  Several 
species  and  hundreds  of  strains  of  Pseudomonas 
fit  this  description.  A patent  limited  to  a par- 
ticular microbial  strain  is  not  particularly 
valuable  because  it  can  easily  be  circumented 
by  applying  the  inventive  concept  to  a sister 
strain;  on  the  other  hand,  a patent  covering  a 
xvhole  genus  of  micro-organism  (or  several)  may 
retard  competition.  This  problem  will  probably 
be  resolx  ed  by  the  Patent  Office  and  the  courts 
on  a case-by-case  basis. 

.Another  aspect  of  the  same  problem  is 
xvhether  a patent  on  an  organism  w^ould  cover 
mutants.  It  xvould  not  if  the  mutation  occurred 
spontaneously  and  sufficiently  altered  the 
claimed  properties.  Hoxvex  er,  if  a nexv  organism 
xvere  made  in  a laboratory  xvith  a patented 
organism  as  a starting  point,  the  situation  xvould 
be  analogous  to  one  xvhere  an  inventor  can  pat- 
ent an  improx  ed  version  of  a machine  but  must 
come  to  terms  xvith  the  holder  of  the  "domi- 
nant” patent  before  marketing  it. 

The  Chakrabarty  decision  also  raises  ques- 
tions about  the  scope  of  section  101  and  its  rela- 
tion to  the  plant  protection  Acts— e.g.,  plant 
tissue  culture  is,  in  effect,  a collection  of  micro- 
organisms; should  it  be  viewed  as  coming  under 
section  101  instead  of  either  of  the  plant  pro- 
tection Acts?  Could  plants  excluded  under  these 
Acts— such  as  tuber-propagated  plants  or  first- 
generation  hybrids— be  patented  under  section 
101?  Could  any  plants  or  seeds  be  patented 
under  section  101,  and  if  so,  is  there  still  a need 
for  the  plant  protection  Acts?  If  there  is  a need, 
xvould  the  Acts  be  administered  better  by  only 
one  agency?  The  Senate  Committee  on  Appro- 
priations has  directed  the  Departments  of  Com- 
merce and  Agriculture  to  submit  a report 


248  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


within  120  days  of  the  Chakrabarty  decision  on 
the  ad\’isabilitv  of  shifting  the  examining  func- 
tion to  USDA.*®  As  of  December  1980,  this  issue 
was  still  under  study.  These  questions  could  be 
resolved  by  the  courts,  but  they  are  probably 
more  amenable  to  a statutory  solution. 

Another  effect  of  the  decision  could  be  on 
patent  enforcement.  The  various  uncertainties 
discussed  above  may  have  to  be  resolved 
through  costly  litigation.  Moreover,  in  specific 
cases,  the  problems  associated  with  describing  a 
micro-organism  in  sufficient  detail  may  increase 
the  chances  that  a patent  will  be  declared 
invalid.  In  any  event,  litigation  costs  would 
probably  increase  as  more  expert  testimony  is 
needed. 

The  fact  that  organisms  mutate  might  intro- 
duce still  another  complication  into  infringe- 
ment actions.  A deposited  micro-organism  is  the 
standard  by  which  possible  infringement  would 
be  judged.  If  it  has  mutated  with  respect  to  one 
of  its  significant  characteristics,  a patent  holder 
who  is  seeking  to  prove  infringement  may  have 
no  case.  While  this  problem  does  not  appear  to 
be  amenable  to  a statutory  solution,  the  risk  of 
such  a mutation  is  actually  quite  small.* 

Because  a living  invention  reproduces  itself, 
the  statutory  definition  of  infringement  may 
have  to  be  changed.  Presently,  infringement 
consists  of  making,  using,  or  selling  a patented 
invention  without  the  permission  of  the  patent 
holder.  Theoretically,  someone  could  take  part 
of  a publicly  available  micro-organism  culture, 
reproduce  it,  and  give  it  away.  Arguably,  this  is 
not  "making”  the  invention,  and  the  patent 
holder  would  have  the  burdensome  and  expen- 
sive task  of  going  after  each  user.  The  two  plant 
protection  statutes  deal  with  this  problem  by 
specifically  prohibiting  unauthorized  repro- 
duction of  the  protected  plant.  This  approach 
may  be  necessary  for  other  living  inventions. 

How  all  of  these  uncertainties  will  affect  the 
Patent  Office’s  processing  of  applications  cannot 
be  predicted.  Currently,  the  average  processing 
time  for  all  applications  is  22  months;  separate 


‘^’S.  Kept.  No.  96-251,  96th  Cong.  1st  sess.,  1979,  p.  46. 

‘Most  micro-organisms  can  be  stored  in  a freeze-dried  form, 
which  entails  virtuallv  no  risk  of  mutation. 


information  on  genetic  engineering  applications 
is  not  available.^®  It  may  take  examiners  longer 
to  process  applications  on  micro-organisms  than 
for  those  covering  only  microbiological  proc- 
esses or  products  because  of  the  interpretixe 
problems  mentioned.  Moreoxer,  the  Patent  Of- 
fice will  have  to  develop  greater  expei'tise  in 
molecular  genetics— a frontier  scientific  field 
that  has  only  recently  been  the  subject  of  patent 
applications.  On  the  other  hand,  the  Office 
generally  faces  this  problem  for  any  nexx-  area 
of  technology. 

In  terms  of  increased  numbers  of  applica- 
tions, the  decision  is  not  expected  to  hax  e a sig- 
nificant effect  on  the  Patent  Office  operations  in 
the  next  few  years.  The  Office  receix  es  appi'ox- 
imately  100,000  applications  a year,  and  it  has 
about  900  examiners,  each  |)rocessing  an  ax  ei'- 
age  of  about  100  applications  per  year.  Kiguix's 
on  the  number  of  apjilications  on  genetically 
engineered  organisms  xary,  depending  on  hoxx- 
the  category  is  defined,  and  precise  information 
has  not  been  tabulated  by  the  Patent  Officiv 
Rough  estimates  indicate  that  in  February  1980 
about  50  applications  xvere  pending,  and  by 
December  1980,  that  numhei'  had  increased  to 
about  100.  Applications  are  being  fiU'd  at  the 
rate  of  about  5 per  month.  Also,  just  oxcm'  100 
are  pending  on  microbes  that  hax  e h(‘en  isolated 
and  purified  from  natui'al  sources,  hut  hax(‘  not 
been  genetically  engineered.  Four  (vxaminers 
are  xvorking  on  both  catc'gories  as  xxcll  as 
others.  Thus,  in  x iexv  of  th(?  total  operations  of 
the  Office,  these  ai)plications  re(|uire  only  a 
small  part  of  its  I’esources.  Ox(M’  the  next  lexx 
years,  the  number  is  (h\pect(ul  to  increa.sc*  be- 
cause of  the  decision  and  dex ('lo|)inents  in  the 
field  but  not  to  a point  xx  Ikm'c  more  than  a fexx 
additional  examiiK'rs  xx  ill  he  need('d.^' 

Impact  of  the  Ccntrt’s  iltu'ision  on 
academic  research 

Many  academicians  haxe  xoiced  concerns 
about  the  effects  on  res(’ai’ch  t)f  the  Chakrahartv 
decision  and  th(^  commercialization  ol  molecu- 
lar biology  in  gtMKM'al.  Fhey  claim  that  the  re- 


r(?gtnicv(M  . |)('i-sr)n;il  (-ommimK  .limn  l)i-i  I '•  I'tsn 
= 'll)i(t.  l)('c  1,5.  1980.  iind  l.in  8 1981 


Ch.  12 — Patenting  Living  Organisms  • 249 


suits  of  rDM.-X  research  are  not  being  published 
while  patent  applications  ai'e  pending,  discus- 
j sion  at  scientific  meetings  is  being  curtailed,  and 
no\el  organisms  are  less  likely  to  he  freely  ex- 
changed. A related  concern  is  that  scientific 
papers  may  not  he  citing  the  work  of  other'  sci- 
entists to  a\oid  casting  doubt  on  the  noxeltv  or 
im  enti\  eness  of  the  author’s  wor  k,  should  he 
decide  to  apply  for  a patent.  Finally,  there  is 
concern  that  the  gi’anting  of  patents  on  basic 
scientific  pr  ocesses  used  in  the  r'esear'ch  lahor'a- 
tor'v  will  dir'ectly  impede  basic  r'esear'ch— e.g., 
two  scientists  ha\e  r-ecently  been  gr'anted  a pat- 
ent on  the  most  fundamental  process  of  molec- 
ular genetic  technologv— the  transfer  of  a gene 
in  a plasmid  using  rDNA  techniques. The  pat- 
ent has  been  tt'ansferr'ed  to  the  uni\ersities 
wher'e  they  did  their  work— Stanford  and  the 
University  of  California  at  San  Fr'ancisco  (L’CSF). 
Although  both  univer'sities  have  stated  they 
would  grant  low-r'oyalty  licenses  to  anyone  who 
complied  with  the  National  Institutes  of  Health 
(NIH)  Guidelines,  subsequent  owners  of  fun- 
damental process  patents  may  not  be  so 
altruistic. 

Thet'e  ar'e  sever'al  r'easons  for  beliex  ing  that 
these  concerns,  although  genirinely  held,  ar'e 
somewhat  overstated.  Fir'st,  patents  on  funda- 
mental scientific  processes  or  organisms  should 
not  directly  hinder  research.  The  courts  ha\e 
interpi'eted  patent  coverage  as  not  applying  to 
r'esearch;  in  other  words,  the  patent  co\  ers  only 
the  commercial  use  of  the  invention. Also,  it 
would  be  difficult  and  prohibitively  expensive 
for  a patent  holder  to  bring  irifringement  ac- 
tions against  a large  number  of  geograpbically 
separated  scientists.  Second,  patents  ultimately 
result  in  full  disclosure.  If  patents  were  not 
available,  trade  secrecy  could  be  relied  on,  with 
tbe  result  that  important  information  might 
never  become  publicly  available.  Third,  al- 
though delays  occur  while  a patent  application 
is  pending,  they  often  happen  anyw'ay  while  ex- 
periments are  being  conducted  or  w'hile  articles 


“L'.S.  Patent  No.  4,237,224,  issued  Dec.  2,  1980. 

“Xaz  Manufacturing  Co.  v.  Chesebrough-Ponds,  Inc.,  211  F.  Supp. 
815  (S.D.X.V.  1962)  (dictum),  affirmed  317  F.2d  679  (2d  Cir.  1963); 
Chesterfield  United  States,  159  F.  Supp.  371  (Ct.  Cl.  1958):  Dugan 
V.  Lear  Avia,  55  F.  Supp.  223  (S.D.X.V . 1944)  (dictum):  Akro  Agate 
Co.  V.  .Master  ,\larble  Co.,  18  F.  Supp.  305  (X'.D.W'.Va.  1937). 


are  being  prepared  for  publication  because  of 
the  competitive  nature  of  modern  science. 

Fssentially,  the  issue  is  the  effect  of  the  com- 
mercialization of  research  results  on  the  re- 
search process  itself.  Even  if  patents  w'ere  not 
available  for  biological  inventions,  tbe  inventor 
would  simply  keep  his  results  secret  if  he  were 
interested  in  commercialization.  V'iewed  from 
this  perspective,  it  is  difficult  to  see  why  the 
availability  of  patents  should  affect  the  ex- 
change of  scientific  information  in  genetic  re- 
search  any  more  than  it  does  in  any  other  field 
of  research  with  commercial  potential.  The 
Chakrabarty  decision  may  inhibit  the  dissemina- 
tion of  information  only  if  it  creates  an  atmos- 
phere that  stimulates  academic  scientists  to 
commercialize  their  findings.  However,  if  it  en- 
courages them  to  rely  on  patents  rather  than  on 
trade  secrets,  it  will  ultimately  enhance  the 
dissemination  of  information. 

Impacts  of  the  Court's  decision  on 
genetic  diversity  and  the  food  supply 

Some  public  interest  groups  have  claimed 
that  patenting  genetically  modified  organisms 
will  adversely  affect  genetic  diversity  and  the 
food  supply.  The  claim  is  based  on  an  analogy  to 
a situation  alleged  to  exist  for  plants.  Briefly,  the 
groups  claim  that  patenting  micro-organisms 
will  irrevocably  lead  to  patents  on  animals, 
which  will  have  the  same  deleterious  effects  on 
the  animal  gene  pool  and  the  livestock  industry 
as  the  tvv'O  plant  protection  Acts  have  had  on  the 
plant  gene  pool  and  the  plant  breeding  industry. 
The  alleged  effects  are:  loss  of  germplasm  re- 
sources as  a result  of  the  elimination  of  thou- 
sands of  varieties  of  plants;  the  increased  risk  of 
widespread  crop  damage  from  pests  and  dis- 
eases because  of  the  genetic  uniformity  result- 
ing from  using  a single  variety;  and  the  increas- 
ing concentration  of  control  of  the  world’s  food 
supply  in  a few  multinational  corporations 
through  their  control  of  plant  breeding  com- 
panies.^'* 

Only  limited  evidence  is  available,  but  no  con- 
clusive connection  has  been  demonstrated  be- 


^‘Brief  for  the  People.s'  Business  Commission  as  Amicus  Curiae, 
pp.  6-13,  Diamond  v.  Chakrabarty,  100  S.  Ct.  2204  (1980). 


250  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


tween  the  plant  protection  laws  and  the  loss  of 
genetic  diversity,  the  encouragement  of  using  a 
single  variety,  and  any  increased  control  by  a 
few  corporations  of  the  food  supply.  (For  a de- 
tailed discussion,  see  ch.  8.)  Therefore,  any  con- 
nection between  patenting  micro-organisms 
and  potential  detrimental  impacts  on  the  live- 
stock industry  appears  tenuous  at  best.  The 
assumptions  that  the  Chakrabarty  decision  will 
inevitably  lead  to  patenting  animals,  and  that 
the  consequences  will  be  tbe  same  as  those 
claimed  to  result  from  granting  limited  owner- 
ship rights  to  varieties  of  plants,  are  speculative. 

The  morality  of  patenting  living 
organisms 

The  moral  issue  is  difficult  to  analyze  because 
it  embodies  at  least  three  overlapping  questions: 
whether  it  is  moral  to  grant  exclusive  rights  of 
ownership  to  a living  species;  whether  patents 
on  lower  forms  of  life  will  inevitably  lead  to 
genetic  engineering  of  humans;  and  whether 
patenting  organisms  undermines  the  generally 
held  belief  in  tbe  uniqueness  and  sanctity  of  life, 
especially  human  life. 

It  is  difficult  to  assess  the  extent  of  the  belief 
that  patenting  living  organisms  is  intrinsically 
immoral,  and  no  such  assessment  has  been 
done.  Its  extent  and  intensity  will  probably  be 
directly  correlated  with  the  complexity  of  the 
organism  involved.  Fewer  people  will  be  dis- 
turbed about  patenting  micro-organisms  than 
about  patenting  cattle.  A belief  in  the  immorali- 
ty of  patenting  a living  organism  is  a value  judg- 
ment to  which  Congress  may  wish  to  give  some 
consideration. 

The  second  aspect  of  the  moral  issue  revolves 
around  the  well-known  metaphor  of  the  “slip- 
pery slope”— the  fear  that  the  first  steps  along 
the  path  of  genetic  engineering  may  irrevocably 
lead  to  man.  Technology,  at  times,  appears  to 
have  its  own  momentum;  the  aphorism  "what 
can  be  done,  will  be  done”  has  been  true  in  the 
past.  Thus,  some  people  fear  that  patenting 
micro-organisms  may  indeed  set  a dangerous 
precedent  and  encourage  the  technology  to  pro- 
gress to  the  point  of  the  ultimate  dehumaniza- 


tion—the  engineering  of  people  as  an  industrial 
enterprise. 

The  Chakrabarty  opinion  was  written  in  nar- 
row terms.  But  while  its  reasoning  might  be  ap- 
plied to  a future  case  involving  an  animal  or  in- 
sect, it  simply  could  not  be  used  to  justify  the 
patenting  of  human  beings  because  of  the  13th 
amendment  to  the  Constitution,  which  prohibits 
the  ownership  of  humans. 

One  way  to  negotiate  the  slippery  slope  is  to 
deal  directly  with  the  adverse  aspects  of  the 
technology.  Barriers  can  be  erected  along  the 
slope;  the  Constitution  already  protects 
humans.  Congress  can  erect  other  harriers  by 
statute,  specifically  drawing  lines  as  to  which 
organisms  can  or  cannot  be  patented. 

The  third  part  of  the  issue  is  religous  or 
philosophical  in  nature.  For  many,  the  patent- 
ing of  a living  organism  undermines  the  awe 
and  deep  respect  they  hold  for  the  uni(|ue  na- 
ture of  life.  Moreover,  it  raises  appi'ehensions  of 
an  ultimate  threat  to  concepts  of  the  nature  of 
humanity  and  its  place  in  the  uni\  erse.  To  th(\se 
people,  if  life  can  be  engineered  and  patented, 
perhaps  it  is  not  special  or  sacred.  If  this  is  ti'ue 
of  lower  organisms,  why  would  human  In'ings 
be  different?  (This  and  other  aspects  of  the 
morality  issue  are  discussed  in  gi’eater  detail  in 
ch.  13.)' 

Private  ownership  of  inventions 
from  publicly  funded  resean'h 

Much  of  the  basic  research  in  molecular’  gr>- 
netics  has  been  funded  by  Federal  gi’ants.  Most 
of  tbe  work  leading  to  the  development  of  I'DN'.A 
techniques— e.g.,  was  performed  at  Stanfoi'd 
University  and  UCSF  under  NIH  grants.  I he 
scientists  involved  have  i’ecei\ed  a patent  on 
that  fundamental  scientific  procrrss.  Sonu*  o[)- 
ponents  of  patenting  oi’ganisms  ha\’e  argued 
that  private  parties  shoitld  not  lu?  per'rnitted  to 
own  inventions  resirlting  fr'om  feder  ally  funded 
R&,D;  and  in  any  evcMit,  th(>r'e  is  something 
special  about  molecular  genetics  that  re(|uirr‘s 
the  Feder'al  Governnumt  to  r’(>tain  ow  rier’ship  o! 


Ch.  12 — Patenting  Living  Organisms  • 251 


federally  funded  in\  entions  and  to  make  them 
generally  a\ailahle  through  none.\clusi\e 
licenses. 

Until  recently,  there  had  been  tio  comprehen- 
si\  e,  gox  ernmentvvide  policy  regarding  owner- 
ship of  patents  on  federally  funded  in\  entions. 
Some  agencies,  such  as  the  Department  of 
Healtli  and  Human  Ser\ices  (DHHS),  [)ermitted 
noii[)i'ofit  institutional  grantees  to  own  [Kitents 
on  inventions  (subject  to  conditions  deemed 
necessary  to  protect  the  public  interest)  if  they 
had  formal  ()i'ocedures  for  administering  them. 
However,  most  agencies  generally  retained  title 
to  such  patents,  making  them  available  to  any- 
one in  the  [)rivate  sector  for  development  and 
possible  commercialization  through  none.x- 
clusiv  e licenses. 

The  rationale  behind  the  policy  was  simply 
that  inventions  developed  hv  public  money 
should  he  av  ailahle  to  all— including  priv  ate  in- 
dustry—on  a tione.vclusive  basis.  This  arrange- 
ment had  been  criticized  as  not  providing  suf- 
ficient incentiv  e for  industry  to  take  the  risks  to 
dev  elop  the  inv  entions.  Of  the  more  than  28,000 
patents  owned  by  the  Government,  less  than  4 
percent  have  been  successfully  licensed;  on  the 
other  hand,  universities,  which  do  grant  ex- 
clusive licenses  on  patents  that  they  own,  have 
been  able  to  license  33  percent  of  their 
patents.-® 

On  December  12,  1980,  President  Carter 
signed  the  Government  Patent  Policy  Act  of 
1980.  The  .Act  sets  forth  congressional  policy 
that  the  patent  system  be  used  to  promote  the 
utilization  of  inv  entions  developed  under  fed- 
erally supported  R&.D  projects  by  nonprofit 
organizations  and  small  businesses.  To  this  end, 
the  organization  or  firm  may  elect  to  retain  title 
to  those  inventions,  subject  to  various  condi- 
tions designed  to  protect  the  public  interest. 
Such  conditions  include  retention  by  the  fund- 
ing agency  of  a nonexclusiv  e,  irrev  ocable,  paid- 
up  license  to  use  the  invention,  and  the  right  of 
the  Government  to  act  where  efforts  are  not 
being  made  to  commercialize  the  invention,  in 
cases  of  health  or  safety  needs,  or  when  the 
use  of  the  inv^ention  is  required  by  Federal  reg- 
ulations. 

2*S.  Rept.  i\o.  96-480,  96th  Cong.  1st  sess,  1979.  p.  2. 


rhere  is  still  the  question  of  whether  patents 
on  molecular  techniques  or  genetically  en- 
gineered micro-organisms  are  sufficiently  dif- 
ferent to  merit  exception  from  any  general  pat- 
ent policy  decided  on  by  Congress.  For  some, 
the  molecular  genetic  techniques  are  unique  be- 
cause they  are  powerful  scientific  tools  that  can 
manipulate  the  life  processes  as  never  before. 
However,  in  a November  1977  report,  NIH  took 
the  following  position  with  regard  to  patents  on 
rDNA  inventions  developed  under  DHHS-NIH 
support:^^* 

There  are  no  compelling  economic,  social,  or 
moral  reasons  to  distinguish  these  inventions 
from  others  involving  biological  substances  or 
processes  that  have  been  patented,  even  when 
partially  or  wholly  developed  with  public  funds. 

The  report  was  prompted  by  the  Stanford- 
L'CSF  patent  application.  Even  though  the  appli- 
cation was  in  accord  with  the  funding  agree- 
ments between  the  institutions  and  NIH,  the 
universities  requested  a formal  NIH  opinion  on 
the  issue  in  view  of  the  intense  public  interest  in 
rDNA  research.  NIH  solicited  comments  from  a 
group  of  approximately  67  individuals,  ranging 
from  academic  and  industrial  scientists  to 
students,  lawyers,  and  philosophers.^®  The 
review'  and  analysis  of  the  responses  were 
referred  to  the  Federal  Interagency  Committee 
on  rDNA  Research,  the  Public  Health  Service, 
and  the  Office  of  the  General  Counsel  of  the  De- 
partment of  Health,  Education,  and  Welfare 
(now  DHHS).  A fairly  uniform  consensus  on  the 
above-quoted  finding  developed  in  this  process; 
the  one  significant  dissenter,  the  Department  of 
Justice,  contended  that  the  Government  should 
retain  ownership  of  any  invention  resulting 
from  federally  funded  rDNA  research  because 
of  the  great  public  interest  in  that  research. 

^’’The  Patenting  of  Recombinant  DNA  Research  Inventions  De- 
veloped under  DHEW  Support:  An  Analysis  by  the  Director,  National 
Institutes  of  Health,  November  1977,  p.  16. 

•The  report  further  concluded  that  no  change  was  necessary  in 
the  basic  NIH  policy  permitting  nonprofit  organizations  to  own 
patents  on  inventions  developed  under  contracts  or  grants  from 
the  Department  of  Health,  Education,  and  Welfare  (now  DHHS), 
subject  to  several  conditions  to  protect  the  public  interest.  The 
only  recommended  change  was  that  the  formal  agreements  be- 
tween NIH  and  the  institutions  be  amended  to  require  that  any 
licensees  of  institutional  patent  holders  comply  with  the  contain- 
ment standards  of  the  NIH  Guidelines  in  any  production  or  use  of 
rDNA  molecules  under  the  license  agreement. 

“Ibid.,  app.  I,  pp.  5-8. 


252  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Issue  and  Options 


ISSUE:  To  what  extent  could  Congress 

provide  for  or  prohibit  the  pat- 
entability of  living  organisms? 

In  its  Chakrabarty  opinion,  the  Supreme 
Court  stated  that  it  was  undertaking  only  the 
narrow  task  of  determining  whether  or  not 
Congress,  in  enacting  the  patent  statutes,  had 
intended  a manmade  micro-organism  to  be  ex- 
cluded from  patentability  solely  because  it  was 
alive.  Moreover,  the  opinion  specifically  invited 
Congress  to  overrule  the  decision  if  it  disagreed 
with  the  Court’s  interpretation. 

Congress  has  several  options.  It  can  act  to  re- 
solve the  questions  left  unanswered  by  the 
Court,  overrule  the  decision,  or  develop  a com- 
prehensive statutory  approach.  Most  important- 
ly, Congress  can  draw  lines;  it  can  decide  which 
organisms,  if  any,  should  be  patentable. 

OPTIONS 

A:  Congress  could  maintain  the  status  quo. 

Congress  could  choose  not  to  address  the 
issue  of  patentability  and  allow  the  law  to  be 
developed  by  the  courts.  The  advantage  of  this 
option  is  that  issues  will  be  addressed  as  they 
arise  in  the  context  of  a tangible,  nonhypo- 
thetical  case.  Some  of  the  issues  raised  in  the 
debate  on  patenting  may  turn  out  to  be  irrel- 
evant as  the  technology  and  the  law  develop. 
Moreover,  many  of  the  uncertainties  raised  by 
the  Chakrabarty  decision  regarding  provisions 
of  the  patent  law  other  than  section  101  may  be 
incapable  of  statutory  resolution.  The  complexi- 
ty of  living  organisms  and  the  increase  in  knowl- 
edge of  molecular  genetics  will  raise  such  broad 
and  varied  questions  that  legal  interpretations 
of  whether  a particular  biological  invention 
meets  the  requirements  of  novelty,  nonobvious- 
ness, and  enablement  will  best  be  done  on  a 
case-by-case  basis  by  the  Patent  Office  and  the 
Federal  courts. 

There  are  two  disadvantages  to  this  option. 
First,  a uniform  body  of  law  may  take  time  to 
develop,  since  judicial  decisions  about  new  legal 
questions  by  different  Federal  courts  may  ini- 


tially conflict.  Second,  the  Federal  judiciary  is 
not  designed  to  take  sufficient  account  of  the 
broader  political  and  social  interests  inx  oK  ed. 

B:  Congress  could  pass  legislation  dealing  with 
the  specific  legal  issues  raised  by  the  Court’s 
decision. 

Many  of  the  legal  questions  do  not  readily 
lend  themselves  to  statutory  resolution.  How- 
ever, three  questions  are  fairly  nai'row  and 
well-defined  and  may  therefore  he  better  re- 
solved by  statute:  1)  Is  there  a continuing  need 
for  the  plant  protection  Acts  if  plants  can  he 
patented  under  section  101?  2)  If  there  is  a con- 
tinuing need  for  these  Acts,  could  they  he  ad- 
ministered better  by  one  agency?  3)  Should  th(> 
definition  of  infringement  he  clarified  by 
amending  section  271  of  the  Federal  Patent 
Statutes  (title  35  LI.S.C.)  to  include  reproduction 
of  a patented  organism  for  the  [jurpose  of  sell- 
ing it? 

Congressional  action  to  clarity  these  issiu's 
would  prox'ide  direction  for  industi'v  and  the 
Patent  Office,  and  it  would  oh\  iate  thi^  need  foi' 
a resolution  through  costly,  time-consuming  lit- 
igation. Lessening  the  chances  of  litigation  or 
the  chances  of  a patent  being  declai’('d  iinalid 
will  provide  some  stimulation  for  innovation  by 
lessening  the  risks  in  commeix'ial  de\  ('lo|)ment. 
In  addition.  Congress  could  determine  that  tlu* 
plant  protection  Acts  could  he  better  admin- 
istered by  one  agency  or  should  he  inc'orporated 
under  the  more  general  pi’cn  isions  of  the  patent 
law;  if  so,  some  administrative*  e.\p(*nse.s  prob- 
ably could  be  saved. 

C;  Congress  could  mandate  a study  of  the  plant 
protection  Acts. 

Two  statutes,  the  Plant  Pate'iit  .Act  of  15)30 
and  the  Plant  \’ai’iety  I’rotection  .\ct  of  15)70. 
grant  ownership  rights  to  plant  breeders  who 
develop  new  and  distinct  varieties  ol  plants 
They  could  serve  as  a model  for  studving  the 
broader,  long-term  pote'iitial  impacts  of  patent- 
ing living  organisms.  An  em|)irical  study  ol  the 
impacts  of  the  plant  protection  laws  h.is  not 
been  done.  Such  a study  would  he  timelv  not 


Ch.12 — Patenting  Living  Organisms  • 253 


only  because  of  the  Chaknibarty  decision,  hut 
also  because  of  allegations  tliat  the  Acts  ha\  e en- 
couraged the  planting  of  uniform  \arieties,  loss 
of  germplasm  resources,  and  inci'eased  concen- 
tration in  the  plant  breeding  industi'v.  In  addi- 
tion, information  about  the  ,-\cts’  affect  on  in- 
no\alion  anti  competition  in  the  breeding  in- 
dustry would  be  relexant  to  this  aspect  of  the 
biotechnologx'  industi’v.  Howex  er,  it  may  be  ex- 
tremely difficult  to  isolate  the  effects  of  these 
laxx  s from  the  effects  of  other  factors. 

D:  Congress  could  prohibit  patents  on  any  living 
organism  or  on  organisnts  other  than  those 
already  subject  to  the  plant  protection  Acts. 

By  prohibiting  patents  on  anx'  lixing  orga- 
nisms, C'ongress  xxould  be  acce[)ting  the 
arguments  of  those  x\  ho  consider  oxx  iiership 
rights  in  lix  ing  organisms  to  be  immoral,  or  xx  ho 
are  concerned  about  other  potentially  adxerse 
impacts  of  such  patents.  Some  of  the  claimed 
impacts  are:  1)  patents  xxould  stimulate  the  de- 
xelopment  of  molecular  genetic  techniques, 
XX  hich  XX  ill  ex  entually  lead  to  human  genetic  en- 
gineering: 2)  patents  contribute  to  an  atmos- 
phere of  increasing  interest  in  commercializa- 
tion, XX  hich  XX  ill  discourage  the  open  exchange 
of  information  crucial  to  scientific  research;  and 
3)  plant  patents  and  protection  certificates  hax  e 
encouraged  the  planting  of  uniform  xarieties, 
loss  of  germplasm  resources,  and  increasing 
concentration  in  the  plant  breeding  industry, 
i .Also,  by  repealing  the  plant  .Acts,  Congress 
!,  xxould  he  rex  ersing  the  policy  determination  it 
i made  in  1930  and  in  1970  that  oxx  nership  rights 
I in  noxel  xarieties  of  plants  xxould  stimulate 
plant  breeding  and  agricultural  innox  ation. 

j A prohibitory  statute  xx  ould  hax  e to  deal  xvith 
those  organisms  at  the  edge  of  life,  such  as 
! xiruses.  .Although  there  are  uncertainties  and 
b disagreements  in  classifying  some  entities  as 
' lix  ing  or  nonlix  ing,  Congress  could  be  arbitrary 
^ in  its  inclusions  and  exclusions,  so  long  as  it 
; clearly  dealt  xx  ith  all  of  the  difficult  cases. 

This  statute  by  itself  xx  ould  slow  but  not  stop 
^ the  dexelopment  of  molecular  genetic  tech- 
niques and  the  biotechnology  industry  because 
! there  are  sex  eral  good  alternatix  es  for  maintain- 
ing e.xclusixe  control  of  biological  inventions: 


maintaining  organisms  as  trade  secrets;  patent- 
ing microbiological  processes  and  their  prod- 
ucts; and  patenting  the  inanimate  components 
of  a genetically  engineered  micro-organism, 
such  as  plasmids,  xvhich  are  the  crucial  ele- 
ments of  the  technique  anyxvav.  The  develop- 
ment xvoLild  be  sloxved  primarily  because  infor- 
mation that  might  otherxvise  become  public 
xxould  be  kept  as  trade  secrets.  A major  conse- 
ciuence  xxould  be  that  desirable  products  xvould 
take  longer  to  reach  the  market.  Also,  certain 
organisms  or  products  that  might  be  marginally 
profitable  yet  beneficial  to  society,  such  as  some 
vaccines,  xvould  be  less  likely  to  be  developed. 
In  such  cases,  the  recovery  of  development 
costs  xvould  be  less  likely  without  a patent  to 
assure  exclusive  marketing  rights. 

Alternatixely,  Congress  could  overrule  the 
Chakrabarty  decision  by  amending  the  patent 
laxv  to  prohibit  patents  on  organisms  other  than 
the  plants  covered  by  the  txvo  statutes  men- 
tioned in  option  C.  This  xvould  demonstrate 
congressional  intent  that  living  organisms  could 
be  patented  only  by  specific  statute  and  alleviate 
concerns  of  those  xvho  fear  the  "slippery  slope.” 

E:  Congress  could  pass  a comprehensive  law 
covering  any  or  all  organisms  (except 
humans). 

This  option  recognizes  the  fact  that  Congress 
can  draw  lines  where  it  sees  fit  in  this  area.  It 
could  specifically  limit  patenting  to  micro-orga- 
nisms or  encourage  the  breeding  of  agricul- 
turally important  animals  by  granting  patent 
rights  to  breeders  of  new  and  distinct  breeds. 
Any  fears  that  such  patents  would  eventually 
lead  to  patents  on  human  beings  would  be  un- 
founded, since  the  13th  amendment  to  the  Con- 
stitution, xvhich  abolished  slavery,  prohibits 
ownership  of  human  life. 

The  statute  would  have  to  define  included  or 
excluded  species  with  precision.  Although  there 
are  taxonomic  uncertainties  in  classifying  or- 
ganisms, Congress  could  arbitrarily  include  or 
exclude  borderline  cases. 

A statute  that  permitted  patents  on  several 
types  of  organisms  could  be  modeled  after  the 
Plant  V^ariety  Protection  Act— e.g.,  it  should 
cover  organisms  that  are  novel,  distinct,  and 


254  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


uniform  in  reproduction;  such  terms  would 
have  to  be  defined.  Infringement  should  include 
the  unauthorized  reproduction  of  the  orga- 
nism—although  reproduction  for  research 
should  be  excluded  to  allow  the  development  of 
new  varieties.  In  fact,  consideration  should  be 
given  to  covering  in  one  statute  plants  and  all 
other  organisms  that  Congress  desires  to  be  pat- 
entable. This  would  provide  the  advantage  of 
comprehensiveness  and  uniform  treatment;  it 
could  also  address  the  problems  discussed 
under  option  B. 

The  impact  of  this  law  cannot  be  assessed 
precisely.  A comprehensive  statute  would  stim- 
ulate the  development  of  new  organisms  and 
their  products  and  would  encourage  dis- 


semination of  technical  information;  however, 
such  a statute  is  not  essential  to  the  de- 
velopment of  the  biotechnology  industry,  since 
incentives  and  alternative  means  for  protection 
already  exist.  The  secondary  impacts  on  society 
of  the  legislation  are  even  harder  to  assess 
because  of  the  scarcity  of  data  from  which  to 
draw  conclusions.  The  policy  judgments  will 
have  to  be  made  by  Congress  after  it  weighs  the 
opinions  of  the  various  interest  groups. 
Through  legislation.  Congress  has  the  chance  to 
balance  competing  views  on  this  controversial 
issue  and,  if  necessary,  to  alleviate  the  primary 
concerns  about  the  long-term  impacts  of  the 
decision— that  higher  organisms  will  inevitably 
be  patented. 


chapter  13 

Genetics  and  Society 


chapter  13 


Page 


Genetics  and  Modern  Science 257 

Special  Problems  Posed  by  Genetics 258 

Science  and  Society 259 


The  "Public”  and  "Public  Participation” 261 

Issues  and  Options 261 

Bibliography;  Suggested  Further  Reading 265 


chapter  13 

Genetics  and  Society 


Genetics  and  modern  science 


In  1979,  tlie  Oi'ganization  tor  Kronomic  C'oop- 
eration  aiul  Hex elopnient  (OK('n)*  published  a 
sur\  py  of  mec'hanisms  for  settling  issues  im  ol\  - 
ing  scienee  and  tec'hnologx-  in  its  member  coun- 
tries.' rbe  ()K(d3  report  noted  tliat:- 

Science  and  technologx’  . . . ha\  e a nunilier  of 
distinguishing  characteristics  w hich  cause  spe- 
cial problems  or  complications.  One  is  ubiciuity: 
they  are  ex  eryu  here.  They  are  at  the  forefront 
of  social  change.  I hey  not  only  ser\  e as  agents 
of  change,  hut  pro\  ide  the  tools  for  analyzing 
social  change.  They  pose,  therefore,  special 
challenges  to  any  society  seeking  to  shape  its 
o\\  n future  and  not  iust  to  react  to  change  or  to 
the  sometimes  undesired  effects  of  change. 

■After  surxeying  member  countries,  OECD 
identified  si.x  factors  that  distinguish  issues  in 
science  and  technologx  from  other  public  con- 
tro\  ersies. 

1.  The  rapidity  of  change  in  science  and  tech- 
nology often  leads  to  concern.  The  science 
of  genetics  is  one  of  the  most  rapidiv  ex- 
panding areas  of  human  know  ledge  in  the 
world  today.  And  the  technology  of  genetics 
is  causing  quick  and  fundamental  changes 
on  a \ariety  of  fronts.  The  news  media 
ha\e  consistently  reported  dexelopments 
in  genetics,  often  with  front-page  stories. 
Consequently,  the  public  has  become  in- 
creasingly aware  of  dex  elopments  in  genet- 
ics and  genetic  technologies  and  the  speed 
with  which  knowledge  in  the  field  is  gath- 
ered and  applied. 

2.  Many  issues  in  today’s  science  and  technol- 
og}’ are  entirely  new.  Protoplast  fusion,  re- 


‘The  members  of  OECD  are:  .Australia,  .Austria,  Belgium, 
Canada.  Denmark.  Finland,  France,  West  Germany,  Greece,  Ice- 
land. Ii'eland,  Italy.  Japan.  Lu.xembourg.  the  Netherlands,  New 
Zealand,  Norwav.  Portugal,  Spain,  Sweden.  Switzerland,  Turkey, 
the  United  Kingdom,  and  the  United  States. 

'Guild  K.  .Nichols,  Technology  on  Trial:  Public  Participation  in  De- 
cision-Making Related  to  Science  and  Technology  (Paris:  Organiza- 
tion for  Economic  Cooperation  and  Dex  elopment.  1979). 

-Ihid..  p.  16. 


comhinant  DNA  (rDNA),  gene  synthesis, 
chimeras,  fertilization  of  mammalian  em- 
hryos  in  \ itro,  and  the  successful  introduc- 
tion of  foreign  genes  into  mammals  were 
the  subjects  of  science  fiction  until  a few 
years  ago.  Now  they  appear  in  newspapers 
and  popular  magazines.  Yet  the  general 
public’s  understanding  of  these  phenom- 
ena is  limited.  It  is  difficult  for  people  to 
exaluate  competing  claims  about  the  dan- 
gers and  benefits  of  this  new'  technology. 

3.  The  scale,  complexity,  and  interdependence 
among  the  technologies  are  greater  than 
people  suspect.  As  in  other  fields,  applica- 
tions of  biological  technology  often  depend 
on  parallel  dex  elopments  in  areas  that  pro- 
\ ide  critical  support  systems.  Breakdowns 
in  these  systems  are  often  as  limiting  as  fail- 
ures in  the  new  technology  itself.  In  other 
parts  of  this  report  for  example,  sophis- 
ticated breeding  systems  in  farm  animals 
and  large-scale  fermentation  processes  for 
single-cell  cultures  are  described.  Besides 
the  biological  technology  required  to  sup- 
port these  systems,  precise  computerized 
operations  are  required  to  ensure  purity, 
safety,  and  process  control  in  fermentation 
and  to  prox'ide  the  population  statistics 
necessary  for  breeding  decisions. 

4.  Some  scientific  and  technological  achieve- 
ments may  be  irreversible  in  their  effects. 
Because  living  organisms  reproduce,  some 
fear  that  it  will  be  impossible  to  contain 
and  control  a genetically  altered  organism 
that  finds  its  way  into  the  environment  and 
produces  undesirable  effects.  Scenarios  of 
escaping  organisms,  pandemics,  and  care- 
less researchers  are  often  draw,m  by  critics 
of  today’s  genetics  research.  The  intention- 
al release  of  recombinant  organisms  into 
the  environment  is  a related  issue  that  will 
need  to  be  resolved  in  the  future. 

Another  example  of  irreversibility, 
brought  about  by  the  demands  placed  on 


257 


258  • Impacts  of  Applied  Genetics— Micro-Organisms,  Piants,  and  Animals 


world  resources,  is  the  accelerating  loss  of 
plant  and  animal  species.  Concern  over  this 
depletion  of  the  world’s  germplasm  arises 
because  genetic  traits  that  might  meet  as 
yet  unknown  needs  are  being  lost. 

5.  There  exist  strong  public  sensibilities  about 
real  or  imagined  threats  to  human  health. 
Mistrust  of  experts  has  been  stimulated  by 
such  events  as  the  accident  at  the  Three- 
Mile  Island  nuclear  plant  and  the  burial  of 
toxic  chemical  wastes  in  the  Love  Canal. 
Regardless  of  the  real  dangers  involved, 
the  public’s  perception  of  danger  can  be  a 
significant  factor  in  decisionmaking.  At 
present,  some  perceive  genetic  technol- 
ogies as  dangerous. 

6.  A challenge  to  deeply  held  social  values  is  be- 
ing raised  by  scientific  and  technological  is- 


sues. The  increasing  control  over  the  inher- 
ited characteristics  of  li\  ing  things  causes 
concern  in  the  minds  of  some  as  to  how 
widely  that  control  should  he  exercised 
and  who  should  be  deciding  about  the 
kinds  of  changes  that  are  made.  Further- 
more, because  genetics  is  basic  to  all  li\  ing 
organisms,  technologies  applicable  to  low- 
er forms  of  life  are  theoretically  applicable 
to  higher  forms  as  well,  including  human 
beings.  Some  wish  to  discourage  applica- 
tions in  lower  animals  because  they  fear 
that  the  use  of  the  technologies  will  pro- 
gress in  increments,  with  more  and  more 
complex  organisms  being  altered,  until  hu- 
man beings  themselves  become  the  object 
of  genetic  manipulation. 


Special  problems  posed  by  genetics 


Genetics  is  just  one  among  several  disciplines 
of  the  biological  sciences  in  which  major  ad- 
vances are  being  made.  Other  areas,  such  as 
neurobiology,  behavior  modification,  and  socio- 
biology, arouse  similar  concerns. 

Genetics  differs  from  the  physical  sciences 
and  engineering  because  of  its  intimate  associa- 
tion with  people.  The  increasing  control  over 
the  characteristics  of  organisms  and  the  poten- 
tial for  altering  inheritance  in  a directed  fashion 
is  causing  many  to  reevaluate  themselves  and 
their  role  in  the  world.  For  some,  this  degree  of 
control  is  a challenge,  for  others,  a threat,  and 
for  still  others,  it  causes  a vague  unease.  Dif- 
ferent groups  have  different  reasons  for  em- 
bracing or  fearing  the  new  genetic  technologies. 
Religious,  political,  and  ethical  reasons  have 
been  advanced  to  support  different  viewpoints. 

The  idea  that  research  in  genetics  may  lead 
some  day  to  the  ability  to  direct  human  evolu- 
tion has  caused  particularly  strong  reactions. 
One  reason  is  that  such  capability  brings  with  it 
responsibility  for  retaining  the  genetic  integrity 
of  people  and  of  the  species  as  a whole,  a re- 
sponsibility formerly  entrusted  to  forces  other 
than  man. 


Others  find  the  idea  of  directing  e\  ()lulion  ex- 
citing. They  view  the  de\elopment  of  g(Mietics 
technologies  in  a positi\e  light,  and  s(>(>  op- 
portunities to  improve  humanity’s  condition. 
They  argue  that  the  capability  to  change  things 
is,  in  fact,  a part  of  evolution. 

Religious  arguments  on  both  sides  of  this 
challenge  have  been  mad(v  I’ojh*  John  I’aul  II 
has  decried  genetic  enginecM’ing  as  running 
counter  to  natural  law.  On  the*  other  hand,  one 
Catholic  |)hilosopher  has  written:’ 

. . . We  have  always  said,  otten  w ithoiit  real 
belief,  that  we  were  and  are  I'realed  by  Led  in 
His  own  image  and  likeness,  l.et  iis  make  m.m 
in  our  image,  after  our  likeness"  logically  means 
that  man  is  by  nature  a creator,  like  bis  ( reator 
Or  at  least  a cocnuitor  in  a very  real,  auesome 
manner.  Not  mere  collaborator,  nor  adminis- 
trator, nor  caretaker.  My  divine  command  we 
are  creators.  V\'by,  then,  sbould  we  be  shocked 
today  to  learn  that  we  can  now  or  soon  w ill  be 
able  to  create  the  man  of  the  futuic’  Why 
should  we  be  horrified  and  denounce  the  sci- 


^Rohert  I'  I'rancociir.  "W  r (an— We  Vtiisl  Rl•lll•^llon^  nn  Itn- 
I'echnolof'ical  Impri'alix'c. ' /'/iro/oi'H a/  Stiulirs  .1.1  .1  st-pli-mlM  i 
1972.  |).  429  and  al  rnninolc  2 


Ch.  13— Genetics  and  Society  • 259 


enlist  or  physician  tor  daring  to  "play  (lod?”  Is  it 
because  we  ha\e  t'orgotten  the  Semitic  (biblical) 
conception  of  creation  as  Clod's  ongoing  col- 
laboration with  man?  Creation  is  our  Clod-gixen 
role,  and  our  task  is  the  ongoing  creation  of  the 
yet  unfinished,  still  e\  ol\  ing  nature  of  man. 

Man  has  played  (lod  in  the  past,  creating  a 
whole  new  artificial  world  for  his  comfort  and 
enjoyment.  ()h\iously  we  ha\e  not  always  dis- 
played the  necessary  wisdom  and  foresight  in 
that  creation:  so  it  seems  to  me  a waste  of  time 
and  energ\’  for  scientists,  ethicists,  and  laymen 
alike  to  heat  their  breasts  today,  continually 
pleading  the  question  of  whether  or  not  we 
have  the  wisdom  to  play  (lod  with  human  na- 
ture and  our  future.  It  is  ohv  ions  we  do  not,  and 
never  will,  have  all  the  foresight  and  prudence 
we  need  for  our  task.  But  I am  also  convinced 
that  a good  deal  of  the  wisdom  we  lack  could 
hav  e been  in  our  hands  if  we  had  taken  serious- 
ly our  human  vocation  as  transcendent  crea- 


Science  and  society  

The  public’s  increasing  concern  about  the  ef- 
fects of  science  and  tecbnoIog\'  has  led  to  de- 
mands for  greater  participation  in  decisions  on 
scientific  and  technological  issues,  not  only  in 
the  United  States  but  throughout  the  world. 
The  demands  imply  new  challenges  to  systems 
of  representative  government;  in  every  West- 
ern country,  new  mechanisms  have  been  de- 
vised for  increasing  citizen  participation.  An  in- 
creasingly informed  population,  skilled  at  exert- 
ing influence  over  policymakers,  seems  to  be  a 
strong  trend  for  the  future.  The  media  has 
played  an  important  role  in  this  development, 
reporting  both  on  breakthroughs  in  science  and 
technologv'  and  on  accidents,  pollution,  and  the 
side-effects  of  some  technologies. 

One  result  has  been  tbe  growing  politiciza- 
tion of  science  and  technology.  VV’hile  perhaps 
misunderstanding  the  nature  of  science  as  a 
process,  the  public  has  become  disenchanted  by 
recent  accidents  associated  with  technology,  by 
experts  who  openly  disagree  with  one  another, 
and  by  the  selective  use  of  information  by  some 
scientific  supporters  to  obtain  a political  objec- 
tive. Tbe  public  has  seen  that  technology  affects 


tures,  creatures  oriented  toward  the  future 
(here  and  hereafter),  a future  in  which  we  are 
cocreators. 

Genetics  thus  poses  social  dilemmas  that  most 
other  technologies  based  in  the  physical  sci- 
ences do  not.  Issues  such  as  sex  selection,  the 
abortion  of  a genetically  defective  fetus,  and  in 
vitro  fertilization  raise  conflicts  between  in- 
dividual rights  and  social  responsibility,  and 
they  challenge  the  religious  or  moral  beliefs  of 
many.  Furthermore,  people  sense  that  genetics 
will  pose  even  more  difficult  dilemmas  in  the  fu- 
ture. Although  many  cannot  fully  articulate  the 
basis  for  their  concern,  considerations  such  as 
those  discussed  in  this  section  are  cited.  The 
strong  emotions  aroused  by  genetics  and  by  tbe 
questions  of  bow  much  and  what  kind  of  re- 
search should  be  done  are  at  least  partly  rooted 
in  deeply  held  human  values. 


the  distribution  of  benefits  in  society;  it  can 
have  unequal  impacts,  and  those  who  pay  or 
w ho  are  most  in  need  are  not  necessarily  always 
those  who  benefit. 

A national  opinion  survey  of  a random  sam- 
ple of  1,679  U.S.  adults  conducted  for  tbe  Na- 
tional Commission  for  tbe  Protection  of  Human 
Subjects  of  Biomedical  and  Behavioral  Research"* 
made  clear  that  there  is  public  doubt  concern- 
ing equity.  Sixty  percent  of  those  polled  felt  that 
new  tests  and  treatments  deriving  from  medical 
research  are  not  equally  accessible  to  all  Amer- 
icans. Seventy  percent  felt  that  those  most  likely 
to  benefit  from  a new  test  or  treatment  of  lim- 
ited availability  were  those  who  could  pay  for  it 
or  w'ho  knew  an  important  doctor.  This  should 
be  compared  with  the  85  percent  who  felt  that  a 
new  test  or  treatment  should  be  available  to 
those  who  apply  first  or  who  are  most  in  need. 


■‘■'Special  Study,  Implications  of  Advances  in  Biomedical  and  Be- 
havioral Research,"  Report  and  Recommendations  of  the  National 
Commission  for  the  Protection  of  Human  Subjects  of  Biomedical 
and  Behavioral  Research,  DHEW  puhlication  No.  (OS)  78-0015. 


260  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Public  concern  and  demand  for  involvement 
in  the  policy  process  is  illustrated  by  the  re- 
sponse of  communities  to  plans  for  laboratories 
that  would  conduct  rDNA  research.  Perhaps  the 
best  known  example  is  Cambridge,  Mass., 
where  plans  were  announced  for  construction 
of  a moderate  containment  laboratory  at  Har- 
vard University.  Concern  over  this  facility  led  to 
the  formation  of  the  Cambridge  Experimenta- 
tion Review  Board  (CERB).  Composed  of  nine  cit- 
izens—all  laymen  with  respect  to  rDNA  re- 
search—the  CERB  spent  6 months  studying  the 
subject  and  listening  to  testimony  from  sci- 
entists with  opposing  points  of  view.  Their  final 
recommendations  did  not  differ  substantially 
from  the  NIH  Guidelines;  hut  the  process  was 
crucial.  CERB  demonstrated  that  citizens  could 
acquire  enough  knowledge  about  a highly  tech- 
nical subject  to  develop  realistic  criteria  and  ap- 
ply them.  Similar  responses  to  proposed  labo- 
ratories have  occurred  in  a number  of  other 
American  communities,  including  Ann  Arbor, 
Mich.,  and  Princeton,  N.J.® 

These  reactions,  and  similar  phenomena  sur- 
rounding controversies  like  nuclear  power,  in- 
dicate that  the  desire  for  citizen  participation  is 
strong  and  widespread.  Recognizing  this,  each 
Federal  agency  has  its  own  rules  and  mech- 
anisms for  citizen  input.  Special  ad  hoc  com- 
missions are  sometimes  formed  to  collect  infor- 
mation from  private  citizens  before  decisions 
are  made  on  particular  projects.  Congressional 
bearings  held  around  the  country  and  in  Wash- 
ington, D.C.,  are  perhaps  the  best  known  of 
these  inquiries.  While  these  mechanisms  some- 
times slow  the  decisionmaking  process,  they 
help  legitimize  some  decisions,  and  their  role 
will  probably  expand  in  the  future. 

In  corporate  science  and  technology,  public 
demands  are  being  felt  as  well.  Present  regula- 
tions for  environmental  protection  and  worker 
and  product  safety  have  significantly  altered 

■'Richard  Hutton,  Bio-Revolution:  DNA  and  the  Ethics  of  Man- 
Made  Life  INew  York:  New  American  l.ibrarv  (Mentor),  19781. 


corporate  research  and  development  efforts. 
The  public  is  also  becoming  more  involved  in 
corporate  decisionmaking— e.g.,  through  '“pub- 
lic accountability”  campaigns  by  stockholders  to 
influence  company  policies. 

With  the  politicization  of  science,  the  process 
of  research  itself  is  coming  under  increasing 
public  scrutiny— most  recently  in  cases  of  possi- 
ble biohazards,  research  with  human  subjects, 
and  research  on  fetuses.  Some  efforts  are  un- 
derway to  require  better  treatment  of  research 
animals  as  well. 

The  relationship  between  science  and  society, 
between  buman  beings  and  tbeir  tools,  is  a con- 
stantly evolving  one.  Tbe  process  that  bas  been 
called  the  "dialogue  within  science  and  tbe  dia- 
logue between  the  scientific  community  ami  tbe 
general  public”®  will  continue  to  search  for 
standards  of  responsibility.  It  is  likely  that  as 
long  as  science  remains  as  dependent  on  public' 
funds  as  it  has  over  the  past  40  years,  it  will  be 
held  accountable  to  public  \ alues.  As  bas  becMi 
noted:^ 

The  technologies  of  war,  industrialization, 
medicine,  environmental  |)rotection,  etc.,  ap- 
pear less  as  the  demonstrations  of  su|)erior 
claims  of  knowledge  and  moi-e  and  more  as  the 
symbols  of  the  ethical  and  political  choices  un- 
derlying the  distrihution  of  the  power  of  scien- 
tific knowledge  among  competing  social  \al- 
ues  ....  This  cultural  shift  of  emphasis  from  the 
role  of  science  in  the  intellectual  construction  of 
reality  to  the  role  of  science;  in  the;  e'thical  con- 
struction of  society  may  indicate  a |)rofound 
transformation  in  the  [)arameters  of  the  social 
assessment  of  science  and  its  relations  to  the  |)o- 
litical  order. 


“Uaniel  Callahan,  "l•.lhil■al  Rc.sponsihilily  in  Si  irtu  c in  Ihr  I ,n  r 
of  tlnccrtain  Cons(‘(|iicncc,s,  ' Ethical  anti  Si  icntifii  Issues  rosetl  In 
Human  Uses  of  Molecular  Genetics,  Marc  I appe  and  Rohrrl  s 
Morison  ((‘d.s.),  , Annals  nl  Ihc  New  N ni  k Acadcim  iil  srirnrrs  Jl,.', 
.Ian.  23,  I97(i,  p.  10. 

'’Yaron  I'./.rahi,  " ( lu>  I’niilics  nl  Ihc  Social  \sscssmrnl  ol  Si  ii'ncc 
in  The  Sot:ial  Assessment  of  Science,  I . Mcndclsnn  I)  \clkm  I' 
Weingart  (cds  ),  Conicrcncc  I'mcccdmgs  (IliciHi-ld  U rsl  (.n 
many:  /\&.W()pitz,  1978),  p 181 


Ch.  13— Genetics  and  Society  • 261 


The  ‘‘public''  and  “public  participation" 


' These  are  terms  with  \astly  ditferent  mean- 
ings to  dit't’erent  people.  Some  take  "the  public” 
to  mean  an  organized  public  interest  gi'oup; 
others  consider  such  groups  the  "professional” 
public  and  feel  thev  ha\e  agendas  that  differ 
I from  those  of  the  less  organized  "general”  pub- 
lic. .\s  OKCD  stated:'* 

I Public  participation  is  a concept  in  search  of  a 
definition.  Because  it  means  different  things  to 
different  people,  agreement  on  what  constitutes 
■ the  public  " and  what  delineates  "'participation'" 

, is  difficult  to  achie\  e.  The  public  is  not  of  course 
homogeneous:  it  is  comprised  of  many  hetero- 
geneous elements,  interests,  and  preoccupa- 
tions. The  emergence  o\er  the  last  several  dec- 
I ades  of  new  and  sometime  \ ocal  special  interest 
groups,  each  with  its  own  set  of  competing 
I claims  and  demands,  attests  to  the  inherent  dif- 
I ficultv  of  achieving  social  and  political  consen- 
sus on  policy  goals  and  programmes  purporting 
to  ser\  e the  common  interest. 


"Nii’hol.s.  op.  I'it..  p.  7. 


Because  publics  differ  with  each  issue,  no  def- 
inition will  be  attempted  here.  It  is  assumed  that 
"the  public”  is  demanding  a greater  role  in  de- 
cisions about  science  and  technology,  and  that  it 
will  continue  to  do  so.  The  different  publics  that 
coalesce  around  different  issues  vary  widely  in 
their  basic  interests,  their  skills,  and  their 
ultimate  objectives.  They  are  the  groups  that 
will  he  heard  in  the  widening  debate  about 
scientific  and  technological  issues,  and  are  part 
of  u'hat  has  been  called  the  "social  system  of 
science.”® 

The  public  has  already  become  involved  in 
the  decisionmaking  process  involving  genetic 
research.  As  the  science  develops,  new  issues  in 
which  .the  public  will  demand  involvement  will 
arise.  The  question  is  therefore:  What  is  the 
best  way  to  involve  the  public  in  decision- 
making? 

'’J.  M.  Ziman.  Public  Knowledge  (Camhridge:  C:anihi'iclge  Univer- 
sity Pre.ss.  1968). 


I 

I Issues  and  Options 


! Three  issues  are  considered.  The  first  is  an 
i issue  of  process,  concerning  public  invoh  ement 
! in  policymaking:  the  second  is  a technical  issue; 
i and  the  third  reflects  the  complexity  of  some 
j issues  associated  with  genetics  that  may  arise  in 
; the  future. 

; ISSUE:  How  should  the  public  he  in- 

, volved  in  determining  policy  re- 

I lated  to  new  applications  of  ge- 

' netics? 

The  question  as  to  whether  the  public  should 
I be  im  oh  ed  is  no  longer  an  issue.  Groups  de- 
mand to  be  involved  when  people  feel  that  their 
interests  are  threatened  in  ways  that  cannot  be 
j resoh  ed  by  representative  democracy. 

I The  more  relevant  questions  are  whether 
f current  mechanisms  are  adequate  to  meet  pub- 
j lie  desires  to  participate  and  whether  a de- 
I 
I 


liberate  effort  should  be  made  to  increase  pub- 
lic knowledge.  The  last  can  only  be  accom- 
plished by  educating  the  public  and  increasing 
its  exposure  both  to  the  issues  and  to  how  peo- 
ple may  be  affected  by  different  decisions. 

OPTIONS: 

A.  Congress  could  specify  that  the  opinion  of  the 
public  must  be  sought  in  formulating  all  major 
policies  concerning  new  applications  of  ge- 
netics, including  decisions  on  funding  of  spe- 
cific research  projects.  A "public  participation 
statement"  could  be  mandated  for  all  such 
decisions. 

B.  Congress  could  maintain  the  status  quo,  allow- 
ing the  public  to  participate  only  when  it  de- 
cides to  do  so  on  its  own  initiative. 

If  option  A were  followed,  there  would  be  no 
cause  for  claiming  that  public  involvement  was 


! 

II 


262  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


inadequate  (as  occurred  after  the  first  set  of 
Guidelines  for  Recombinant  DNA  Research 
were  promulgated).  However,  option  A can  be 
implemented  in  two  ways.  In  the  first,  the  op- 
portunity for  public  involvement  is  always  pro- 
vided, but  need  not  be  taken  if  there  is  no  public 
interest  in  the  topic.  In  the  second,  public  in- 
volvement is  required.  A requirement  for  public 
involvement  would  pose  the  problem  that  if  the 
public  does  not  wish  to  participate  in  a par- 
ticular decision,  then  opinion  will  sometimes  be 
sought  from  an  uninterested  (and  therefore 
probably  uninformed)  public  simply  to  meet  the 
requirement.  Option  A poses  additional  prob- 
lems: What  is  a “major”  policy?  At  what  stage 
would  public  involvement  be  required— only 
when  technological  development  and  applica- 
tion are  imminent  or  at  the  stage  of  basic 
research?  Finally,  it  should  be  noted  that  under 
option  A,  if  the  public’s  contribution  significant- 
ly influences  policy,  the  trend  away  from  deci- 
sionmaking by  elected  representatives  (rep- 
resentative democracy)  and  toward  decision- 
making by  the  people  directly  (“participatory” 
democracy)  may  be  accelerated. 

Option  B would  be  less  cumbersome  and 
would  permit  the  establishment  of  ad  hoc  mech- 
anisms when  necessary.  On  the  other  hand,  by 
the  time  some  issues  are  raised,  strong  vested 
interests  would  already  be  in  place.  The  grow- 
ing role  of  single-issue  advocates  in  U.S.  politics, 
and  their  skill  in  influencing  citizens  and  policy- 
makers, might  abort  certain  scientific  develop- 
ments in  the  future. 

Regardless  of  which  option  is  selected,  it 
would  be  desirable  to  encourage  different 
forms  of  structuring  public  participation  and  to 
evaluate  the  success  of  each  method.  Many  dif- 
ferent approaches  to  public  participation  have 
been  tried  in  the  United  States  and  Western 
Europe  in  attempts  to  resolve  conflicts  over 
science  and  technology.  Some  have  worked  bet- 
ter than  others,  but  most  have  had  rather 
limited  success.  Because  public  demands  for 
involvement  are  not  likely  to  diminish,  the  best 


'"Dorothy  Nelkin  and  Michael  Pollack,  "Pmhlenis  and  Proce- 
dures in  the  Regulation  of  Technological  Risk,"  in  Societal  fUsk  /l.s- 
sessment,  R.  Schwing,  and  W,  Alhers  (eds.)  (New  York:  Plenum 
Pi'ess,  1980). 


ways  to  accommodate  them  need  to  be  iden- 
tified. 

ISSUE:  How  can  the  level  of  public 

knowledge  concerning  genetics 
and  its  potential  be  raised? 

If  public  involvement  is  expected,  an  in- 
formed public  is  clearly  desirable.  Increasing 
the  treatment  of  the  subject,  both  within  and 
outside  the  traditional  educational  system,  is  the 
only  way  to  accomplish  this. 

Within  the  traditional  educational  system,  at 
least  some  educators  feel  that  too  little  tini(>  is 
spent  on  genetics.  Some,  such  as  members  of 
the  Biological  Sciences  Curriculum  Study  Pro- 
gram, are  considering  increasing  the  share  of 
the  curriculum  devoted  to  genetics.  Because* 
science  and  technology  cause  hi'oad  changes  in 
society,  not  only  is  a clearer  perce'ption  of 
genetics  in  particular  needed,  hut  more*  unele*r- 
standing  of  science  in  general.  I'e)r  ahe)ut  half 
the  U.S.  populatiefii,  high  sche)e)l  hie)le)gy  is  the*ir 
last  science  course.  Educate)rs  must  fe)e’us  e)ii 
this  course  to  increase  puhlie*  unele'rstaneling  e>f 
science.  Because  students  ge*neM'ally  finel  pe*e>ple* 
more  interesting  than  rats,  anel  he*e'ause*  human 
genetics  is  a \’ery  [)e)pular  teipie*  in  the*  high 
school  biology  course,  eulue'ateM’s  re)spe)nsihle*  le)r 
the  Biological  Sciences  Uurrie'ulum  Stueh'  Pre)- 
gram  are  considering  ine’reiasing  time*  spe*nt  e>n 
its  study  in  hejpes  e)f  incre*asing  puhlie*  kneew  l- 
edge  not  only  e)f  genetie:s  hut  e>f  se'ie*ne  e*  in  ge*n- 
eral. 

At  the  unixei'sity  lene*l,  me)re*  funels  coulel  he* 
provided  te>  de\e;le)p  e:e)urse?s  e)ii  the*  re*lation- 
ships  between  scie)ne:e),  teu’hne)le)gy,  anel  ,se)e  ie*l\ , 
which  could  he  elesigne*el  he)th  fe)i’  sluele*nts  anel 
for  the  general  puhlie'. 

Several  se)ui'ce;s  e)utsieie*  the*  traelitie)nal  .se  hoe)l 
system  already  we)rk  te>  ineTe*ase*  puhlie’  unele*r- 
standing  of  scieneie)  anel  the*  l■(*latie)nships  he* 
tween  science  anel  se)e:ie*ty.  .Among  lhe*m  are*: 

• Three  pre)grams  ele*\  e*le)|)e*el  by  the*  \alional 
Science  Fe)unelatie)n  le)  improve*  puhlie- 
understaneling  of  anel  iin  e)l\ e*me*nl  in  sci 
ence:  Scienea*  fe)r  the*  Uili/e*n:  Puhlie  I ndi*r 
standing  e>f  Se:ie*ne'e*:  anel  l.lhii’.s  .mel  Values 
in  Se:iene'e*  anel  re*e'hne)le)g\ . 


Ch.  13 — Genetics  and  Society  • 263 


• Science  C'enters  and  similar  projects  spe- 
cifically designed  to  present  science  infor- 
mation in  an  appealing  fashion. 

• New  magazines  that  offei'  science  informa- 
tion to  the  lay  I'eader— another  indication 
of  inci'easing  interest  in  science. 

• Tele\  ision  [)rograms  dealing  w ith  science 
and  technologic.  K.\am[)les  are  the  two  PBS 
series,  \()\  \ and  Cosmos,  and  the  BBC' 
series,  Connections.  (dfS  has  also  hegiin  a 
new  series  called  The  Universe. 

• I ele\  ision  progi'ams  dealing  with  social 
issues  and  \ alue  conflicts.  Pai'ticulai’ly  in- 
teresting is  the  concept  behind  The  Basters. 
In  this  half-hour  j)rime  time  show , the  net- 
work pro\  itles  the  fii'st  half  of  the  show, 
w hich  is  a dramatization  of  a family  in  con- 
flict o\  ei'  a social  or  ethical  issue.  I'he  sec- 
ond half  of  the  show  consists  either  of  a dis- 
cussit)n  about  what  has  been  seen  or  of 
comments  from  people  w ho  call  in. 

One  interesting  possibility  would  he  to  com- 
bine a series  of  Ba.\ter-ty[)e  episodes  on  genetic 
issues  w ith  audience  reaction  using  the  (J,UBE 
s\'stem,  a tw  o-w  ay  cable  telex  ision  system  in 
C'olumhus,  Ohio  (now  e.xpanding  to  other  cities). 
In  this  sx  stem,  telex  ision  x iexx  ers  are  prox  ided 
xxith  a simple  dex  ice  that  enables  them  to 
ansxx  er  questions  asked  ox  er  the  telex  ision.  A 
computer  tabulates  the  responses,  xxhich  can 
either  be  used  by  the  studio  or  immediately 
transmitted  back  to  the  audience.  QUBE  permits 
its  x iexxers  to  do  comparison  shopping  in  dis- 
count stores,  take  college  courses  at  home,  and 
prox  ide  opinion  to  elected  officials.  It  could  be 
effectixely  combined  xx  ith  a program  like  The 
Ba\ters,  to  study  social  issues.  If  sexeral  such 
programs  on  genetics  xx  ere  shoxx  n to  QUBE  sub- 
scribers, audience  learning  and  interest  could 
be  measured. 

Any  efforts  to  increase  public  understanding 
should,  of  course,  be  combined  xx  ith  carefully 
designed  exaluation  studies  so  that  the  effec- 
tix  eness  of  the  program  can  be  assessed. 

OPTIONS: 

A.  Programs  could  be  developed  to  increase 

public  understanding  of  science  and  the  rela- 


tionships between  science,  technology,  and 
society. 

Public  understanding  of  science  in  today’s 
xxorld  is  essential,  and  there  is  concern  about 
the  adequacy  of  the  public’s  knowledge. 

B.  Programs  could  be  established  to  monitor  the 
level  of  public  understanding  of  genetics  and 
of  science  in  general  and  to  determine  whether 
public  concern  with  decisionmaking  in  science 
and  technology  is  increasing. 

Selecting  this  option  xvould  indicate  that 
there  is  need  tor  additional  information,  and 
that  Congress  is  interested  in  involving  the  pub- 
lic in  dex  eloping  science  policy. 

C.  The  copyright  laws  could  be  amended  to  per- 
mit schools  to  videotape  television  programs 
for  educational  purposes. 

Under  current  copyright  law^  videotaping 
telex'ision  programs  as  they  are  being  broadcast 
may  infringe  the  rights  of  the  program’s  owner, 
generally  its  producer.  The  legal  status  of  such 
tapes  is  presently  the  subject  of  litigation.  As  a 
matter  of  policy,  the  Public  Broadcasting  Serv- 
ice negotiates,  xvith  the  producers  of  the  pro- 
grams that  it  broadcasts,  a limited  right  for 
schools  to  tape  the  program  for  educational 
uses.  This  permits  a school  to  keep  the  tape  for  a 
given  period  of  time,  most  often  one  week,  after 
xvhich  it  must  be  erased.  Otherwise,  a school 
must  rent  or  purchase  a copy  of  the  videotape 
from  the  oxvner. 

In  favor  of  this  option,  it  should  be  noted  that 
many  of  the  programs  are  made  at  least  in  part 
xvith  public  funds.  Removing  the  copyright  con- 
straint on  schools  would  make  these  programs 
more  available  for  another  public  good,  educa- 
tion. On  the  other  hand,  this  option  could  have 
significant  economic  consequences  to  the  copy- 
right oxvner,  w'hose  market  is  often  limited  to 
educational  institutions.  An  ad  hoc  committee  of 
producers,  educators,  broadcasters,  and  talent 
unions  is  attempting  to  develop  guidelines  in 
this  area. 


264  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


ISSUE:  Should  Congress  begin  prepar- 

ing now  to  resolve  issues  that 
have  not  yet  aroused  much  pub- 
lic debate  but  that  may  in  the 
future? 

As  scientific  understanding  of  genetics  and 
the  ability  to  manipulate  inherited  characteris- 
tics develop,  society  may  face  some  difficult 
questions  that  could  involve  tradeoffs  between 
individual  freedom  and  societal  need.  This  will 
be  increasingly  the  case  as  genetic  technologies 
are  applied  to  humans.  Developments  are  oc- 
curring rapidly.  Recombinant  DNA  technology 
was  developed  in  the  1970’s.  In  the  spring  of 
1980,  the  first  application  of  gene  replacement 
therapy  in  mammals  succeeded.  Resistance  to 
the  toxic  effect  of  methotrexate,  a drug  used  in 
cancer  chemotherapy,  was  transferred  to  sen- 
sitive mice  by  substituting  the  gene  for  resist- 
ance for  the  sensitive  gene  in  tissue-cultured 
bone  marrow  cells  obtained  from  the  sensitive 
mice.  Transplanted  back  into  the  sensitive  mice, 
the  bone  marrow  cells  now  conferred  resist- 
ance to  the  drug."  In  the  fall  of  1980,  the  first 
gene  substitution  in  humans  was  attempted." 

Although  this  study  was  restricted  to  non- 
human applications,  many  people  assume  from 
the  above  and  other  examples  that  what  can  be 
done  with  lower  animals  can  be  done  with  hu- 
mans, and  will  be.  Therefore,  some  action  might 
be  taken  to  better  prepare  society  for  decisions 
on  the  application  of  genetic  technologies  to 
humans. 

OPTIONS: 

A.  A commission  could  be  established  to  identify 
central  issues,  the  probable  time-frame  for  ap- 
plication of  various  genetic  technologies  to 
humans,  and  the  probable  effects  on  society, 
and  to  suggest  courses  of  action.  The  commis- 
sion might  also  consider  the  related  area  of 
how  participatory  democracy  might  be  com- 
bined with  representative  democracy  in  deci- 
sionmaking. 


"Jean  L.  Marx,  "(iene  I'ransfer  (iiven  a New  I'wist,"  Science 
208:2.'5,  April  1980,  p.  386. 

"(lina  Bari  Kolala  and  Nicholas  Wade,  "Human  Gene  Treatment 
Stirs  New  Debate,"  Sctence  2 10:24,  October  1980,  p.  407. 


B.  The  life  of  the  President's  Commission  for  the 
Study  of  Ethical  Problems  in  Medicine  and 
Biomedical  and  Behavioral  Research  could  be 
emended  for  the  purpose  of  addressing  these 
issues. 

The  11-member  Commission  was  established 
by  Public  Law  95-622  in  November  1978  and 
terminates  on  December  31,  1982.  Its  purpose  is 
to  consider  ethical  and  legal  issues  associated 
with  the  protection  of  human  subjects  in  I'e- 
search;  the  definition  of  death;  and  \'oluntai’v 
testing,  counseling,  information,  and  education 
programs  for  genetic  diseases  as  well  as  any 
other  appropriate  topics  related  to  medicine 
and  to  biomedical  or  heha\  ioral  research. 

In  July  and  September  1980,  the  Cxjmmission 
considered  how  to  respond  to  a statement  from 
the  general  secretaries  of  the  National  (xjuncil 
of  Churches,  the  Synagogue  (Council  of  America, 
and  the  United  States  Catholic  (xjnfei'ence  that 
the  Federal  Government  should  consider  ethical 
issues  raised  by  genetic  engineering.  The  i(>- 
quest  was  prompted  by  the  Su[)r('me  Court  deci- 
sion allowing  patents  on  "new  life  forms.”  I'he 
general  secretaries  stateil  that  "no  gcnernment 
agency  or  committee  is  cui'iTMitly  (‘\('rcising 
adequate  oversight  or  conti’ol,  nor  addressing 
the  fundamental  ethical  (|uestions  (of  geiKMic 
engineering)  in  a major  way,"  and  ask('d  that  the 
President  "provide  a way  for  rc’pi'csentativ cs  of 
a broad  spectrum  of  oui'  society  to  consider 
these  matters  and  acK  ise  the  go\(>rnment  on  its 
necessary  role.”" 

After  testimony  from  \arious  e.\|)erts,  the 
Commission  found  that  the  Go\crnment  is  al- 
ready exercising  ade(|uate  o\  (*rsight  of  the  "bio- 
hazards” associateil  with  I'DNA  research  and  in- 
dustrial production.  The  Commission  decided  to 
prepare  a report  icU'ntifving  \\  hat  are  and  are 
not  realistic  prohUMiis.  II  will  concenlrale  on  the 
ethical  and  social  aspects  of  genetic  lechnolog^\ 
that  are  most  rc^Unant  to  medicine  and  liio- 
medical  research. 

The  Commission  could  he  asked  to  stu(h  the 
areas  it  identifi(\s  and  to  broaden  its  cox  er.ige  to 


"StillCMUMil  by  Ibc  f'cnci  iil  sec  rrl.it  ii-s  I ''  ( .ilhiilH  ( uni,  i 
rncc,  Oi  if’ins.  NC  I loriimrnl.irv  Srr\u  r Mil  III  No  7 liilv  I 
1980. 


Ch.  13 — Genetics  and  Society  • 265 


additional  areas.  This  would  require  that  its 
term  he  e.xtended  and  that  additional  funds  he 
appropriated.  File  Commission  operated  on  $1.2 
million  for  9 months  of  fiscal  vear  1980  and  $1.5 
million  for  fiscal  year  1981.  (a\  en  the  comple.xi- 
ty  of  the  issues  imolved.  the  adequacy  of  this 
le\  el  of  funding  should  he  re\  iew  ed  if  additional 
tasks  are  undei'taken. 

.-\  potential  disad\  antage  of  using  the  existing 
Commission  to  address  societal  issues  associated 
with  genetic  engineering  is  that  a numher  of 
issues  alread\'  exist  and  more  are  likely  to  ap- 
pear in  the  years  ahead.  \ et  there  are  also  other 


issues  in  medicine  and  biomedical  and  be- 
hax'ioral  research  not  associated  with  genetic 
engineering  that  need  review'.  Whether  all 
these  issues  can  be  addressed  by  one  Commis- 
sion should  be  considered.  There  are  obvious 
ad\  antages  and  disadvantages  to  tw'o  Commis- 
sions, one  for  genetic  engineering  and  one  for 
other  issues  associated  w'ith  medicine  and  bio- 
medical and  bebax’ioral  research.  Comments 
from  the  existing  Commission  would  assist  in 
reaching  a decision  on  the  most  appropriate 
course  of  action. 


Bibliography:  suggested  further  reading 


Dobzhanskv,  Theodosiuni,  Genetic  Diversity  and  Hu- 
man Equality  (\ew  \brk:  Basic  Tools,  1973). 

.A  discussion  of  conflicts  between  the  findings 
of  science  and  democratic  social  goals.  Detailed 
coverage  of  the  scientific  basis  for  present  de- 
bates about  intelligence  and  the  misconceptions 
often  in\  oh  ed  in  genetic  \ . en\  ironmental  deter- 
minants of  certain  human  trails. 

Francoeur,  Robert  T.,  "We  Can  - We  Must:  Reflec- 
tions on  the  Technological  Imperative,"  Theologi- 
cal Studies  33  (#3):  428-439,  1972. 

•Argues  that  man  is  a creator  by  virtue  of  his 
special  position  in  nature,  and  that  humans  must 
participate  in  deciding  the  course  of  their  evolu- 
tion. 

Goodfield,  June,  Playing  God:  Genetic  Engineering  and 
the  Manipulation  of  Life  (New  York:  Harper  Col- 
ophon Books,  1977). 

Discusses  the  benefits,  problems  and  potential 
of  genetic  engineering.  Describes  the  moral 
dilemmas  posed  by  the  new  technology.  Suggests 
that  the  ‘social  contract"  between  science  and 
society  is  being  "renegotiated. 

Harmon,  W illis,  An  Incomplete  Guide  to  the  Future 
(New  York:  Simon  and  Schuster,  1976). 

Surveys  how  social  attitudes  and  v'alues  have 
changed  throughout  history  and  how  they  may 
be  changing  today.  Argues  that  mankind  is  in  the 
midst  of  a transition  to  new  values  that  will  affect 
our  w orld  view  as  profoundly  as  did  the  industri- 
al revolution  in  the  19th  century. 


Holton,  Gerald,  and  William  A.  Blanpeid  (eds.).  Sci- 
ence and  Its  Public:  The  Changing  Relationship 
(Boston;  D.  Reidel,  1976). 

A collection  of  essays  on  the  way  science  and 
the  society  of  which  it  is  a part  interact,  and  how 
that  interaction  may  be  changing. 

Hutton,  Richard,  Bio-Revolution:  DNA  and  the  Ethics 
of  Man-Made  Life  (New  York:  New  American  Li- 
brary (Mentor),  1978). 

Reviews  the  history  of  the  debate  about  recom- 
binant DNA,  discusses  the  scientific  basis  for  the 
new  technologies,  and  discusses  the  changing 
relationship  between  science  and  society.  Sug- 
gests how  the  controversies  might  be  resolved. 

Monod,  Jacques,  Chance  and  Necessity  (New  York: 
Alfred  Knopf,  1971). 

A philosophical  essay  on  biology.  Two  seem- 
ingly contradictory  laws  of  science,  the  constan- 
cy of  inheritance  ("necessity”)  and  spontaneous 
mutation  ("chance”)  are  compared  with  more 
vitalistic  and  deontological  views  of  the  universe. 
An  affirmation  of  scientific  knowledge  as  the  on- 
ly "truth”  available  to  man. 

Nichols,  K.  Guild,  Technology  on  Trial:  Public  Participa- 
tion in  Decision-Making  Related  to  Science  and 
Technology  (Paris:  Organization  for  Economic  Co- 
operation and  Development,  1971). 

Reviews  mechanisms  that  have  been  used  by 
countries  in  Europe  and  North  America  to  settle 
disputes  involving  science  and  technology. 


266  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


Sinsheimer,  Robert,  "Two  Lectures  on  Recombinant 
DNA  Research,”  in  The  Recombinant  DNA  Debate, 
D.  Jackson  and  Stephen  Stich  (eds.)  (Englewood 
Cliffs,  N.J.:  Prentice  Hall,  1979),  pp.  85-99. 

Argues  for  proceeding  slowly  and  thoughtfully 
with  genetic  engineering,  for  it  potentially  has 
far-reaching  consequences. 

Tribe,  Laurence,  "Technology  Assessment  and  the 
Fourth  Discontinuity:  The  Limits  of  Instrumental 


Rationality,”  Southern  California  Law  Review  46 
(#3):  617-660,  June  1973. 

An  essay  on  the  fundamental  task  facing  man- 
kind in  the  late  20th  century;  the  problem  of 
choice  of  tools.  New  knowledge,  especially  from 
biology,  will  increasingly  offer  options  for  tech- 
nology, the  use  of  which  will  cause  changes  in 
human  v'alues. 


Appendixes 


I-A.  A Case  Study  of  Acetaminophen  Production 269 

I-B.  A Timetable  for  the  Commercial  Production  of  Compounds  Using 

Genetically  Engineered  Micro-Organisms  in  Biotechnology 275 

I-C.  Chemical  and  Biological  Processes 292 

I- D.  The  Impact  of  Genetics  on  Ethanol— A Case  Study 293 

II- A.  A Case  Study  of  Wheat 304 

II-B.  Genetics  and  the  Forest  Products  Industry  Case  Study 307 

II- C.  Animal  Fertilization  Technologies 309 

III- A.  History  of  the  Recombinant  DNA  Debate 315 

III-B.  Constitutional  Constraints  on  Regulation 320 

III-C.  Information  on  International  Guidelines  for  Recombinant  DNA 322 

r\’.  Planning  Workshop  Participants,  Other  Contractors  and  Contributors,  and 

Acknowledgments 329 


Appendix  I-A 

A Case  Study  of 
Acetaminophen  Production 


Siimmnry 

The  ohjectixf  ot  ihis  rase  study  is  to  demonstrate 
the  eronomir  teasibility  of  a|)[)lying  a f'enetirally 
engineered  strain  to  make  a ihemiral  product  not 
now  produri'd  In  termentation. 

K \(:k(;k()i  \i) 

Acetaminophen  ( \l’  \P)  was  chosen  tor  the  case 
study.  As  an  analf'('sic.  it  lacks  some  ot  the  side  ef- 
fects of  aspirin,  and  is  the  largest  aspirin  substitute 
on  the  market.  .Around  20  million  pounds  (lb)  are 
manufactured  annually.  Mallinckrodt,  Inc.,  produces 
00  to  70  percent:  the  remainder  is  manufactured  f)ri- 
marih  In  ( I’C  International  and  Monsanto  (A).  AP.AP 
is  sold  to  health  care  com[)anies,  \\  hich  market  it  to 
retailers. 

The  .\lc.\eil  (Consumer  Products  di\  ision  of  John- 
son & Johnson,  which  markets  APAP  undei'  the 
trade  name,  lAlenol.  has  the  largest  share  of  the 
market.  ()\er  a dozen  other  com[)anies  in  the  L’nited 
States  sell  it  undei’  other  trade  names. 

One  chemical  manufacturer's  hulk  selling  price  for 
APAP  is  around  S2.05  Ih.'  By  the  time  the  consumer 
purchases  it  at  the  drug  store,  the  markup  results  in 
a selling  price  of  around  S25  to  SoO'lh,  depending  on 
dosage  and  package  sizes.  Thus,  the  total  \ alue  of 
AP  AP  to  the  manufactures  is  some  $50  million  annu- 
ally, w hile  the  total  retail  value  falls  in  the  range  of 
$500  million  to  $1  billion. 

APPROACHES 

• A consen  ative  approach  was  taken,  in  that  only  a 
con\  entional  batch  fermentation  process  was  con- 
sidered. 

• \ ariables  were  selected  pertaining  to  the  choice  of 
the  microbial  pathway:  the  nature  of  the  feed- 
stock: conversion  efficiencies  of  feedstock  to 
.AP.AP:  and  the  final  yield  of  .APAP. 

• Costs  w ere  based  on  proprietary  processes  involv- 
ing startup,  large-scale  fermentation,  and  recovery 
of  APAP. 

• Costs  were  itemized  for  materials  and  supplies: 
labor  distribution:  utilities  (broken  down  by  specif- 
ic energy  requirements  according  to  process  and 
equipment):  equipment  (grouped  according  to 


'('hi'micnl  Markflin^  Hciwrlcr,  .\o\  {>ml)er  and  Decemlier  1979. 


process):  and  building  rec|uirements  (space  needs 
allocated  according  to  pi’ocess). 

CO\CLUSIONS 

• The  [irojecled  cost  for  manufacturing  APAP  by 
means  of  batch  fei’inentation,  using  a genetically 
engineeretl  sti  ain,  amounts  to  $l.()5/lb.  Phis  cost  is 
ba.sed  on  a plant  [u-oducing  10  million  lb  of  APAP 
annually. 

• .As  a rule  of  tbumb,  the  gross  margin  for  manufac- 
ture of  a chemical  such  as  APAP  should  approx- 
imate 50  [lercent  of  sales,  '('he  gross  margin  repre- 
sents the  |)i'ofit  before  general  and  administrative, 
marketing  and  selling,  and  research  and  develop- 
ment expenses.  Tbe  gross  margin  for  all  of  the 
products  made  by  Mallinckrodt,  the  largest  man- 
ufacturer of  AP.AP,  amounted  to  39  and  37  percent 
of  sales  in  1977  and  1978,  respectively.^  The  gross 
margin  foi'  Monsanto,  a much  larger  company 
than  .Mallinckrodt  but  a smaller  manufacturer  of 
AP.AP,  amounted  to  27  and  26  percent  of  all  sales 
in  1976  and  1977,  respectively.^  If  the  gross  mar- 
gin for  APAP  is  as  high  as  50  percent  of  sales,  its 
current  cost  of  manufacture  should  amount  to 
$1,325/11),  based  on  a bulk  selling  price  of  $2. 65/lb. 
Therefore,  its  projected  cost  when  produced  by 
fermentation  is  around  20  percent  lower  than  its 
estimated  cost  udien  produced  by  chemical  syn- 
thesis. 

• If  the  selling  price  of  APAP  produced  by  fermenta- 
tion is  marked  up  100  percent,  the  bulk  selling 
price  becomes  $2. 10/lb.  This  decrease  of  $0. 55/lb 
could  be  transformed  into  cost  savings  of  around 
$5  to  $10/lb  to  the  consumer.  These  economies 
would  result  in  an  annual  cost  saving  to  the  con- 
sumer of  $100  million  to  $200  million. 

• Current  processes  for  synthesizing  APAP  from 
nitrobenzene  do  not  appear  to  pose  significant 
pollution  problems,  although  a number  of  side 
products  are  formed  and  must  be  removed.”  ® ® 
Howev'er,  a fermentation  process  would  be  ev'en 

^Mallinckrodl,  \m-..  Annual  Heport,  1978. 

■'Monsanto  Co.,  Annual  Rcporl,  1977. 

•*H.  C.  Benner,  "Proress  for  i’repariiif'  .Xminoplienol,"  U.S.  Patent 
;i,;i83.4i(;.  i968. 

■9\  ,\.  Baron,  R.  C.  Benner,  and  ,\.  f,.  Weinherg,  ' Piiril ication  of 
/)- Aminophenol,"  U.S.  Patent  ;{.(i94.,')08,  1972. 

“f.  ,\.  Baron  and  R.  C.  Bennei',  "Pui'iliealion  of /)-Aminophenol," 
U.S.  Patent  3,717,880,  1973. 


269 


270  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


cleaner.  Only  APAP  would  accumulate:  all  other 
metabolites  are  naturally  occurring.  Even  micro- 
organisms could  be  collected  after  each  batch  and 
processed  into  a cake  for  use  as  a high  protein 
animal  feed. 

Biological  parameters 

MICROBIAL  PATHWAY 

A proposed  pathway  for  converting  aniline  to 
APAP  via  the  acetylation  of  an  intermediate,  p-amino- 
phenol,  is  shown  in  figure  l-A-1.  Various  tungi  have 
been  identified  in  which  these  reactions  occur. ^ 


^R.  V.  Smith  and  J.  P.  Rosazza,  "Microbial  Models  of  Mammalian 
Metabolism,"  J.  Pharmaceut.  Sci.  64:1737-1759,  1975. 

“R.  Smith  and  J.  P.  Rosazza,  "Microbial  Models  of  Mammalian 
Metabolism:  Aromatic  Hvdro.wlation,"  Arch.  Biochem.  Biophys. 
161:551-558,  1974. 

^V'.  R.  Munzner,  E.  Mutschler,  and  M.  Riimmel,  "Uberdie  mikro- 
biologiscbe  unwandlung  N-baltiger  substrate"  (Concerning  the 
Microbiological  Transformation  of  N-  containing  Substrate),  Plant 
Medica  15:97-103,  1967. 


Figure  l-A-1.  Bioconversion  of  Aniline  to  APAP^ 


NHCOCH3 


Aniline 


SaPAP  = N-acetyl-p-aminophenol  = acetaminophen  = p-acetamidophenol  = 
p-hydroxyacetanilide  = Tylenol  (trade  name  of  McNeil  Laboratories). 

SOURCE;  Genex  Corp. 


/Mternatively,  aniline  could  he  acetylated  directly 
forming  acetanilide,  which  in  turn  would  he  hydro.x- 
vlated  to  APAP.'“  A number  of  Streptomyces  spe- 
cies have  been  found  to  coiwert  acetanilide  to 
APAP. The  pathway  imoh  ing  p-aminophenol  was 
chosen  simply  because  the  conversion  efficiency  of 
acetic  acid  to  APAP  would  he  slightly  higher  if  acetic 
acid  entered  the  overall  reaction  at  the  second  step 
rather  than  at  the  first  step. 

HOST  MICRO-ORGANISMS 

The  most  suitable  micro-organism  for  jiroduction 
of  APAP  in  large-scale  fermentation  may  not  neces- 
sarilv  be  one  that  normally  metabolizes  aniline  or 
/i-aminophenol.  While  a h^ictei'ium  might  ser\e  as  a 
suitable  host  for  insertion  and  e.xpression  ol  the  req- 
uisite genes,  a yeast  may  represent  a better  choice.  It 
will  prohablv  more  closely  resemble  the  organism 
from  which  the  genes  are  isolated. 

Fermentation  efficiencies 

CONVERSION  EFFICIENCIES 

The  molar  and  weight  comersion  efficii’iicic's  for 
the  bioconversion  of  feedstock  to  product  are  pro- 
jected in  table  I-A-1.  I'be  biocom crsion  of  aniline  to 

'“Smith,  et  al.,  op.  oil. 

"Munzner,  el  al.,  op.  cil. 

'^R.  J.  Thei'iaull  and  I . It.  I.ongfield.  'Microbial  ( omeiMon  ot 
.Acetanilide  to  2 ’-Hvdro.wacetanilide  and  4’  Uydro.wacetanilide." 
Apt.  Microbiol.  15:1431-1436.  1967. 

‘"Ibid. 


Table  l-A-1. — Fermentation  Efficiencies  to  Meet  the 
Requirements  for  the  Production  of  Acetaminophen 
(APAP)  From  Aniline 

Overall  molar  conversion  efficiency  of: 


(a)  Aniline  to  APAP 90.25% 

(b)  Acetic  Acid  to  APAP 95.0 

Overaii  weight  conversion  efficiency  of: 

(a)  Aniline  to  APAPa 146.5 

(b)  Acetic  Acid  to  APAP® 239. 1 

Utiiization  of: 

(a)  Aniline  in  fermentation  broth 2.28lb/gal 

(b)  Acetic  acid  in  fermentation  broth ...  1 . 39/gal 

Production  of  APAP  in  broth 3.34  Ib/gal 

Batch  voiume 33,500  gal 

Recovery  efficiency 90.0  % 

Yield  of  APAP/batch 100,701  lb 

Number  of  batches/year 100 

Annual  yield  of  product 10,070,100  lb 


Overall  weight  conversion  of  precursor  to  APAP  ■ 

molecular  weight  of  APAP  y molarconvf  rston  effti  ? 

molecular  weight  of  precursor  of  precursor  to  APAP 

SOURCE:  Genex  Corp. 


Appendix  l-A—A  Case  Study  of  Acetaminophen  Production  • 271 


\1*  \l’  in\  ()l\  ('s  two  The  product  of  tlie  indi\  id- 

ual  reactions  for  each  step  represents  the  oxerall 
lom  ersion  efficiency.  ,\  molar  conversion  efficiency 
of  [)ercent  was  assumed  for  each  step.  This  value 
is  based  on  a multitude  of  reports  demonstrating 
similar  molar  conversion  efficiencies  for  analogous 
hiochemical  reactions  under  actual  fermentation 
conditions. 

PKOniCT  VIELI) 

I'he  yield  of  \l’  \l’  |)rojected  in  table  1 -.\-l  is  based 
on  estimating  a ratio  of  40  percent  w eight  to  v olume 
ti  e..  40  lb  ()er  100  gallons  (gal)  of  fermentation  broth) 
[trior  to  90  [tercent  recov  ery.  Such  a high  yield  is  per- 
mitted because  of  the  poor  solubility  of  .\P.AP  under 
o[)erating  conditions.  \s  a result,  high  levels  of  .\P.\P 
would  bav  e no  atlv  er.se  effect  on  the  host  micro-orga- 
nism. Use  of  insoluble  systems  in  fei  iiientation  has  in 
fact  been  reported  in  recent  years— e.g.,  in  certain 
microbial  transformations  of  steroids,  yields  of  40 
percent  may  result  due  to  the  insolubility  of  the 
product. 

“It  J Vhholl  immohill/cd  ( CIK.  " In  \niw;il  Reports  on  h'rrmcn- 
liilion  PriHvssrs,  \nl.  I I)  I’ci  lman  ((‘d.l  I.Vi-w  Vork:  Vcadriiiic 
Pi  rsN.  I!)77).  |)|) 


Economics 

PRODUCTION  REQUIREMENTS 

How  the  various  production  requirements  would 
be  met  during  the  microbial  transformation  of  ani- 
line to  APAP  is  summarized  in  tables  I-A-1  and  2.  Ani- 
line and  acetic  acid  would  not  be  added  to  the  fer- 
mentation broth  all  at  once  but  rather  step-wise  ac- 
cording  to  their  rates  of  conversion.  The  plant  would 
contain  two  50, 000-gal  fermenters,  which  in  the 
course  of  a year  would  yield  10  million  lb  of  APAP. 

PRODUCTION  COSTS 

The  costs  for  the  annual  production  are  summa- 
rized in  table  l-A-3.  I’hey  are  broken  down  into  their 
majoi'  components  and  are  expressed  both  as  annual 
costs  and  as  unit  costs.  Detailed  budgets  for  the  vari- 
ous cost  centers  are  shown  in  tables  I-A-4  through 
l-,\-10.  Materials  and  supplies  are  described  in  table 
I-.A-4;  labor  distribution  in  table  I-A-5;  utility  require- 
ments in  tables  l-A-6  through  I-A-8;  equipment  in 
table  l-A-9;  and  space  requirements  in  table  I-A-10. 
This  analysis  reveals  a unit  cost  of  APAP  equal  to 
$ 1.05/lb. 


Table  l-A-2.— Summary  of  Production  Conditions 
of  APAP 


Table  I-A-4.— Materials  and  Supplies  for 
Production  of  APAP 


Number  of  fermenters 
Size  of  fermenters ... 
Operating  volume  ... 

Cycle 

Batches  

^day  fermentation.  1-day  turn  around. 
SOURCE:  Cenex  Corp. 


Table  l-A-3.— Summary  of  Costs  of 
Production  of  APAP 


Annual  cost 

Cost/lb 

Materials  and  supplies  . . . . 

...  $ 6,133,802 

$0.6091 

Labor 

. ..  2,012,140 

0.1998 

Utilities 

630,200 

0.0626 

Equipment 

. ..  1,377,590 

0.1368 

Building 

439,399 

0.0436 

Total 

$10,593,131 

$1.05/lb 

Annual  production  = 10,070,1001b 

SOURCE:  Cenex  Corp. 


Materials 

Cost/batch 

Cost/year 

Fermentation 

Fishmeal  (1.5%  @ $0.155) 

$ 648.68 

$ 

64,868 

Glucose  (1.5%  @ $0.1535) 

Lard  oil  (2.5%  @ $0.325) 

Mineral  salts  (4,215  lb  @ $0.05074) 
Aniline  (76,250  lb  @ $0.42) 

642.40 

2,266.88 

213.77 

32,027.52 

64,240 

226,888 

21,377 

3,202,752 

Acetic  acid  (46,680  lb  @ $0,245)  . . 

11,436.60 

1 

,143,660 

Miscellaneous  (10%  of  basic 
materials) 

Subtotal 

377.17 

$47,613.02 

37,717 

$4,761,302 

Recovery 

Filter  aid  (0.2  Ib/gal  @ $13) 

$ 871.00 

$ 

87,100 

Other  chemicals  and  supplies. .. . 

1,600.00 

$ 

160,000 

Subtotal 

$ 2,471.00 

$ 

247,100 

Finishing 

Packaging  (1,255  bag  units 
at  $0.80) 

$ 1,004.00 

$ 

100,400 

Other  (labels,  stencils,  etc.) 

1,004.00 

$ 

100,400 

Subtotal 

$ 2,008.00 

$ 

200,800 

General  supplies 

Maintenance  (4%  of  capital  investment) 

$ 

425,900 

Other  (laboratory  office,  plant  miscellaneous)  . 
Total 

498,700 

$6,133,802 

SOURCE:  Cenex  Corp. 


2 

50,000  gal 
33,500  gal 
7a 
100 


272  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Table  l-A-5.— Labor  Distribution  for  Production  of  APAP 


Salary  and  wage  cost 


Category 

Man-hours  per  week 

Hourly  rate 

$/week 

$/year 

Supervision 

General  manager 

40 

20 

$ 800 

$ 41,600 

Superintendents 

80 

17 

1,360 

70,720 

Managers 

80 

15 

1,200 

62,400 

Supervisors 

320 

12 

3,840 

199,680 

Hourly  rated  employees,  services 
Laboratory 

Level  1 

80 

10 

800 

41,600 

Level  II 

80 

8 

640 

33,280 

Level  ill 

120 

6 

720 

37.440 

Level  IV 

40 

5 

200 

10,400 

Maintenance  and  engineering 

Level  1 

240 

10 

2,400 

124,800 

Level  II 

240 

8 

1,920 

99,840 

Level  III 

240 

6 

1,440 

74,880 

Level  IV 

160 

5 

800 

41,600 

Hourly  rated  employees,  production 

Fermentation  department 

Level  1 

200 

10 

2,000 

104,000 

Level  II 

240 

8 

1,920 

99,840 

Level  III 

80 

6 

480 

24,960 

Level  IV 

80 

5 

400 

20,800 

Recovery  department 

Level  1 

320 

10 

3,200 

166,400 

Level  II 

400 

8 

3,200 

166,400 

Level  III 

80 

6 

480 

24,960 

Level  IV 

120 

5 

600 

31,200 

Subtotal 

$1,476,800 

Add  overtime  @ 6%  x 1.5 

132,912 

Subtotal 

$1,609,712 

Add  fringe  benefits  @ 25% 

402,428 

Total  salaries  and  wages 


$2,012,140 


SOURCE:  Genex  Corp. 


Table  l-A-6.— Steam  Requirements  for 
Production  of  APAP 


Operation  Lb/batch 


Sterilization,  fermenters,  and  seed  tanks: 

Heating 52,100 

Holding 20,000 

Sterilization,  piping,  and  equipment  (other)  . . . 20,000 

Heating  acetaminophen  solution  (recovery) . . . 163,500 

Drying,  turbo  dryer 200,300 

General  purpose  usage 50,000 

Total 505,900 


Cost  at  S5.00/M  lb: 

Per  fermenter  batch  =$  2,530 
Per  year  (100  batches)  =$253,000 


SOURCE:  Genex  Corp. 


Appendix  l-A—A  Case  Study  of  Acetaminophen  Production  • 273 


Table  l-A-7.— Electricity  Requirements  for  Production  of  APAP 


Connected  load 

HP 

kW 

Units/batch 
(hours  operation) 

kWh 

Fermenters 

. . 200 

149 

144 

21,456 

Seed  tanks 

. . 47.5 

35 

24 

840 

Chillers 

. . 580 

433 

11 

4,763 

Air  compressor 

. . 275 

205 

86 

17,630 

Harvest  tank 

. . 100 

75 

11 

825 

Decanter  centrifuge 

. . 120 

90 

52 

4,680 

Process  tanks 

..300 

224 

19 

4,256 

Crystallizing  tanks 

..300 

224 

11 

2,464 

Turbo  dryer 

. . 30 

22 

23 

506 

Cooling  tower 

. . 40 

30 

144 

4,320 

Pumps  (est.  = 6 @ 7.5) 

. . 45 

34 

144 

4,896 

Lighting,  instruments  and  general  load 

Total  kWh 

@ 0.05/kWh  = $ 3,469  per  batch 

@ 100  batches/yr  = $346,900  per  year 

. . 25 

19 

144 

2,736 

69,372 

SOURCE:  Cenex  Corp. 


Table  l-A-8. — Water  Requirements  for 
Production  of  APAP 


Gal/batch 

Fermentation 35,000 

Tower  makeup 63,000 

Process  loss 100,000 

Chilled  water  makeup 30,000 

Direct  cooling 50,000 

General  use 25,000 

Total 303,000  gal 

Process  water  rate  =$1. 00/M  gals 
Cost  = $303/batch 

100  batches/yr  = $30,300/year 


SOURCE:  Cenex  Corp. 


274  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


Table  l-A-9.— Equipment  Costs  for 
Production  of  APAP 


$ 56,100 


585.000 

653.000 


132,000 

20,000 

48,000 


498.000 

650.000 

575.000 

210.000 

104,700 

24,000 

583,791 


583,790 

389,194 

$7,783,875 


$1,377,590 


SOURCE:  Cenex  Corp. 


Table  l-A-10.— Building  Requirements  for  Production  of  APAP 


Area 

Gross  space  ftVft^ 

Unit  values 

Cost 

Central  office 

940 

41.00b 

$ 38,540 

Laboratories 

4,500 

70.00b 

315,000 

Warehouse 

2,000/36,000 

27.00b 

54,000 

Batching 

1,000/30,000 

1.75 

52,500 

Fermentation  (including  seed)  . . . 

. . . 6,000/320,000 

1.75 

560,000 

Harvest,  filter 

. . . 3,500/169,000 

1.75 

295,750 

Processing,  crystallization 

. . . 8,700/470,000 

1.75 

822,500 

Drying,  finishing 

. . . 5,000/270,000 

1.75 

472,500 

Warehouse,  finished  product .... 

. ..  11,000/200,000 

27.00b 

297,000 

Auxiliary  equipment 

. ..  4,300/154,000 

1.75 

269,500 

Maintenance,  engineering 

. ..  11,500/207,000 

1.75 

362,250 

Total $3,539,540 


Amortization  over  30  years  @ 12%  compound  interest  $439,399<: 

®Unit  values  in  cubic  feet  except  where  noted  by  "b." 

“Unit  value  in  square  feet. 

“Amortization  = 0.12414  x total. 

SOURCE:  Cenex  Corp. 


Receiving  and  batching  area 
3 20,000  gal  steel  aniline  storage  tanks, 

insulated  and  cooled  - @ $47,000  $ 341,000 

2 20,000  gal  aluminum  acetic  acid  storage 

tanks,  insulated  and  cooled  - @ $71 ,300 342,600 

1 10,000  gal  steel  nitrogen  storage  tank  with 

controls  and  instruments 47,000 

1 10,000  gal  steel  lard  oil  storage  tank, 
insulated  and  heated 22,300 

1 10,000  gal  stainless  steel  Batch  tank  with 

programable  controller  and  agitator 59,500 

2 1,700  ft^  stainless  steel  Hopper  bins  with 

conveyors-®  $58,100  116,200 

1 Electric  forklift  truck 11,400 

Fermentation  and  seed  area 

1 150  gal  stainless  steel  seed  vessel,  fully 
instrumented 125,000 

1 2,500  gal  stainless  steel  seed  vessel,  fully 

instrumented 169,000 

2 50,000  gal  stainless  steel  fermenters,  fully 

instrumented  with  central  control  room  - 

@ $399,000  798,000 

Recovery  area 

1 50,000  gal  stainless  steel  process  tank, 

cooled,  agitated  and  insulated 195,000 

1 3,000  gal  steel  filter  aid  slurry  tank 
with  agitator 11,300 

1 Stainless  steel  continuous  decanter 

centrifuge 167,000 

2 100,000  gal  stainless  steel  process  tanks, 
insulated  with  external  steam  injection  heater, 

pump  and  agitator- @ $333,000  666,000 


1 20,000  gal  stainless  steel  side-entering  surge 
tank  with  agitator 

3 50,000  gal  stainless  steel  crystallizing  tanks, 

insulated  with  heavy  duty  cooling  coils  and 
top-mounted  agitator  - @ $195,000 

1 Stainless  steel  turbo  tray  dryer 

2 3,500  ft^  stainless  steel  hopper  bins  - 

@ $66,000  

1 Bagging  unit 

4 Stainless  steel  finished  product  conveyors  - 

@ $12,000  

Auxiliary  equipment 

3 1,500  c.f.m.  reciprocating  air  compressors  - 

@ $166,000  

Laboratory  and  office  equipment 

Chillers,  500  ton  total  capacity 

1 Cooling  tower,  1,500  g.p,m 

35  Pumps  and  motors,  various  sizes 

2 Dump  trucks  - $12,000  

Ventilation,  general  and  spot  - @ 7,5% 

of  equipment 

Piping,  general,  materials  and  installation  - 

@ 7.5%  of  equipment 

Miscellaneous  equipment  (hand  tools,  etc.)  - 

@ 5%  of  equipment 

Total 


Annual  charge  for  capital  recovery  over  10-year 
period,  with  12%  interest  compounded 
annually  ($7,783,875  x 0.17698) 


Appendix  I-B 

A Timetable  for  the  Commercial 
Production  of  Compounds  Using 
Genetically  Engineered  Micro- 
Organisms  in  Biotechnology 


ObJiH'tii’vs 

• I he  esiimalion  ut  the  |)ro|)oi  tions  of  \ ai  ioiis 
gi  oups  ot  eommei'C'ial  products  and  processes  tor 
\\  hich  recomhiiiant  l)\  \ (rI)N  \)  technolog\’  could 
he  a[)plicahle. 

• I lie  construction  of  timetables  to  indicate  [ilausi- 
hie  se(|ut'iices  of  commercial  de\elopments  that 
would  rt'sult  from  the  application  of  rDN'A  tech- 
nolof^N . 

Approaches 

The  follow  inj;  fi\  e industries  w ere  e\  aluated; 

1.  pharmaceutical. 

2.  agricultural. 

3.  food. 

4.  chemical,  and 
v"S.  energ\ . 

Ihe  manufacturing  processes  that  would  result 
from  the  application  of  rD\.A  technolog\'  would  be 
based  on  fermentation  technologx'.  Therefore,  a set 
of  parameters  w as  de\  eloped  to  ser\  e as  a guide  to 
assess  the  economics  of  applying  fermentation  tech- 
nolog\’  to  the  manufacture  of  products  currently 
manufactured  b\'  other  means. 

The  chemical  industry  generates  a large  number 
of  products  that  could  be  attributed  to  (and  is  in  this 
study)  the  other  four  industries  cited,  this  particular 
industry  was  focused  on  more  closely  than  the 
others.  The  following  factors  were  considered  in 
constructing  the  timetables  show  ing  the  applicability 
of  I'DN'.A  tecbnologx': 

• the  current  state  of  the  art  of  genetic  engineer- 
ing: 

• the  current  economic  limitations  of  fermenta- 
tion technology': 

• the  length  of  time  to  progress  from  a laboratory 
process  to  the  pilot  plant  to  large-scale  produc- 
tion: 

• the  plant  construction  time;  and 

• the  Go\  ernment  regulatory  agency  appro\  al  re- 


(|uired  (of  the  products  and  manufacturing 
processes,  not  of  the  rDNA  technology  per  se). 

Sources  of  information 

W'hile  much  of  the  information  compiled  for  this 
report  was  obtained  from  published  sources,  a con- 
siderable amount  came  from  prior  proprietary  stud- 
ies performed  by  (ienex  Corp.  In  the  latter  case,  in- 
formation is  used  that  is  not  proprietary,  although 
the  sources  must  remain  confidential.  In  this  connec- 
tion (iene.x  has  had  numerous  discussions  with  the 
technical  and  corporate  management  of  more  than 
100  large  companies  (generally  multibillion  dollar 
companies),  concei'ning  research  interests,  product 
lines,  and  market  trends.  Production  costs  are  ex- 
trapolated for  four  fermentation  plants  of  various 
sizes  and  capabilities.  (See  table  l-B-1 .) 

A group  of  Genex  scientists,  consisting  of  a bio- 
chemical engineer,  two  organic  chemists,  a biochem- 
ist, and  four  molecular  geneticists  rated  the  feasibili- 
ty of  devising  micro-organisms  to  produce  various 
chemicals  in  accordance  with  the  fermentation  con- 
ditions specified  in  table  l-B-1.  For  those  chemicals 
that  appeared  to  be  capable  of  being  produced  mi- 
crobiologically,  dates  were  assigned  for  the  times 
when  the  necessary  technology  would  be  achieved  in 
the  laboratory.  By  combining  both  technical  and  eco- 
nomic factors,  it  then  became  possible  to  project  a 
timetable  for  commercial  production.  (See  table 
l-B-2.) 

It  should  be  emphasized  that  an  extremely  con- 
servative approach  w'as  taken  in  considering  fermen- 
tation economics  over  the  next  10  years.  Only  the  rel- 
ativelv  poor  economics  of  conventional  batch  fer- 
mentation was  considered.  Immobilized  cell  proc- 
esses were  projected  to  be  15  years  away,  and  even 
then,  the  incremental  cost  savings  projected  (see 
table  I-B-1)  are  lower  than  the  incremental  cost  sav- 
ings currently  obtained  with  immobilized  cell  proc- 
esses. The  assumptions  made  here,  however,  did  in- 
clude reasonably  high  product  yields  and  highly  effi- 


275 


276  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


Table  Unit  Cost  Assumptions  for  the  Production  of  Chemicals  by 

Fermentation  After  Various  Intervals  of  Time 


Earliest  date 
(year) 

Size  of  plant 
(lb) 

Type  of 
fernnentation 

Product  yield 
(%) 

Annual  c^ 
excluding 
precursor 
($  millions) 

Unit  cost 
excluding 
precursor 
($/lb) 

Precursor 

Complete 
unit  cost 
($/lb) 

5 

50 

Ordinary  batch 

12 

23.5 

0.47 

Petrochemical^ 

0.66 

10 

100 

Ordinary  batch 

40 

24.5 

0.25 

Petrochemical 

0.44 

15 

200 

Immobilized  cells 

40 

25.5 

0.13 

Petrochemical 

0.32‘J 

20 

200 

Immobilized  cells 

40 

25.5 

0.13 

Carbohydrate‘S 

0.24d 

^Annual  costs  for  ordinary  batch  fermentation  were  estimated  from  proprietary  data.  Vaiues  obtained  for  the  immobiiized  ceii  exampies  are  computed  at  31.2  percent 
beiow  the  comparabie  vaiues  for  ordinary  batch  fermentation. 

^Average  cost  of  petrochemicai  equals  $0.17/lb.  At  90  percent  conversion  efficiency,  cost  contribution  of  petrochemical  equals  $0.19/lb  of  product. 

‘-Average  cost  of  carbohydrate  assumed  at  $0.04/lb  of  molasses  or  $0.02/lb  of  cellulose-containing  pellets  from  biomass  residue.  For  50  percent  free  sugar  content  of 
molasses,  cost  of  sugar  equals  $0.08/lb.  At  70  percent  conversion  efficiency  from  the  sugar,  cost  contribution  of  molasses  equals  $0.1 1/lb  of  product.  For  50  percent 
cellulose  content  in  the  biomass  pellets,  cost  of  cellulose  equals  $0.04/lb.  For  50  percent  conversion  efficiency  to  free  sugar,  followed  by  70  percent  conversion  effi- 
ciency from  the  sugar,  cost  contribution  of  the  pellets  also  equals  $0.1 1/lb  of  product. 

‘^These  unit  costs  may  be  further  reduced  to  $0.26  and  $0.17/lb.,  respectively,  for  products  whose  annual  U.S.  production  currently  exceeds  1 billion  lb.  Assumptions  In- 
clude reduction  in  precursor  cost  by  20  percent  (presumably  because  manufacturer  controls  supply  of  precursor);  reduction  in  unit  cost  of  immobilized  cell  process  by 
13  percent  (d)  and  42  percent  (e),  respectively;  maximum  of  80  percent  product  yield  (e);  and  a nearly  100  percent  bioconversion  efficiency  from  the  petrochemical 
precursor. 

SOURCE:  Genex  Corp. 


Table  1-6-2.— Basis  for  Estimating  the  Timetable  for 
Manufacture  of  Chemicals  by  Means  of  Microbial 
Processes 


Earliest  date 
for  commercial 
production^  is: 

If  all  the 
technology^ 
is  achieved 
by: 

And  if  bulk 
selling  prices's 
(in  1979 
dollars)  equal 
or  exceed: 

Assuming 
unit  costs'^  (in 
1979  dollars) 
equal  or 
exceed: 

5 years 

2 years 

$1. 32/lb 

$0.66/1  b 

10 

7 

0.88 

0.44 

15 

12 

0.64  (0.43) 

0.32  (0.26) 

20 

17 

0.48  (0.28) 

0.24  (0.17) 

®it  is  assumed  that  development  of  the  appropriate  manufacturing  facilities 
begins  at  least  5 years  prior  to  the  onset  of  producfion. 

technology  refers  to  both  genetic  and  biochemical  engineering.  Technology 
would  be  achieved  on  demonstrating  that  the  chemical  could  be  biologically 
produced  in  the  laboratory  at  commercially  desirable  yields  and  reaction  effi- 
ciencies. 

‘-It  is  assumed  that  all  bulk  selling  prices  are  marked  up  100  percent  from  the 
corresponding  unit  costs,  except  for  chemicals  whose  annual  U.S.  production 
currently  exceeds  1 billion  lb.  In  those  cases  the  bulk  selling  prices  (numbers 
in  parentheses)  are  assumed  to  be  marked  up  only  67  percent. 

‘^Unit  costs  were  obtained  from  table  l-B-1.  See  footnote  of  table  l-B-1  for  ex- 
planation of  numbers  in  parentheses. 

SOURCE:  Genex  Corp. 

cient  transformations  of  precursor  to  product,  but 
nothing  exceptional  with  respect  to  current  fermen- 
tation technology.  Indeed,  high  product  yields  and 
highly  efficient  reactions  would  he  expected  with 
genetically  engineered  micro-organisms. 

Two  points  should  he  stressed  that  place  these 
projections  on  the  low  side.  First,  they  exclude  cer- 
tain groups  of  products,  the  end  products  of  which 
could  not  be  microbially  processed,  although  their 
basic  constituents  could  be  produced  microbiologi- 
cally  (e.g.,  monomers  of  microbial  origin  could  form 
chemically  synthesized  polymers).  Second,  the  pro- 
jections exclude  naturally  occurring  products  of 


microbial  origin,  which  could  he  efleclixc  or  su|)('ri- 
or  substitutes  for  chemically  synlhf'sized  products 
that  could  not  he  manufactui  f'd  microhiologically.  .\s 
examples,  dyes  of  mici'ohial  origin,  such  as  pro- 
digiosin,  might  advantageously  rf'place  those  synthe- 
sized chemically,  hecausf'  their  toxicity  is  lower  than 
their  chemical  counterp;irts.  In  tlu*  case  of  plastics,  a 
new  generation  of  plastics  of  microbial  origin,  e,g., 
pullulans,  would  not  have  to  he  made  from  petro- 
chemical feedstocks  and  would  he  hiodegradahle. 

Explanation  of  tables 

Tables  l-H-;t  through  I-M-;J2  pix'sent  the  compounds 
from  two  points  of  \ i('w  , Tables  l-ll-.i  to  Ml- 1 0 grouj) 
the  compounds  by  industry  suhgrouped  h\  product 
catffgory.  TahU's  I-I5-It  to  l-lt-;i2  group  the  com- 
pounds by  product  category  ii'respi'ctis  e ot  industr\ 

The  tables  hasf'd  on  industry  present  end  use  d.ita 
for  each  compound:  e.g,,  in  the  pharmaceutical  in- 
dustry as|)irin  is  listf'd  as  an  aromatic  used  .is  an 
analgesic,  w hereas  in  the  chemical  industrv  .iniline  is 
listed  as  an  aromatic  used  as  a cyclic  intermedi.ite 
Thus,  tlu'  similarities  and  dillerf'iiees  between  com- 
pounds of  similar  origin,  i.e.,  product  e.itegory  are 
re\  ealed. 

Thff  tables  based  .solely  on  product  e.itegoiw  .ire 
dividf’d  into  two  tyjies:  one  type  |)ertaining  to  m.irket 
data  (tal)l('s  l-IMO,  I I.  and  ihi*  sul)s(‘(|U(*nt  odd  mim- 
herf'd  ones  through  table  l-H-.'l.'h,  and  the  other  jiei  ■ 
taining  to  technical  data  Ithe  e\en  niimhered  t.ihles 
from  l-H-12  through  I-I5-.52.) 

The  market  data  were  obtained  both  trom  piih- 
lishf'd  .sources  and  from  prior  pro|iriet.ir\  studies 


Appendix  l-B — A Timetable  for  the  Commercial  Production  of  Compounds  • 277 


Table  l-B-3.— Pharmaceuticals:  Small  Molecules 


Product  category 

End  use 

Amino  acids 

Phenylalanine 

. Intravenous  solutions 

Tryptophan 

. Intravenous  solutions 

Arginine 

. Therapeutic:  liver  disease 
and  hyperammonemia 

Cysteine 

. Therapeutic:  bronchitis  and 
nasal  catarrh 

Vitamins 

Vitamin  E 

. Intravenous  solutions, 
prophylactic 

Vitamin  Bu 

. Intravenous  solutions 

Aromatics 

Aspirin 

. Analgesic 

p-acetaminophenol 

Steroid  hormones 
Corticoids 

. Analgesic 

Cortisone 

. Therapeutic: 

anti-inflammatory  agent 

Prednisone 

. Therapeutic: 

anti-inflammatory  agent 

Prednisolone 

. Therapeutic: 

anti-inflammatory  agent 

Aldosterone 

Androgens 

. Therapeutic:  control  of 
electrolyte  imbalance 

Testosterone 

Estrogens 

. Therapeutic:  infertility, 
hypogonadism,  and 
hypopituitarism 

Estradiol 

Antibiotics 

. Prophylactic,  therapeutic: 
vaginitis 

Penicillins 

. Control  of  infectious  diseases 

Tetracyclines 

. Control  of  infectious  diseases 

Cephalosporins 

. Control  of  infectious  diseases 

Short  peptides 

Glycine-Histidine-Lysine.  . 

. Manufacturing  processes: 
tissue  culture 

SOURCE:  Genex  Corp. 


performed  by  (Jenex.  In  the  latter  case,  data  are  used 
that  are  not  proprietary  although  the  sources  must 
remain  confidential.  Market  \alues  were  estimated 
hy  multiplying  the  market  \olume  (total  amount  of 
product  sold  in  1978)  hy  the  hulk  cost  (unit  bulk  sell- 
ing price  in  1980).  Except  for  aromatics  and  ali- 
phatics.  all  market  data  represent  worldwide  esti- 
mates. Market  data  for  aromatics  and  aliphatics  are 
restricted  to  the  Ignited  States.  Data  that  could  not  be 
found  were  marked  not  a\  ailable  (N/A).  Compounds 
with  a high  market  \ alue  were  identified,  and  those 
that  could  he  produced  biologically  were  selected  for 
this  report. 

High  market  \ alues  were  relati\e  to  the  industry 
and  end  use  listing  of  each  compound.  For  example, 
with  respect  to  chemicals,  normally  only  cyclic  in- 


Table l-B-4.— Pharmaceuticals:  Large  Molecules 


Product  category 

End  use 

Peptide  hormones 

Insulin 

Endorphins 

Enkephalins 

ACTHa 

Glucagon  

Vasopressin 

Human  growth  hormone  . 

. . Control  of  diabetes 
. . Analgesics,  narcotics, 
prophylactics 
. . Analgesics,  narcotics, 
prophylactics 

. . Diagnostic:  adrenal  instability 
. . Therapeutic:  diabetes-induced 
hypoglycemia 
. . Therapeutic:  antidiuretic 
. . Therapeutic:  dwarfism 

Enzymes 

Glucose  oxidase 

Urokinase 

Asparaginase 

Tyrosine  hydroxylase  ... 

. . Diagnostic:  measurement  of 
blood  sugar 

. . Therapeutic:  antithrombotic 
. . Therapeutic:  antineopla§tic 
. . Therapeutic:  Parkinson’s 
disease 

Viral  antigens 

Hepatitis  viruses 

Influenza  viruses 

Herpes  viruses 

Varicella  virus 

Rubella  virus 

Reoviruses 

Epstein-Barr  virus 

. . Vaccine 
. . Vaccine 
. . Vaccine 
. . Vaccine 
. . Vaccine 

. . Vaccine:  common  cold 
. . Vaccine:  infectious 
mononucleosis, 
nasopharyngeal 
carcinoma,  Burkitt 
lymphoma 

Miscellaneous  proteins 

Interferon 

Human  serum  albumin  .. 
Monoclonal  antibodies  . . 

. . Control  of  infectious  diseases 
. . Therapeutic:  shock  and  burns 
. . Diagnostics:  hepatitis,  cancer, 
etc;  therapeutics 

Gene  preparations 

Sickle-cell  anemia 

Hemophilias 

Thallasemias 

. . Control  of  hereditary  disorder 
. . Control  of  hereditary  disorder 
. . Control  of  hereditary  disorder 

^Adrenocorticotropic  hormone. 
SOURCE:  Genex  Corp. 


termediates  with  production  volumes  (which  differ 
from  market  \ olumes)  exceeding  50  million  lb  were 
selected,  but  in  the  case  of  fla\  or  and  perfume  mate- 
rials, compounds  with  production  values  generally 
exceeding  1 million  lb  were  selected.  In  the  case  of 
many  pharmaceuticals,  clinical  importance'  was 
weighed  heavily  in  their  selection  process. 

The  technical  data  were  also  obtained  both  from 
published  and  proprietary  sources.  With  respect  to 
the  timetable  for  commercial  production,  the  stated 
length  of  time  is  the  time  required  to  develop  existing 
technology  (including  both  genetic  and  biochemical 
engineering)  to  the  point  where  it  may  be  applied  to 
appropriate  manufacturing  facilities  for  the  large- 
scale  production  of  the  desired  compounds.  These 
time  intervals  should  he  sufficient  for  undertaking 


278  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Table  l-B-5.— Food  Products 


Product  category 

End  use 

Amino  acids 
Glutamate 

Cysteine 

Aspartate 

. . Food  enrichment  agent, 
flavoring  agent 
. . Food  enrichment  agent, 
manufacturing  processes 
. . Flavoring  agent 

Vitamins 

Vitamin  C 

Vitamin  D 

. . Food  additive,  food 
enrichment  agent 
. . Food  enrichment  agent 

Aromatics 

Benzoic  acid 

. . Food  preservative 

Aliphatics 

Propionic  acid 

. . Food  preservative 

Short  peptides 

Aspartame 

. . Artificial  sweetener 

Enzymes 

Rennin 

Amyloglucosidase  . . . 
a-amylase 

Glucose  isomerase. . . 

Manufacturing  processes 
. . Manufacturing  processes 
. . Food  enrichment  agent, 
manufacturing  processes 
. . Manufacturing  processes: 
sweetener 

Nucleotides 

5’-IMPa 

5’-GMPb 

. . Flavoring  agent 
. . Flavoring  agent 

®5'-inosinic  acid. 
^5’-guanylic  acid. 
SOURCE;  Cenex  Corp. 


all  the  R&D  starting  from  the  current  knowledge 
base  necessary  to  demonstrate  that  the  desired  com- 
pounds can  he  biologically  produced  first  in  the 
laboratory  and  then  in  the  pilot  plant  at  commercial- 
ly desirable  yields  and  reaction  efficiencies.  The 
timetable  does  not  consider  delays  caused  by  con- 
struction of  new  facilities  nor  delays  required  to 
obtain  Ciovernment  regulatory  approval  of  new 
products. 

It  should  be  noted  that  in  the  technical  data  charts, 
when  glucose  is  listed  as  an  alternate  precursor  by 
fermentation,  other  carbohydrates,  e.g.,  cellulose 
and  cornstarch,  could  be  used.  Moreover,  if  glucose 
were  the  precursor  of  choice,  the  actual  feedstock 
would  probably  he  a commodity  like  molasses  as  op- 
posed to  pure  glucose. 

Summary 

Over  100  compounds  representing  17  different 
product  categories  that  span  the  five  industries 
under  evaluation  are  represented  in  table  l-B-10.  The 
current  market  value  of  all  these  products  exceeds 
$27  billion.  One  particular  compound,  methane,  ac- 
counts for  over  $12  billion.  The  even-numbered 


Table  l-B-G.— Agricultural  Products 


Product  category 

End  use 

Amino  acids 

Lysine 

Feed  additive 

Methionine 

Feed  additive 

Threonine 

Feed  additive 

Tryptophan 

Feed  additive 

Vitamins 

Nicotinic  acid 

Feed  additive 

Riboflavin  (B2) 

, Feed  additive 

Vitamin  C 

Feed  additive 

Aliphatics 

Sorbic  acid 

Feed  preservative 

Antibiotics 

Penicillins 

, Feed  additive,  prophylactic 

Erythromycins 

. Feed  additive,  prophylactic 

Peptide  hormones 

Bovine  growth  hormone 

, Growth  promoter 

Porcine  growth  hormone 

. Growth  promoter 

Ovine  growth  hormone 

, Growth  promoter 

Viral  antigens 

Foot-and-mouth  disease  virus 

. Vaccine 

Rous  sarcoma  virus 

. Vaccine 

Avian  leukemia  virus 

. Vaccine 

Avian  myeloblastosis  virus. . . 

. Vaccine 

Enzymes 

Papain 

. Feed  additive 

Glucose  oxidase 

. Feed  preservative 

Pesticides 

Microbial 

. Insecticide 

Aromatic 

. Insecticide 

Inorganics 

Ammonia 

. Fertilizer 

SOURCE:  Genex  Corp. 


tables  from  l-B-12  lo  I-B-;12  |)rojecI  that  within  20 
years  all  th(^s(!  |)ioducts  could  Ix'  manulactured 
using  geiK'tically  engineered  microbial  strains  on  a 
more  econotnical  basis  than  using  today  s coinen- 
tional  technologi(?s.  In  many  cas(‘s,  the  time  i ('(|uired 
to  apply  genetically  engineered  strains  in  eommerci.il 
fermentations  could  he  reduced  loas  little  .is  ,'i  \ e.irs 
The  impact  of  geiK'tic  engineering  on  selected 
markets  is  shown  in  table  l-M-.kt.  Only  five  |)roduct 
categories  are  (xmsiden’d  here,  and  in  one.  .imino 
acids,  only  a lew  of  the  compounds  com|irising  it  .ire 
evaluated.  The  products  represented  in  the  In  e i .ite 
gories  currently  ha\  (>  a total  market  \ aloe  exc  eeding 
$800  million,  llowexcr,  within  20  \ears  this  m.ii  ki't 
value  could  rise  to  oxer  $.">  billion  tin  1!)80  doll.irsi 
due  largely  to  the  application  of  gi-netic  engineering 
In  a number  of  cases,  the  desired  products  would 
most  likely  not  he  av  ailable  in  signilic  .ml  <|u.mlilies  it 
not  for  the  application  of  genetic  engineering  lech 
nology. 


Appendix  l-B— A Timetable  for  the  Commercial  Production  of  Compounds  • 279 


Table  Chemicals:  Aliphatics 


Table  l-B-9. — Energy  Products 


Compound 

Acetic  acid^ 

Acrylic  acid® 

Adipic  acid^ 

Bis  (2-ethylhexyl)  adipate 

Citronellal 

Citronellol 

Ethanol^ 

Ethanolamine 

Ethylene  glycol® 

Ethylene  oxide® 

Geraniol 

Glycerol® 

Isobutylene 

Itaconicacid 

Linalool 

Linalyl  acetate 

Methane 

Nerol 

Pentaerythritol 

Propylene  glycol® 

Sorbitol 

a-terpineol 

a-terpinyl  acetate 


End  use 

Miscellaneous  acyclic 
Miscellaneous  acyclic 
Miscellaneous  acyclic 
Plasticizer 

Flavor/perfume  material 
Flavor/perfume  material 
Miscellaneous  acylic 
Miscellaneous  acyclic 
Miscellaneous  acyclic 
Miscellaneous  acyclic 
Flavor/perfume  material 
Miscellaneous  acyclic 
Miscellaneous  acyclic, 
flavor/perfume  material 
Plastics/resin 
Flavor/perfume  material 
Flavor/perfume  material 
Primary  petroleum  product 
Flavor/perfume  material 
Miscellaneous  acyclic 
Miscellaneous  acylic 
Miscellaneous  acyclic 
Flavor/perfume  material 
Flavor/perfume  material 


®lndicates  compounds  also  identilied  by  the  Massachusetts  Institute  of 
Technology.  The  following  additional  chemicals  were  identified  by  MIT  as 
amenable  to  biotechnological  production  methods:  acetaldehyde,  acetoin, 
acetone,  acetylene,  acrylic  acid,  butadiene,  butanol,  butyl  acetate, 
butyraldehyde.  dihydroxyacetone.  ethyl  acetate,  ethyl  acrylate,  ethylene,  for- 
maldehyde. isoprene.  isopropanol,  methanol,  methyl  ethyl  ketone,  methyl 
acrylate,  propylene,  propylene  oxide,  styrene,  vinyl  acetate. 

SOURCE:  Cenex  Corp.  and  the  Massachusetts  Institute  of  Technology. 


Table  l-B-8.— Chemicals:  Aromatics  and 
Miscellaneous 


Product  category 

End  use 

Aromatics 

Aniline 

Benzoic  acid 

Cresols 

Phenol 

Phthalic  anhydride 

Cinnamaldehyde 

Diisodecyl  phthalate 

Dioctyle  phthalate 

. . Cyclic  intermediate 
. . Cyclic  intermediate 
. . Cyclic  intermediate 
. . Cyclic  intermediate 
. . Cyclic  intermediate 
. . Flavor/perfume  material 
. . Plasticizer 
. . Plasticizer 

Inorganics 

Ammonia 

Hydrogen  

. . Manufacturing  processes 
. . Manufacturing  processes 

Enzymes 

Pepsin 

Bacillus  protease 

. . Manufacturing  processes 
. . Manufacturing  processes 

Mineral  leaching 
Transition  metals  (cobalt, 
nickel,  manganese,  iron).  . . . 

Inorganic  intermediates; 
catalysts 

Biodegradation 

. . Removal  of  organic 
phosphates,  aryl 
sulfonates,  and 
haloaromatics 

Product  category 

End  use 

Enzymes 

Ethanol  dehydrogenase. 
Hydrogenase 

Manufacturing  processes 
Manufacturing  processes 

Biodegradation 

Petroleum  byproducts  removal 

Aliphatics 

Methane 

Ethanol  

Fuel 

Fuel 

Inorganics 

Hydrogen 

Fuel 

Mineral  leaching 
Uranium 

Fuel 

SOURCE:  Genx  Corp. 


Table  l-B-10.— Total  Market  Values  for  the 
Various  Product  Categories 


Product  category 

Number  of 
compounds 

Current  value 
($  millions) 

Amino  acids 

9 

$ 1,703.0 

Vitamins 

6 

667.7 

Enzymes 

11 

217.7 

Steroid  hormones 

6 

376.8 

Peptide  hormones 

9 

263.7 

Viral  antigens 

9 

N/A 

Short  peptides 

2 

4.4 

Nucleotides 

2 

72.0 

Miscellaneous  proteins  . . . 

2® 

300.0 

Antibiotics 

4b 

4,240.0 

Gene  preparations 

3 

N/A 

Pesticides 

2b 

100.0 

Aliphatics: 

Methane 

1 

12,572.0 

Other 

24= 

2,737.5 

Aromatics 

10= 

1,250.9 

Inorganics 

2 

2,681.0 

Mineral  leaching 

5 

N/A 

Biodegradation 

N/A 

N/A 

Totals 

107 

$27,186.7b 

®Only  two  of  a number  of  compounds  are  considered  here. 

*h"hese  numbers  refer  to  major  classes  of  compounds;  not  actual  numbers  of 
compounds. 

'-These  numbers  refer  only  to  those  compounds  representing  the  largest 
market  volume  in  classes  specified  in  the  text. 

'^Current  value  excluding  methane  = $14,614,700,000. 

SOURCE:  Cenex  Corp. 


SOURCE:  Cenex  Corp. 


280  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animais 


Table  l-B-11.— Amino  Acids:  Market  Information 


Current  market  data 


Market 

volume  Bulk  cost  Market  value 
Compound  1,0001b  $/lb  ($  millions) 

Arginine 900  12.73  11.46 

Aspartate 3,000  2.86  8.6 

Cysteine 600  22.75  13.6 

Glutamate 600,000  1.80  1,080.0 

Lysine 129,000  2.10  258.0 

Methionine 210,000  1.40  294.0 

Phenylalanine....  300  38.18  11.46 

Threonine 300  58.18  16.2 

Tryptophan 225  43.18  9.71 


SOURCE:  Compiled  by  Genex  Corp.  from  data  in  references  1,2,  and  3. 


Table  l-B-12.— Amino  Acids:  Technical  Information 


Compound 

Typical  synthetic 
process 

Typical  precursor 

Is  precursor 
renewable/non- 
renewable 
limited 

Alternate 
precursor  by 
fermentation 

Time  to  implement 
commercial  fermentation  by 
genetically  engineered 
strain 

Arginine 

fermentation 

glucose 
and  NH4^ 

renewable 

— 

5 yrs. 

Aspartate 

fermentation 

fumaric  acid 
and  ammonia 

limited 

— 

5yrs. 

Cysteine 

extraction 

protein 

hydrolysis 

renewable 

— 

5 yrs. 

Glutamate 

fermentation 

glucose 
and  NH/ 

renewable 

— 

5 yrs. 

Lysine 

fermentation 

glucose 
and  NH/ 

renewable 

— 

5 yrs. 

Methionine 

chemical 

/3-methylmercapto 

propionaldehyde 

nonrenewable 

glucose 
and  NH4^ 

10  yrs. 

Phenylalanine. . . 

chemical 

a-acetamino- 
cinnamic  acid 

limited 

glucose 
and  NH."’ 

5 yrs. 
5 yrs. 

Threonine 

fermentation 

glucose 
and  NH/ 

renewable 

— 

5 yrs. 

Tryptophan 

chemical 

a-ketoglutaric 

phenylhydrazone 

nonrenewable 

glucose 
and  NH/” 

5 yrs. 

^Ammonium  ion. 


SOURCE:  Compiled  by  Genex  Corp.  from  data  in  references  2,  3, 4,  and  5. 

Table  l-B-13.— Vitamins:  Market  Information 


Current  market  data 

Market 

volume  Bulk  cost  Market  value 

Compound 1,000  lb $/lb ($  millions) 

Nicotinic  acid.  ..  . 1,400  1.82  2.5 

Riboflavin  (Bj).  ..  . 22  15.40  0.34 

Vitamin  Bi2  22  6,991.60  153.8 

Vitamin  C 90,000  4.50  405.0 

Vitamin  D 12  42.50  0.51 

Vitamin  E 3,641  29.00  105.6 


SOURCE:  Compiled  by  Genex  Corp.  from  data  in  references  1,6,7, 8.  and  9. 


Appendix  l-B—A  Timetable  for  the  Commercial  Production  of  Compounds  • 281 


Table  l•B-14.— Vitamins:  Technical  Information 

Compound 

Typical  synthetic 
process 

Typical  precursor 

Is  precursor 
renewable/non- 
renewable 
limited 

Alternate 
precursor  by 
fermentation 

Time  to  implement 
commercial  fermentation  by 
genetically  engineered 
strain 

Nicotinic  Acid  . . 

chemical 

alkyl  a-subst. 

nonrenewable 

glucose 
and  NH/® 

lOyrs. 

Riboflavin  (Bi).  . . 

fermentation 

pyridines 

glucose 

renewable 

— 

lOyrs. 

Vitamin  Bu 

fermentation 

carbohydrates 

renewable 

— 

lOyrs. 

Vitamin  C 

semisynthetic 

glucose  or 
sorbitol 

renewable 

— 

10  yrs. 

Vitamin  D 

fermentation 

glucose 

renewable 

glucose 

10  yrs. 

Vitamin  E 

extraction 

wheat  germ  oil 

limited 

glucose 

15  yrs. 

^Ammonium  ion. 

SOURCE:  Compiled  by  Gene*  Corp.  from  data  In  references  4,  7.  8, 10, 11,  and  12. 


Table  l-B-15.— Enzymes:  Market  Information 


Current  market  data 


Compound 

Market 

volume 

1,0001b 

Bulk  cost 
$/lb 

Market  value 
($  millions) 

a-amylase 

600 

19.33 

11.6 

Amyloglucosidase 

600 

.00 

12.0 

Asparaginase .... 

(Information  not  available) 

Bacillus  protease. 

1,000 

8.28 

8.2 

Ethanol 

dehydrogenase  . 

(Information  not  available) 

Glucose  isomerase 

100 

400  00 

40  0 

Glucose  oxidase  . 

5 

160.00 

0.80 

Hydrogenase  .... 

(Information  not  available) 

Papain 

200 

59.00 

11.8 

Pepsin 

10 

380.00 

3.8 

Rennin 

24 

696.00 

40.0 

Tyrosine 

hydroxylase  .... 

(Information  not  available) 

Urokinase 

60;900IUa 

89.5 

^lU  = international  units. 

SOURCE:  Compiled  by  Gene*  Corp.  from  data  in  references  9, 13, 14, 15,  and  16. 


282  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Table  l-B-16.— Enzymes:  Technical  Information 

Is  precursor 

Time  to  implement 

renewable/non- 

Alternate 

commercial  fermentation  by 

Typical  synthetic 

renewable 

precursor  by 

genetically  engineered 

Compound 

process 

Typical  precursor 

limited 

fermentation 

strain 

a-amylase 

fermentation 

molasses 

renewable 

— 

5 yrs. 

Amyloglucosidase 

fermentation 

molasses 

renewable 

— 

5 yrs. 

Asparaginase .... 

extraction 

tissue  culture 

renewable 

glucose 
and  NH/ 

5 yrs. 

Bacillus  protease. 
Ethanol 

fermentation 

molasses 

renewable 

— 

5 yrs. 

dehydrogenase 

(Information  not  available) 

glucose 
and  NH."" 

10  yrs. 

Glucose 

isomerase 

fermentation 

glucose 
and  NH/3 

renewable 

5 yrs. 

Glucose  oxidase  . 

fermentation 

molasses 

renewable 

— 

5 yrs. 

Hydrogenase  .... 

(Information  not  available) 

glucose 
and  NH4" 

10  yrs. 

Papain 

extraction 

papaya 

renewable 

glucose 
and  NH-^ 

5 yrs. 

Pepsin 

fermentation 

molasses 

renewable 

— 

5 yrs. 

Rennin 

fermentation 

molasses 

renewable 

— 

5 yrs. 

Tyrosine 

extraction 

tissue  culture 

renewable 

glucose 
and  NH/ 

5 yrs. 

Urokinase 

extraction 

tissue  culture 

renewable 

glucose 

5 yrs. 

^Ammonium  ion. 

SOURCE:  Compiled  by  Cenex  Corp.  from  data  in  references  4,  5, 13, 14. 16, 17,  and  18. 


Table  l-B-1 7.— Steroid  Hormones:  Market 
Information 


Current  market  data 

Market 

volume  Bulk  cost  Market  value 
Compound 1,000  lb $/lb ($  millions) 

Corticoids 305.8 

Cortisone N/A  208.84  N/A 

Prednisone N/A  467.62  N/A 

Prenisolone N/A  463.08  N/A 

Aldosterone N/A  N/A  N/A 

Androgens  10.8 

Testosterone  ....  (Information  not  available) 

Estrogens 60.2 

Estradiol (Information  not  available) 


SOURCE:  Compiled  by  Cenex  Corp.  from  data  in  references  1 and  4. 


Appendix  l-B— A Timetable  for  the  Commercial  Production  of  Compounds  • 283 


Table  1 

B-18.— Steroid  Hormones;  Technical  Information 

Compound 

Typical  synthetic 
process 

Typical  precursor 

Is  precursor 
renewable/non- 
renewable 
limited 

Alternate 
precursor  by 
fermentation 

Time  to  implement 
commercial  fermentation  by 
genetically  engineered 
strain 

Corticoids 

Cortisone 

Prednisone .... 

Predisolone 

Aldosterone 

semisynthetic 

diosgenin  or 
stigmasterol 

renewable 

glucose 

10  yrs. 

Androgens 

Testosterone  . . 

semisynthetic 

chemical 
modification 
of  cholesterol 

renewable 

glucose 

10  yrs. 

Estrogens 

Estradiol 

semisynthetic 

chemical 
modification  of 
cholesterol 

renewable 

glucose 

10  yrs. 

SOURCE;  Compiled  by  Cenex  Corp.  (rom  data  in  references  4, 19, 20,  21,  and  22. 


Table  l-B-19.— Peptide  Hormones;  Market 
Information 


Current  market  data 

Market 

volume  Bulk  cost  Market  value 
Compound  1,0001b  $/lb  ($  millions) 

ACTHa n7a  5^6 

Bovine  growth 

hormone 0.0  0.0  0.0 

Endorphins (Information  not  available) 

Enkephalins (Information  not  available) 

Glucagon (Information  not  available) 

Human  growth 

hormone N/A  N/A  75.0 

Insulin N/A  N/A  183.1 

Ovine  growth 

hormone 0.0  0.0  0.0 

Porcine  growth 

hormone 0.0  0.0  0.0 

Vasopressin (Information  not  available) 


aAdrenocorticotropic  hormone. 

SOURCE:  Compiled  by  Cenex  Corp.  from  data  in  reference  4. 


284  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


Table  l-B-20.— Peptide  Hormones;  Technical  Information 


Compound 

Typical  synthetic 
process 

Typical  precursor 

Is  precursor 
renewable/non- 
renewable 
limited 

Alternate 
precursor  by 
fermentation 

Time  to  implement 
commercial  fermentation  by 
genetically  engineered 
strain 

ACTHa 

Bovine  growth 

extraction 

adrenal  cortex 

limited 

glucose 

and 

5yrs. 

hormone 

extraction 

anterior  pituitary 

limited 

glucose 
and  NH**" 

5yrs. 

Endorphins 

extraction 

brain 

limited 

glucose 
and  NH4^ 

5yrs. 

Enkephalins  . . . . 

extraction 

brain 

limited 

glucose 
and  NH4" 

5yrs. 

Glucagon  

Human  growth 

extraction 

pancreas 

limited 

glucose 
and  NH/ 

5yrs. 

hormone 

extraction 

anterior  pituitary 

limited 

glucose 
and  NH4" 

5 yrs. 

Insulin 

Ovine  growth 

extraction 

pancreas 

limited 

glucose 
and  NH/ 

5yrs. 

hormone 

Porcine  growth 

extraction 

anterior  pituitary 

limited 

glucose 
and  NH/ 

10  yrs. 

hormone 

extraction 

anterior  pituitary 

limited 

glucose 
and  NH4*^ 

10  yrs. 

Vasopressin  . . . . 

extraction 

posterior  pituitary 

limited 

glucose 
and  NH/ 

5 yrs. 

^Adrenocorticotropic  hormone, 

^Ammonium  ion. 

SOURCE:  Compiled  by  Cenex  Corp.  from  data  in  references  4,  23,  and  24. 


Table  l•B-21.— Viral  Antigens:  Market  Information 


Current  market  data 

Market 

volume  Bulk  cost  Market  value 

Compound 1,000  lb $/lb ($  millions) 

Avian  leukemia 

virus (Information  not  available) 

Avian  myeloblastosis 

virus (Information  not  available) 

Epstein-Barr  virus  0.0  0.0  0.0 

Hepatitis  virus  ..  . 0.0  0.0  0.0 

Herpesvirus 0.0  0.0  0.0 

Hoof  and  mouth 

disease  virus  ..  0.0  0.0  0.0 

Influenza  virus  . . . (Information  not  available) 

Reoviruses 0.0  0.0  0.0 

Rous  sarcoma 

virus (Information  not  available) 

Rubella  virus (Information  not  available) 

Varicella  virus.  . . . (Information  not  available) 


SOURCE:  Compiled  by  Cenex  Corp.  from  data  in  reference  4, 


Appendix  l-B— A Timetable  for  the  Commercial  Production  of  Compounds  • 285 


Table  l-B-22.— Viral  Antigens:  Technical  Information 

Compound 

Typical  synthetic 
process 

Typical  precursor 

Is  precursor 
renewable/non- 
reneywable 
limited 

Alternate 
precursor  by 
fermentation 

Time  to  implement 
commercial  fermentation  by 
genetically  engineered 
strain 

Avian  leukemia.  . . 

virus 

Avian 

myeloblastosis. 

virus 

(Information  not  available) 
(Information  not  available) 

glucose 
and  NH/® 

glucose 
and  NHx"" 

5yrs. 

5yrs. 

Epstein-Barr  virus 

tissue  culture 

lymphoblasts 

renewable 

glucose 
and  NH/ 

5 yrs. 

Hepatitis 

viruses 

(Information  not  available) 

glucose 
and  NHx"^ 

5 yrs. 

Herpes 

viruses 

(Information  not  available) 

glucose 
and  NH4+ 

5 yrs. 

Hoof  and  mouth  . . 
disease  virus 

(Information  not  available) 

glucose 
and  NHx^ 

5 yrs. 

Influenza 

viruses 

(Information  not  available) 

glucose 
and  NHx^ 

10  yrs. 

Reoviruses 

(Information  not  available) 

glucose 
and  NH/ 

15  yrs. 

Rous  sarcoma  . . . 
virus 

(Information  not  available) 

glucose 
and  NH4+ 

5 yrs. 

Rubella 

virus 

tissue  culture 

duck  embryonic 
cells 

renewable 

glucose 
and  NH4+ 

5 yrs. 

Varicella 

virus 

(Information  not  available) 

glucose 

NH4-" 

5 yrs. 

^Ammonium  ion. 

SOURCE:  Compiled  by  Cenex  Corp,  from  data  in  references  4 and  25. 


Table  l-B-23.— Short  Peptides,  Nucleotides,  and 
Miscellaneous  Proteins:  Market  Information 


Current  market  data 

Market 

volume  Bulk  cost  Market  value 
Product  category  1,0001b  $/lb  ($  millions) 

Short  peptides^ 

Aspartame 40  110.00  4.4 

Glycine-histidine- 

lysine (Information  not  available) 

Nucleotides^’ 

5’-IMPc 4,000  12.00  48.0 

5'-GMPd 2,000  12.00  24.0 

Miscellaneous  proteins^ 

Interferon N/A  N/A  50.0 

Human  serum 

albumin 250  1,000.00  250.0 

Monoclonal 

antibodies (Information  not  available) 


^Data  from  references  4 and  26. 
j ^Data  from  references  4 and  27. 

•'  '-5'-inosinic  acid. 

'^5’-guanylic  acid. 

®Data  from  reference  4. 

SOURCE:  Compiled  by  Cenex  Corp. 

'] 

|i 


286  • Impacts  of  Applied  Genetics— Micro-Organisms,  Piants,  and  Animais 


Table  l-B-24.— Short  Peptides,  Nucleotides,  and  Miscellaneous  Proteins:  Technical  Information 


Is  precursor 

Time  to  implement 

renewable/non- 

Alternate 

commercial  fermentation  by 

Typical  synthetic 

renewable 

precursor  by 

genetically  engineered 

Product  category 

process 

Typical  precursor 

limited 

fermentation 

strain 

Short  peptides^ 

Aspartame 

chemical 

phenylalanine  & 

renewable 

glucose 

5 yrs. 

aspartic  acid 

and 

Glycine-histidine- 
lysine 

extraction 

human  serum 

renewable 

glucose 
and  NH/ 

5yrs. 

Nucleotides’^ 

5’-IMPd 

extraction 

yeast 

renewable 

glucose 

and 

10  yrs. 

5’-GMPe 

extraction 

yeast 

renewable 

glucose 
and  NH/ 

10  yrs. 

Miscellaneous  proteins* 

Interferon 

extraction  or 

leukocytes, 

renewable 

glucose 

5 yrs. 

tissue  culture 

lymphoblasts, 
or  fibroblasts 

and  NH4+ 

Human  serum 
albumin 

extraction 

human  serum 

renewable 

glucose 
and  NH/ 

5 yrs. 

Monoclonal 

antibodies 

somatic  cell 

various  cells 

renewable 

glucose 

10  yrs. 

hybridization 

and  NH/ 

®Data  from  references  4 and  27, 
^Ammonium  ion. 

‘"Data  from  references  4 and  28. 
'^S'-inosinic  acid. 

®5'-guanyiic  acid. 

*Data  from  reference  4. 

SOURCE:  Complied  by  Genex  Corp. 


Table  l-B-25.— Antibiotics,  Gene  Preparations,  and 
Pesticides:  Market  Information 


Current  market  data 

Product  category 

Market 

volume 

1,0001b 

Bulk  cost 
$/lb 

Market  value 
($  millions) 

Antibiotics^ 
Peniciilins 

49,300 

22.11 

1,080.0 

Tetracyclines 

29,300 

34.13 

1,000.0 

Cephalosporins  . . 

4,210 

114.00 

480.0 

Erythromycins  . . . 

(Information  not  available) 

Gene  preparations*> 

Sickle  cell  anemia 

0.0 

0.0 

0.0 

Hemophilias 

0.0 

0.0 

0.0 

Thallasemias  .... 

0.0 

0.0 

0.0 

Pesticides’^ 
Microbiai 

N/A 

N/A 

25.0 

Aromatics 

N/A 

N/A 

75.0 

®Data  from  references  4,  28,  and  9. 
^Data  from  references  4 and  29. 
'"Data  from  references  4 and  30. 
SOURCE:  Compiled  by  Genex  Corp. 


Appendix  l-B — A Timetable  for  the  Commercial  Production  of  Compounds  • 287 


Table  l-B-26.— Antibiotics,  Gene  Preparations,  and  Pesticides:  Technical  Information 


Is  precursor 

Time  to  implement 

renewable/non- 

Alternate 

commercial  fermentation  by 

Typical  synthetic 

renewable 

precursor  by 

genetically  engineered 

Product  category 

process 

Typical  precursor 

limited 

fermentation 

strain 

Antibiotics^ 
Penicillins 

fermentation 

lactose  & 

limited 

10  yrs. 

semisynthetic 

nitrogenous  oils 

Tetracyclines  .... 

fermentation 

lactose  & 

limited 

— 

lOyrs. 

nitrogenous  oils 

Cephalosporins  . . 

fermentation 

lactose  & 

limited 

— 

10  yrs. 

nitrogenous  oils 

Erythromycins  . . . 

fermentation 

lactose  & 

limited 

— 

10  yrs. 

nitrogenous  oils 

Gene  preprations‘> 
Sickle  cell  anemia 

(No  process  exists 

glucose 

15  yrs. 

currently) 

and  NH/'^ 

Hemophilias 

(No  process  exists 

glucose 

20  yrs. 

currently) 

and  NH4^ 

Thallasemias  .... 

(No  process  exists 

glucose 

20  yrs. 

currently) 

and  NH/ 

Pesticides’^ 
Microbial 

fermentation 

molasses  & 

renewable 

5 yrs. 

fishmeal 

Aromatics 

semisynthetic 

naphthalene 

nonrenewable 

— 

10  yrs. 

^Data  from  references  4.  5,  28,  31.  and  32.  •'Data  from  references  4 and  30. 

^Data  from  reference  4.  •'Ammonium  ion. 

SOURCE.  Compiled  by  Cenex  Corp. 


Table  I B-27.— Aliphatics:  Market  Information 


Current  market  data 

Compound 

Market 

volume  Bulk  cost  Market  value 

1,0001b  $/lb  ($  millions) 

Acetic  acid 

823,274 

0.23 

189.4 

Acrylic  acid 

46,503 

0.43 

20.0 

Adipic  acid 

181,097 

0.50 

90.5 

Bis  (2-ethylehexyl) 

43,015 

0.49 

21.1 

adipate 

Citronellal 

394 

3.90 

1.5 

Citronellol 

1,443 

4.50 

6.5 

Ethanol 

1,048,000 

0.24 

251.5 

Ethanolamine.  . . . 

320,236 

0.46 

147.3 

Ethyleneglycol  .. 

3,137,000 

0.31 

972.5 

Ethylene  oxide  . . . 

525,113 

0.36 

189.0 

Geraniol 

2,307 

3.25 

7.5 

Glycerol 

116,612 

0.54 

63.0 

Isobutylene 

597,712 

0.95 

567.2 

Itaconic  acid 

200 

0.83 

0.2 

Linalool 

3,341 

2.60 

8.7 

Linalyl  acetate  . . . 

1,535 

3.50 

5.4 

Methane 

878,000,000 

0.013 

11,573.0 

Nerol 

462 

4.20 

1.9 

Pentaerythritol . . . 

117,085 

0.62 

72.6 

Propionic  acid  . . . 

62,848 

0.21 

13.2 

Propylene  glycol  . 

525,527 

0.73 

173.4 

Sorbic  acid 

20,000 

2.15 

43.0 

Sorbitol 

160,267 

0.36 

57.7 

a-terpineol 

2,416 

1.28 

3.0 

a-terpinyl  acetate. 

1,066 

1.30 

1.4 

SOURCE:  Compiled  by  Cenex  Corp.  from  data  in  references  1 , 4,  9,  and  33. 


288  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Table 

-B-28.— Aliphatics:  Technical  Information 

Compound 

Typical  synthetic 
process 

Typical  precursor 

Is  precursor 
renewable/non- 
renewable 
limited 

Alternate 
precursor  by 
fermentation^ 

Time  to  implement 
commercial  fermentation  by 
genetically  engineered 
strain^* 

Acetic  acid 

chemical 

methanol 
or  ethanol 

nonrenewable 

glucose 

10  yrs. 

Acrylic  acid 

chemical 

ethylene 

nonrenewable 

glucose 

lOyrs. 

Adipic  acid 

chemical 

phenol 

nonrenewable 

glucose 

10  yrs. 

Bis(2-ethylhexyl). . 
adipate 

chemical 

phenol 

nonrenewable 

glucose 

20  yrs. 

Citronellal 

chemical 

isobutylene 

nonrenewable 

glucose 

20  yrs. 

Citronellol 

chemical 

isobutylene 

nonrenewable 

glucose 

20  yrs. 

Ethanol 

chemical 

ethylene 

nonrenewable 

glucose 

5 yrs. 

Ethanolamine 

chemical 

ethylene 

nonrenewable 

glucose 

10  yrs. 

Ethylene  glycol  . . . . 

chemical 

ethylene 

nonrenewable 

glucose 

5 yrs. 

Ethylene  oxide 

chemical 

ethylene 

nonrenewable 

glucose 

5 yrs. 

Geraniol 

chemical 

isobutylene 

nonrenewable 

glucose 

20  yrs. 

Glycerol 

chemical 

soap  manuf. 

nonrenewable 

glucose 

5 yrs. 

Isobutylene 

chemical 

petroleum 

nonrenewable 

glucose 

10  yrs. 

Itaconicacid 

fermentation 

molasses 

renewable 

.... 

5 yrs. 

Linalool 

chemical 

isobutylene 

nonrenewable 

glucose 

20  yrs. 

Linalyl  acetate 

chemical 

isobutylene 

nonrenewable 

glucose 

20  yrs. 

Methane 

chemical 

natural  gas 

nonrenewable 

sewage 

10  yrs. 

Nerol 

chemical 

isobutylene 

nonrenewable 

glucose 

20  yrs. 

Pentaerythritol  . . . . 

chemical 

acetaldehyde  & 
formaldehyde 

nonrenewable 

glucose 

10  yrs. 

Propionic  acid 

chemical 

ethanol  & 
carbon  monoxide 

limited 

glucose 

10  yrs. 

Propylene  glycol  . . 

chemical 

propylene 

nonrenewable 

glucose 

10  yrs. 

Sorbic  acid 

chemical 

crotonaldehyde  & 
malonic  acid 

nonrenewable 

glucose 

15yrs. 

Sorbitol 

chemical 

glucose 

renewable 

.... 

10  yrs. 

Qf-terpineol 

chemical 

isobutylene 

nonrenewable 

glucose 

20  yrs. 

a-terpinyl  acetate  . 

chemical 

isobutylene 

nonrenewable 

glucose 

20  yrs. 

^Wherever  glucose  is  mentioned,  other  carbohydrates  may  be  substituted,  including  starch  and  cellulose. 

^In  many  cases  these  times  are  based  on  more  readily  developed  fermentations  using  nonrenewable  or  limited  hydrocarbons  as  precursors. 
SOURCE:  Compiied  by  Genex  Corp.  from  data  in  references  4,  33,  34,  and  35. 


Table  l-B-29.— Aromatics:  Market  Information 


Current  market  data 

Compound 

Market 

volume 

1,0001b 

Bulk  cost 
$/lb 

Market  value 
($  millions) 

Aniline 

187,767 

0.42 

78.9 

Aspirin 

32,247 

1.41 

45.5 

Benzoic  acid 

36,822 

0.47 

17.3 

Cinnamaldehyde . . . . 

1,098 

2.10 

3.4 

Cresols 

94,932 

0.54 

51.2 

Diisodecyl 
phthalate 

151,319 

0.42 

63.6 

Dioctyl  phthalate. . . . 

391,131 

0.42 

164.3 

p-acetaminophenol . . 

20,000 

2.65 

53.0 

Phenol  

1,431,000 

0.36 

515.2 

Phthalic  anhydride  . . 

646,289 

0.40 

258.5 

SOURCE:  Compiled  by  Cenex  Corp.  from  data  in  references  1 and  9. 


Appendix  l-B— A Timetable  for  the  Commercial  Production  of  Compounds  • 289 


Table  l-B-30.— Aromatics:  Technical  Information 


Compound 

Typical  synthetic 
process 

Typical  precursor 

Is  precursor 
renewable/non- 
renewable 
limited 

Alternate 
precursor  by 
fermentation 

Time  to  implement 
commercial  fermentation  by 
genetically  engineered 
strain 

Aniline 

chemical 

benzene 

nonrenewable 

aromatic® 

10  yrs. 

Aspirin 

chemical 

phenol 

nonrenewable 

aromatic 

5 yrs. 

Benzoic  acid 

chemical 

tar  oil 

nonrenewable 

aromatic 

10  yrs. 

Cinnamaldehyde . . . 

chemical 

benzaldehyde 

acetaldehyde 

nonrenewable 

aromatic 

20  yrs. 

Cresols 

chemical 

phthalic 

anhydride 

nonrenewable 

aromatic 

10  yrs. 

Diisodecyl 

chemical 

coal  tar 

nonrenewable 

aromatic 

20  yrs. 

phthalate 

Dioctyl 

chemical 

coal  tar 

nonrenewable 

aromatic 

20  yrs. 

phthalate 

p-acetaminophenol . 

chemical 

nitrobenzene 

nonrenewable 

aromatic 

5 yrs. 

Phenol  

chemical 

coal  tar 

nonrenewable 

aromatic 

10  yrs. 

Phthalic 

chemical 

coal  tar 

nonrenewable 

aromatic 

15  yrs. 

anhydride 

^Aromatic  refers  to  benzene  or  benzene  derivative.  Eventually  it  Is  anticipated  that  lignin,  a renewable  resource,  would  serve  as  a precursor. 
SOURCE:  Compiled  by  Genex  Corp.  from  data  In  references  4 and  35. 


Table  l-B-31.— Inorganics  and  Mineral  Leaching: 
Market  Information 


Current  market  data 

Market 

volume  Bulk  cost  Market  value 

Product  category  1,0001b $/lb ($  millions) 

Inorganics 

Ammonia 33,400,000  0.06  2,004.0 

Hydrogen 451,000  0.15  677.0 

Mineral  leaching 

Uranium (Information  not  available) 

Transition  metals.  (Information  not  available) 


(cobalt,  nickel,  manganese,  iron) 


SOURCE:  Compiled  by  Genex  Corp.  from  data  in  reference  4. 


Table  l-B-32.— Inorganics  and  Mineral  Leaching:  Technical  Information 


Is  precursor  Time  to  implement 

renewable/non-  Alternate  commercial  fermentation  by 
Typical  synthetic  renewable  precursor  by  genetically  engineered 


Product  category process Typical  precursor limited fermentation strain 

Inorganics 

Ammonia chemical  waterandcoke  nonrenewable  nitrogen(air)  15yrs. 

Hydrogen catalytic  petroleum  nonrenewable  waterandair  15yrs. 

reforming 

Mineral  leaching 

Uranium (Information  not  available) 

Transition  metals.  (Information  not  available) 


(cobalt,  nickel,  manganese,  iron) 


SOURCE:  Compiled  by  Genex  Corp.  from  data  in  references  4 and  35. 


290  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


Table  l-B-33.— Projected  Growth  of  Selected  Markets 
Involving  Applications  of  Genetic  Engineering 


Product  category 

Current  market 
$ millions 

Projected 
market 
in  20yrs. 
$ millions 

Amino  acids® 

300 

900 

Miscellaneous  proteins . 

300 

1,000 

Gene  preparations 

0 

too 

Sfiort  peptides 

5 

2,100 

Peptide  hormones 

260 

1,000 

Totals 

865 

5,100 

®Only  four  amino  acids  are  considered  here. 
SOURCE:  Genex  Corp. 


References 

1.  Chemical  Marketing  Reporter,  March  and  April 
1980. 

2.  Hirose,  Y.,  and  Okada,  H.,  “Microbial  Production 
of  Amino  Acids,”  in  Microbial  Technology:  Micro- 
bial Processes,  vol.  1,  H.  J.  Peppier  and  D.  Perlman 
(eds.)  (New  York:  Academic  Press,  1979),  pp. 
211-240. 

3.  Hirose,  Y.,  Sano,  K.,  and  Shibai,  H.,  "Amino 
Acids,”  in  Annual  Reports  on  Fermentation  Proc- 
esses, vol.  2,  G.  T.  Tsao  and  D.  Perlman  (eds.) 
(New  York:  Academic  Press,  1978),  pp.  155-190. 

4.  Genex  Corp.  proprietary  information. 

5.  Weinstein,  L.,  “Chemotherapy  of  Microbial  Dis- 
eases," in  The  Pharmacological  Basis  of  Therapeu- 
tics, 5th  ed.,  L.  S.  Goodman  and  A.  Gilman  (eds.) 
(New  York:  MacMillan  Publishing  Co.,  Inc.,  1975), 
pp.  1,090-1,247. 

6.  Cohn,  V'.  H.,  “Fat-Soluble  Vitamins:  Vitamin  K and 
Vitamin  E,”  in  The  Pharmacological  Basis  of  Ther- 
apeutics, 5th  ed.,  L.  S.  Goodman  and  A.  Gilman 
(eds.)  (New  York:  MacMillan  Publishing  Co.,  Inc., 
1975),  pp.  1,591-1,600. 

7.  Florent,  J.,  and  Nitiet,  L.,  “Vitamin  Bij,”  in  Micro- 
bial Technology:  Microbial  Processes,  vol.  1,  H.  J. 
Peppier  and  D.  Perlman  (eds.)  (New  York:  Aca- 
demic Press,  1979),  pp.  497-520. 

8.  Perlman,  D.,  “Microbial  Process  for  Riboflavin 
Production,”  in  Microbial  Technology:  Microbial 
Processes,  vol.  1,  H.  J.  Peppier  (eds.)  (New  York: 
Academic  Press,  1979),  pp.  521-528. 

9.  Synthetic  Organic  Chemicals,  March  and  April 
1980. 

10.  Burns,  J.  J.,  "Water  Soluble  Vitamins,  The  Vdta- 
min  B Complex,”  in  The  Pharmacological  Basis  of 
Therapeutics,  5th  ed.,  L.  S.  Goodman  and  A. 
Gilman  (eds.)  (New  York:  MacMillan  Publishing 
Co.,  Inc.,  1975),  pp.  1,549-1,563. 

11.  Greengard,  P.,  “Water  Soluble  Vitamins,  The  Vita- 


min B Complex,”  in  The  Pharmacological  Basis  of 
Therapeutics,  5th  ed.,  L.  S.  Goodman  and 
Gilman  (eds.)  (New  York:  MacMillan  Publishing 
Co.,  Inc.,  1975),  pp.  1,549-1,563. 

12.  Straw,  J.  A.,  "Fat-Soluble  V itamins:  Vitamin  D,"  in 
The  Pharmacological  Basis  of  Therapeutics,  5th 
ed.,  L.  S.  Goodman  and  A.  Gilman  (eds.)  (New 
York:  MacMillan  Publishing  Co.,  Inc.,  1975),  pp. 
1,579-1,590. 

13.  Aunstrup,  K.,  “Industrial  Approach  to  Enzyme 
Production,”  in  Biotechnological  Af)plications  of 
Proteins  and  Enzymes,  Z.  Bohak  and  N.  Sharon 
(eds.)  (New  York:  Academic  Press,  1977),  pp. 
39-50. 

14.  Aunstrup,  K.,  Andresen,  ().,  Falch,  E.  Z.,  and  Miel- 
sen,  T.  K.,  “Production  of  Microbial  Enzymes,"  in 
Microbial  Technology:  Microbial  Processes,  \ol.  1, 
H.  J.  Peppier  and  D.  Perlman  (eds.)  (New  Vork: 
Academic  Press,  1979),  pj).  282-31  1. 

15.  Bernard  Wolnak  &.  Associates,  in  Food  Prod.,  July 
1978,  p.  41. 

16.  Solomons,  G.  L.,  “The  Microbial  Production  of  En- 
zymes,” in  Biotechnological  A/yplications  of  Pro- 
teins and  Enzymes,  Z.  Bohak  and  N.  Sharon  (eds.) 
(New  York:  Academic  Press,  1977),  pp.  51-61. 

17.  Chemical  and  Engineering  News,  .Apr.  14,  1980,  p. 
6. 

18.  Levine,  W.  G.,  “Anticoagulants,  .Antithrombotic, 
and  Thrombolytic  Drugs,"  in  I'he  Pharmacological 
Basis  of  Therapeutics,  5th  ed.,  E.  S.  (loodman  and 
A.  Gilman  (:eds.)  (New  York:  MacMillan  Publish- 
ing Co.,  Inc.,  1975),  p.  1,365. 

19.  Gilman,  A.  (i.,  and  Muriad,  F.,  “.Androgt'ns  and 
Anabolic  Steroids,"  in  I'he  Pharmacological  Basis 
of  Therapeutics,  5th  (h1.,  E.  S.  (lOodman  and  A 
Gilman  (eds.)  (New  Voi'k:  MacMillan  Publishing 
Co.,  Inc.,  1975),  |)p.  1,451-1,471. 

20.  Gilman  A.  and  Mui'iad,  F.,  "l■,strogens  and  Pro- 
gestins,”  in  I'he  Pharmacological  Basis  of  Ihrr- 
apeutics,  5th  ed.,  E.  S.  Goodman  and  A Gilman 
(eds.)  (New  York:  MacMillan  Publishing  ( o.,  Inc  . 
1975),  pp.  1,423-1,450. 

21.  Haynes,  R.  C.,  and  Earner,  J.,  “ Adrenocoi  lico- 
tropic  Hormone:  .Adrenocorticol  Steroid.s  and 
Their  Synthetic  .Analogs:  Inhibitors  of  Vdreno- 
corticol  Steroid  Bio.synthesis,"  in  I he  Pharmaco- 
logical Basis  of  I'herapeutics,  5th  ed  , 1 S (lood- 
man  and  ,A.  (iilman  (eds.)  (N(’\\  Vork:  MacMillan 
Publishing  Co.,  Inc.,  1975),  [)p.  1.4721. 506. 

22.  Sebek,  ().  K.,  and  Perlman,  1)..  Alierohi.il  leans 
formation  of  Steroid.s  and  Sterols,"  in  Microbial 
Technology:  Microbial  Procc.sscs,  \of  I II  .1  Pi’p- 
pier  and  1).  Perlman  (eds.)  (.\ew  V ork  Vc  ademii 
Press,  1979),  pp.  483-496. 

23.  Brazeau  P.,  ".Agents  Alleeling  the  Renal  ( onsci 


Appendix  t-B— A Timetable  for  the  Commercial  Production  of  Compounds  • 291 


\ ation  ot  \\  aler,"  in  The  Pharmacolog,ical  Basis  of 
Therapeutics,  5th  eel.,  L.  S.  (ioodman  and  A. 
(iilman  (eds.)  (New  ^o^k:  MacMillan  Publishing 
Inc..  1975),  pp.  858-859. 

24.  Haynes,  R.  C..  and  Larnei',  J.,  "Insulin  and  Oral 
Hypoglycemic  Drugs:  (ilucagon."  in  The  Pharma- 
cological Basis  of  I'herapeutics,  5th  ed.,  L.  S.  Good- 
man and  A.  Gilman  teds.)  (.New  \ork:  Mac.Millan 
Publishing  Go  . Inc.,  1975).  pp.  1,507-1,533. 

I 25.  Jawetz,  K.,  Melnick,  J.  1,.,  and  Adelherg,  K.  Z.,  Re- 
views of  Medical  Microhiolog}’,  11th  ed.,  (Los 
-Altos,  GaliL:  Lange  .Medical  Publications,  1974). 

j 26.  I'haler.  M.  M..  Biochem.  Biophys.  Bes.  Comm. 

\ 54:562.  1973. 

' 27.  Nakao.  N ..  "Microbial  Produclion  of  Nucleosides 
and  Nucleotides,"  in  .Microbial  Technology:  Micro- 
bial Processes,  \ol.  1,  H.  J.  Peppier  and  D.  Perlman 
(eds.)  (New  \ork:  Academic  Press,  1979),  pp. 
312-355. 

28.  Perlman,  D..  Microbial  Production  of  -Antibiot- 
ics," in  .Microbial  Technoloey:  Microbial  Processes, 
\()l.  1 H.  J.  Peppier  and  1).  Perlman  (eds.)  (New 
\ ork:  Academic  Press,  1979),  pp.  241-281. 

29.  Weatherall.  D.  J.,  and  C legg.  J.  B.,  "Recent  De\el- 
opments  in  the  Molecular  (ienetics  of  Human 
Hemoglobin,"  (,'e// 16:467-480,  1979. 


30.  Ignoffo,  C;.  M.,  and  Anderson,  R.  F.,  "Bioinsec- 
ticides,” in  Microbial  Technology:  Microbial  Proc- 
esses, vol.  1,  H.  J.  Peppier  and  D.  Perlman  (eds.) 
(New  York:  -Academic  Press,  1979),  pp.  211-240. 

31.  Gorman,  M.,  and  Huber,  F.  M.,  “jS-Lactam  Anti- 
biotics,” in  Annual  Reports  on  Fermentation  Proc- 
esses, \ol.  2,  G.  T.  Tsao  and  D.  Perlman  (eds.) 
(New  ^'ork:  -Academic  Press,  1978),  pp.  203-222. 

32.  Sbibata,  M.,  and  liyeda,  M.,  "Microbial  Transfor- 
mation of  Antibiotics,”  in  Annual  Reports  on 
Fermentation  Processes,  vol.  2,  G.  T.  Tsao  and  D. 
Perlman  (eds.)  (New  York:  Academic  Press,  1978), 
p|).  267-304. 

33.  Lockwood,  L.  B.,  "Production  of  Organic  Acids  by 
Fermentation,”  in  Microbial  Technology:  Microbial 
Processes,  \ol.  1,  H.  J.  Peppier  and  D.  Perlman 
(eds.)  (New  \'ork:  Academic  Press,  1979),  pp. 
367-372. 

34.  Roberts,  J.  D.,  and  Caserio,  M.  C.,  Basic  Principles 
of  Organic  Chemistry  (New  York:  Benjamin,  Inc., 
1964),  p.  1,096. 

35.  W'indholz,  M.  (ed.).  The  Merck  Inde^,  9th  ed.  (Rah- 
way, N.J.:  Merck  &.  C'o.,  1976). 


Appendix  I-C 

Chemical  and  Biological  Processes 


A comparison  was  made  of  waste  stream  pollution 
for  cliemical  and  biological  processes.  Ideally,  the 
comparison  should  he  between  the  two  processes 
used  in  the  production  of  the  same  end  product. 
Since  such  data  do  not  currently  exist  at  the  in- 
dustrial level,  the  comparison  was  made  between  the 
chemical  production  of  a mixture  of  chemicals  and 
the  biological  production  of  alcohol  and  antibiotics. 
One  noteworthy  parameter  is  the  5-day  biochemical 
oxygen  demand  (BODS)  —the  oxygen  required  over  a 
5-day  period  by  organisms  that  consume  degradable 
organics  in  the  waste  stream.  If  the  oxygen  demand 
is  too  high,  the  discharge  of  such  a stream  into  a 
body  of  water  will  deplete  the  dissolved  oxygen  to 
the  point  that  it  threatens  aquatic  life.  An  important 
variable  that  must  be  considered  along  with  the  BOD 
is  the  COD  (the  chemical  oxygen  demand).  Large  dif- 
ferences between  the  COD  and  BOD  of  a waste  sys- 
tem can  indicate  the  presence  of  nonhiodegradahle 
substances.  Although  the  conventional  process 
stream  shown  in  table  I-C-1  has  less  BOD5  than  the 
biological  process  stream,  its  COD  content  probably 
means  that  nonbiodegradables  are  present,  and 
specialized  waste  treatment  is  necessary. 

BOD  is  one  area  where  traditional  fermentation 
based  processes  have  posed  pollution  problems. 
Batch  fermentation  processes  typically  generate 
large  quantities  of  dead  cells  and  residual  nutrients 
that  cause  a large  BOD  if  they  are  dumped  directly 
into  a dynamic  aquatic  environment.  (See  table  I-C-1.) 
This  difficulty  can  be  circumvented  by  the  use  of 
spent  cell  material  as  an  animal  feed  supplement  or 


Table  I-C-1.— Waste  Stream  Pollution  Parameters: 
Current  Processes  v.  Biological  Processes 

Compounds:  IVlixed  chemicals,  including  ethylene 
oxide,  propylene  oxide,  glycols,  amines,  and  ethers 


Pollution  parameters 

Current 

processes 

Biological 

processes 

Alkalinity  (mg/I) 

4,060 

0 

BOD53(mg/l) 

1,950 

4,000-12,000 

Chlorides  (mg/I) 

430-800 

0 

CODb(mg/l) 

7,970-8540 

5,000-13,000 

Oils  (mg/I) 

547 

0 

pH 

9.4-9.8 

4-7 

Sulfates  (mg/I) 

655 

0 

Total  nitrogen  (mg/I) 

1,160-1,253 

50-200 

Phosphates  (mg/I) 

0 

50-200 

®5-day  biological  oxygen  demand. 

*^Chemical  oxygen  demand. 

SOURCE:  Office  of  Technology  Assessment. 


as  a fertilizer.  These  a|)|)lications  ha\(>  Imumi  inten- 
sively in\  estigated  and  ha\  (t  met  u ilh  success. 

Because  of  the  renewed  inlc'resl  gnK'ralefl  hy  the 
potentials  of  genetic  ('iigineering,  souk*  traditional 
fermentation  .systems  ar«f  Ix'ing  reih'signed.  Immo- 
hilization  allows  the  rmise  of  c(*lls  that  would  other- 
wise he  discarded.  These  .systems  c;m  he  used  con- 
tinuously for  se\eral  months  as  compared  with  the 
usual  fermentation  tim(>  in  a hatch  process  of  about 
one  week  or  less.  Immohilizc'd  operations  create 
waste  cells  much  U'ss  often  than  h;itch  s\stems.  .md 
therefore  generate  less  BOI ). 


292 


Appendix  I-D 

The  Impact  of  Genetics  on  Ethanol — 

A Case  Study 


Objective 

This  study  examines  how  genetics  can  and  will  af- 
fect the  utilization  of  biomass  for  liquid  fuels  produc- 
tion. There  are  two  major  areas  where  genetics  are 
applicable.  One  is  in  plant  breeding  to  impro\  e avail- 
ability (both  quantity  and  quality)  of  biomass  re- 
sources (with  existing  and  pre\  iously  unused  land); 
the  second  is  in  the  application  of  both  classical 
mutation  and  selection  procedures  and  the  new  ge- 
netic engineering  techniques  to  develop  more  effi- 
cient microbial  strains  for  biomass  conversion.  Ex- 
amples of  goals  in  a plant  breeding  program  would 
include  impro\  ements  in  pbotosynthetic  efficiencies, 
increased  carbohydrate  content,  decreased  or  modi- 
fied lignin  content,  adaptation  of  high  productixity 


plants  to  poor  quality  land,  improved  disease  resist- 
ance, and  so  forth.  However,  the  focus  here  is  entire- 
ly on  the  second  area,  the  use  of  genetics  to  improve 
microbial-based  conversion  to  produce  ethanol. 

In  order  to  assess  the  type  and  extent  of  im- 
provements in  micro-organisms  that  might  benefit 
ethanol  production,  its  process  technology  and 
economics  must  first  be  examined.  An  overview  of 
the  biomass  conversion  technology  is  presented  in 
figure  I-D-1;  processes  are  defined  mainly  on  the 
basis  of  the  primary  raw  material  and  the  type  of 
pretreatment  required  to  produce  mono-  or  di- 
saccharides prior  to  fermentation.  In  addition,  there 
are  several  alternative  fermentation  routes  to  pro- 
duce ethanol;  these  are  characterized  by  the  type  of 
micro-organisms  and  will  be  examined  with  the  in- 


Figure  I-D-1. —An  Overview  of  Alternative  Routes  for  Conversion  of  Biomass  to  Ethanol 


Primary  raw 
material 

Sugar 

(cane  or  beet) 

Starch 
(corn,  wheat 
or  tuber 
crop) 

Cellulosic  biomass 
(agricultural  or 
forest  residue) 

Pretreatment 

Extraction 

Gelatiniza- 

tion 

Grinding,  possible 
delignification 

Sucrose 

inversion 

Liquefac- 
tion, saccha- 
rifi  cation 

Acid  or  enzymatic 
hydrolysis 

Fermentable 

substrate 

Glucose/ 

fructose 

Glucose/ 

maltose 

Glucose/cellobiose 

xylose/xylobiose 

Fermentation  of 
sugar  to  ethanol 

Yeast 

Zymomonas 

Anaerobic  bacteria 

Product 

recovery 

Ethanol  and 

for  fuel 

Residue 
for  feed 

The  arrows  designate  the  fermentation  substrate  used  by  each  type  of  microorganism. 

SOURCE:  Massachusetts  Institute  ofTechnology. 


293 


294  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


tent  of  quantifying  the  potential  impact  of  genetic  im- 
provement on  each  one.  It  is  interesting  to  note  that 
each  type  of  organism  has  its  substrate  restrictions, 
and  only  the  anaerobic  bacteria  such  as  Clostridium 
thermosaccharolyticum  and  C.  thermohydrosulfori- 
cum  can  utilize  all  of  the  available  substrate. 

Substrate  pretreatment 

Pretreatment  refers  to  the  processing  that  is  re- 
quired to  convert  a raw  material  such  as  sugarcane, 
starch,  or  cellulosic  biomass  to  a product  that  is 
fermentable  to  ethanol.  In  most  cases,  the  pretreat- 
ment is  either  extraction  of  a sugar  or  hydrolysis  of  a 
polysaccharide  to  yield  a mono-  or  disaccharide. 

EXTRACTION  OF  SUGAR 

Sugar  crops  such  as  sugarcane,  sugar  beets,  or 
sweet  sorghum  are  highly  desirable  raw  materials 
for  producing  ethanol.  These  crops  contain  high 
amounts  of  sugars  as  sucrose.  In  addition,  the  yield 
of  fermentable  material  per  acre  is  high;  sugarcane 
and  sugar  beets  yield  7.5  and  4.1  dry  tons  of  biomass 
per  acre,  respectively.’ 

Sugar  is  extracted  from  cane  or  beets  with  hot 
water  and  then  recrystallized.  The  resulting  sugars 
are  utilized  directly  by  organisms  having  invertase 
activity  (to  split  sucrose  to  glucose  plus  fructose). 
Molasses,  a sugary  byproduct  of  the  crystallization  of 
sucrose,  may  also  contain  sucrose  although  in  most 
cases  it  is  inverted  with  acid. 

The  primary  use  for  sugar  crops  is  food  sugar. 
Sugar  sells  for  over  20  cents/lb.  Molasses,  which  cur- 
rently sells  for  about  $100/ton  (about  10  cents/lb 
sugar)  is  used  extensively  as  an  animal  feed.  Substan- 
tial amounts  of  both  sugar  and  molasses  are  im- 
ported into  the  United  States  for  food  uses  and  are 
therefore  unavailable  for  ethanol  production.  There 
are  proposals  to  increase  sugar  production  for  use  as 
an  energy  crop;  however,  this  will  require  the 
development  of  new  land  for  sugar  production. 

STARCH 

The  primary  raw  material  for  ethanol  fermenta- 
tion in  the  United  States  is  cornstarch.  Corn  proc- 
essed by  wet  milling,  yields  about  36  lb  of  starch 
from  each  56  lb  bu;  this  amount  of  starch  will  pro- 
duce 2.5  gal  of  absolute  ethanol.  Corn  yields  are 
typically  80  to  120  bu/acre  so  that  200  to  300  gal  of 
ethanol  can  be  derived  per  acre  of  corn  per  year. 

Pretreatment  of  starch  is  initiated  by  a gelatiniza- 
tion  step  whereby  a starch  slurry  is  heated  for  5 min 
at  105°  C.  After  cooling  to  98°  C,  a-amylase  is  added 


'Paul  B.  Weisz  and  John  F.  Marshall,  Science  206:24.  1979. 


to  break  down  the  starch  to  about  15DE  (dextrose 
equivalents).  This  process  of  liquefaction  reduces  the 
viscosity  such  that  the  solution  can  be  easily  mixed. 
After  further  cooling  to  30°  C,  glucoamviase  is  added 
along  with  a starting  culture  of  yeast  so  that  saccha- 
rification and  fermentation  proceed  simultaneousiv. 
The  resulting  fermentation,  to  produce  typically  8 to 
10  percent  ethanol  (v'olume  per  volume),  requires  42 
to  48  hr  for  completion.  This  compares  with  a 16-  to 
20-hr  fermentation  if  sugar  as  molasses  or  cane  juice 
is  used  as  the  substrate.  Thus,  the  use  of  starch  re- 
quires the  addition  of  enzymes  prior  to  and  during 
fermentation,  as  well  as  large  fermenter  capacity  as  a 
consequence  of  the  slower  fermentation  time  com- 
pared with  sugar  substrates. 

Improvement  in  the  economy  of  ethanol  fermenta- 
tion based  on  starch  is  possible  by  developing  a 
micro-organism  that  can  produce  a-amylase  and 
glucoamylase  and  thus  eliminate  the  need  to  add 
these  enzymes.  Since  the  rate  of  fermentation  de- 
pends on  the  rate  of  starch  hydrolysis,  increased  lev  - 
els of  glucoamylase  may  enhance  the  rate  of  starch 
hydrolysis  and  thus  increase  the  rate  of  ethanol  [)ro- 
duction.  This  would  lower  the  capital  re(iuirements 
as  well  as  the  cost  of  enzyme  addition. 

CELLULOSIC  BIOMASS 

Processes  for  the  utilization  of  cellulosic  biomass 
to  produce  liquid  fuels  all  have  three  features  in  com- 
mon; 

1.  They  employ  some  means  of  |)retreatment  to  at 
least  effect  some  initial  size  rc'duction  and.  moi'c 
often,  cause  a disassociation  of  lignin  and  cellu- 
lose; 

2.  they  involve  either  acid  or  enzymatic  hydrolysis 
of  the  cellulose  and  hemicellulose  to  |)roduce 
mono-  and  disaccharides;  and 

3.  they  employ  fermentation  to  |)roduce  (Mhanol  or 
some  other  chemical. 

A wide  variety  of  process  schemes  have  been  pro- 
posed for  the  conversion  of  cellulosic  biomass  to 
liquid  fuels;  a summary  of  the  major  steps  in  two 
acid  hydrolysis  and  three  enzymatic  hydrolysis 
schemes  in  shown  in  figures  I-I)-2  and  l-l)-3.  The  iti- 
itial  size  reduction  is  re{]uired  to  increase  the 
amount  of  biomass  surface  area  that  can  he  con- 
tacted with  acid,  solvent,  steam,  enzymes,  or 
chemicals  that  might  he  used  to  di.sassociate  the 
cellulose  and  hemicellulose  from  the  lignm 
Pretreatments  that  have  been  investigated  to 
facilitate  the  process  are  summarized  in  table 
I-D-1.  The  problems  with  pretreatment  are  that  thev 
require  energy,  eciuipment,  and  often  chemu  als. 
they  result  in  an  irretrievable  loss  of  sugar,  and  in 
undesirable  side-reactions  and  byproduct  lorm.i 


Appendix  l-D—The  Impact  of  Genetics  on  Ethanol— A Case  Study  • 295 


Figure  I D-2.  Alternative  Schemes  for  Acid  Hydrolysis  of  Cellulosic  Biomass  for  Ethanol  Production 


Acid 


Acid 


Cellulosic 

biomass 


SOURCE;  Massachusetts  Institute  of  Technology. 


tion.  Furthermore,  if  acids,  alkali,  or  organic  chem- 
icals are  used,  they  must  be  recycled  to  minimize 
cost  or  disposed  of  in  order  to  prevent  pollution. 

In  starch  processing,  prior  to  ethanol  fermenta- 
tion, mechanical  grinding,  steam,  and  enzymes  are 
employed.  The  energ\'  requirements  are  small  and 
contribute  relatively  little  to  the  final  ethanol  cost. 
The  objecth  e in  the  development  of  cellulose-based 
processes  should  be  to  minimize  both  energy  and 
chemical  requirements.  The  development  and  scale- 
up  of  effective  pretreatment  technology  are  under 
acth  e investigation^  and  require  continued  financial 
support  to  better  de\  elop  se\  eral  alternati\  e routes. 
The  most  promising  routes  are:  steam  treatment,  sol- 
\ ent  delignification,  dilute  acid,  cellulose  dissolution, 
and  direct  fermentation. 

Se\eral  different  acid  hydrolysis  schemes  ha\e 
been  proposed.  Ho\ve\er,  most  appear  as  in  flow 
scheme  A or  B in  figure  I-D-2.  Dilute  acid  is  used  to 
hydrolyze  the  hemicellulose  to  pentose  sugars  pri- 
marily and  then  stronger  acid  at  higher  tempera- 

^Proceedings  of  3rd  Annual  Biomass  Energy  System  Conference,  National 
Technical  Information  Service,  SERI  TP-33-285,  1979. 


tures  is  used  to  cause  cellulose  hydrolysis  (scheme 
A).  A major  problem  with  this  approach  is  the  irre- 
\ ersible  loss  of  sugars  to  undesirable  side-product 
formation.  After  separation  of  residual  solids  (mostly 
lignin),  which  can  be  burned  to  provide  energy  for 
distillation,  the  sugar  solution  is  fermented  by  yeast 
to  ethanol.  The  pentose  sugars  also  can  be  fer- 
mented, but  by  organisms  other  than  the  ethanol 
producing  yeast,  to  other  chemicals,  some  of  which 
could  be  used  as  fuels  (e.g.,  ethanol,  acetic  acid, 
acetone,  butanol,  2,3-butanediol,  etc.). 

An  alternative  (scheme  B,  figure  I-D-2)  to  the  above 
is  to  use  a solvent,  after  pentose  sugar  removal,  to 
dissolve  the  cellulose,  allowing  its  separation  from 
lignin.  This  cellulose  solution  is  easily  and  efficiently 
hydrolyzed  to  sugars.  The  advantage  of  this  ap- 
proach over  the  direct  acid  hydrolysis  is  that  the 
yield  of  sugar  is  much  higher.  In  the  harsh  acid  hy- 
drolysis, considerable  sugar  is  destroyed.  However, 
the  major  disadvantage  of  both  these  schemes  is  that 
they  require  recycling  or  disposal  of  acids  and 
solvents.  A second  problem  is  that  almost  nothing  is 
known  about  how  to  scale-up  some  of  the  newly  de- 


296  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Figure  l-D-3.— Alternative  Schemes  for  Enzymatic  Hydrolysis  of  Cellulosic  Biomass  for  Ethanol  Production 


Cellulosic 

biomass 


SOURCE:  Massachusetts  Institute  of  Technology. 


Table  1-0-1.— Alternative  Pretreatment  Methods  for 
Lignocellulose  Materials 


Chemical  methods 

Physical  methods 

Sodium  hydroxide  (alkali) 

Steam 

Ammonia 

Grinding  and  milling 

Chemical  pulping 

Irradiation 

Ammonium  bisulfite 

Freezing 

Sulfite 

Sodium  chlorite 

Organic  solvents 

Acids 

SOURCE:  Office  of  Technology  Assessment. 


veloped  technology,  such  as  that  developed  by 
groups  working  at  Purdue  University,  New  York  Uni- 
versity, and  Dartmouth  College.  There  are  several 
engineering  problems  involving  both  heat  and  mass 
transfer  and  acid/solvent  recycle  that  need  to  be  eval- 
uated at  larger  scale.  At  least  some  of  this  work  will 
be  done  at  the  process  development  unit  now  being 
built  at  the  Georgia  Institute  of  Technology.  The 


most  promising  directions  that  need  de\  ('lopmeni 
are: 

• the  scale-u|)  of  high  ratcts  and  high  yield  labora- 
tory hydrolysis  systems,  and 

• the  developnuMit  of  methods  for  acid  and  chem- 
ical recycle  schttiiMfs. 

There  are  three  ty|Hts  of  approaches  that  ha\e 
been  employed  for  enzymatic  Indrolysis  of  ( (“IIuIomc 
biomass.  These  artf  summarized  in  figure  l-D-.T  They 
all  involve  some  initial  size  reduction  to  increase  the 
surface  area  available  for  enzymatic  attack  In 
schemes  A and  IT  the  incoming  cellulosic  hiom.tss  i;» 
split  into  two  streams;  out'  is  used  to  grow  organisms 
that  produce  cellulolytic  enzymt's  called  eellul.isi-.'- 
and  the  other  is  used  to  produce  sugar. 

In  scheme  A,  tint  eellulases  are  reeo\ cred  and  then 
added  to  a sc[)arate  enzyme  ludrohsis  reat  lion 
They  hydi'olyztf  both  the  cellulose  and  hemieellulosc. 
and  the  resulting  sugar  solution  is  then  p.issed  to  an 
ethanol  fermentation  stage  w hei  e hexoses  are  eon 
verted  by  a yeast  fermentation  to  ethanol  I lili/alion 
of  the  ptfiilose  ri'ciuires  a separate  lermenlalion  lie 


Appendix  l-D— The  Impact  of  Genetics  on  Ethanol— A Case  Study  • 297 


sidiuil  lignin,  which  is  remoxed  hetore  (bv  sohents 
extraction)  or  alter  In  clroh  sis,  is  used  to  pi  o\  ide 
energy  for  ethanol  recoxerv.  Kxtensixe  xvork  on  this 
approach  has  been  done  at  the  I'nix  ersity  of  Califor- 
nia, Berkelex , and  the  I'.S.  .Army  Xatick  Laboratories. 

In  scheme  B.  the  cellulase  is  not  recoxered  but 
rather,  the  xx  hole  fermentation  broth  from  cellulase 
production  is  added  to  the  cellulosic  biomass  along 
xx  ith  ethanol-[)roducing  yeast.  The  result  is  a simul- 
taneous cellulose  hydrolysis  (saccharification)  and 
fermentation.  (In  the  [)roduction  of  ethanol  from 
starch,  the  starch  hydrolyzing  enzymes  are  added  at 
the  same  time  as  the  yeast  for  simultaneous  sacchari- 
fication and  fermentation.)  I'his  technologx'  has  been 
demonstrated  hx  the  Culf  Oil  Co.  After  fermentation, 
the  ethanol  is  recoxered  and  the  residual  lignin  can 
again  he  used  for  energx  for  distillation.  The  prob- 
lem of  unused  pentose  sugar  still  remains  and  xx  ill  re- 
quire a separate  fermentation  step. 

A third  alternatix  e (scheme  C,  figure  l-D-3)  shoxx  s a 
simpler  approach,  nameix  a direct  fermentation  on 
cellulose.  I'his  approach  has  been  dexeloped  at  the 
Massachusetts  Institute  of  Technology'.  It  utilizes 
bacteria  that  xx  ill  produce  cellulase  to  hydrolyze  the 
cellulose  and  hemicellulose  and  ferment  both  the 
hexose  and  pentose  sugars  to  ethanol  in  a single- 
stage  reactor.  The  adxantage  of  this  approach  is  a 
minimal  requirement  for  pretreatment,  a combined 
enzyme  production,  cellulose  hx  drolysis  and  ethanol 
fermentation,  and  simultaneous  conxersion  of  both 
pentose  and  hexose  sugars  to  ethanol.  This  concept 
is  nexx  and  xx  ork  still  needs  to  he  done  to  increase  the 
ethanol  concentration,  minimize  side  product  forma- 
tion, and  increase  the  rate  of  ethanol  production. 
.Again,  residual  lignin  xxill  be  used  to  proxide  the 
energy  for  ethanol  distillation. 

FERME.NT.ATION  OF  ETHAXOL 

.An  examination  of  the  economics  for  ethanol  pro- 
duction shoxx  s that  the  dominant  cost  is  the  process 
raxx  material.  .As  seen  in  table  I-D-2  the  feedstock  rep- 
resents 60  to  70  percent  of  the  manufacturing  cost. 
Thus,  it  is  clear  that  any  improxement  in  substrate 
utilization  efficiency  is  of  substantia]  benefit.  The 
theoretical  yields  of  ethanol  from  glucose,  sucrose, 
and  starch  or  cellulose  are  0.51,  0.54  and  0.57  gram 
(g)  ethanol'g  material,  respectixely;  the  differences 
result  from  the  addition  of  a molecule  of  xvater  on 
hydrolysis.  There  are  sex  eral  approaches  to  improve 
the  yield  abox  e the  typical  value  of  90  to  95  percent 
currently  achiex  ed.  These  are: 

• increase  the  ratio  of  ethanol  produced  per  unit 
weight  of  cells,  e.g.,  through  cell  recycle, 
vacuum  fermentation,  immobilized  cells,  or  im- 
proxement in  specific  productix  ity  (g  ethanol/g 


Table  I-D-2.— A Comparison  of  the  Distribution  of 
Manufacturing  Costs  for  Several  Ethanol 
Production  Processes 


Substrate 

ivlolasses 

Corn 

Grain 

Sorghum 

Cost  component  (%) 

Capital 

9 

12 

10 

Operating 

20 

26 

30 

Feedstock 

71 

62 

60 

Total 

100 

100 

100 

Cost  on  energy  basis 

(SMiVIBtu) 

12.5 

14.9 

12.7 

Cost/gal  etiianol  ($/gal)  . . . 

1.05 

1.25 

1.07 

Capital  investment 

($/annual  gal) 

1.02 

1.05 

1.75 

SOURCE:  "Comparative  Economic  Assessment  of  Ethanol  From  Biomass,” 
Mitre  Corp.,  report  HCP/ET-2854). 


cell  hr),  by  increasing  the  content  and/or  activi- 
ty of  those  enzymes  in  the  pathway  to  ethanol; 

• increase  the  utilization  of  other  materials  in  the 
substrate,  e.g.,  the  use  of  oligosaccharides,  espe- 
cially branched,  in  starch,  and  the  use  of  con- 
taminating sugars  such  as  galactose  or  mannose 
for  hemicellulose;  and 

• dex  elop  a route  for  the  utilization  of  pentose  su- 
gars, especially  xylose,  present  in  hemicellulose. 

The  potential  effect  of  oligosaccharides  or  con- 
taminating sugar  utilization  is  relatively  small,  since 
they  represent  typically  1 to  3 percent  of  the  total 
sugar  content.  Hoxvex'er,  if  cellulosic  biomass  con- 
taining 15  to  25  percent  hemicellulose  is  used,  then 
the  impact  of  pentose  conversion  to  ethanol  is  great. 

Cellulosic  biomass  is  made  up  primarily  of  cellu- 
lose, hemicellulose  (mostly  xylan)  and  lignin.  Other 
components  such  as  protein,  ash,  fats,  etc.,  typically 
comprise  about  10  percent.  The  composition  of  l-io- 
mass  can  be  expressed  in  terms  of  the  following 
equation: 


'-L\ 

where  F^,  F„,  Fl,  and  F^  are  the  weight  fractions  of 
cellulose,  hemicellulose,  lignin,  and  ash,  respectively. 
Assuming  that  the  ash  is  10  percent  (F^  = 0.1)  and 
that  Fj.  and  F„  are  the  only  fermentable  components 
in  the  biomass,  then: 

Fc  = Fh  = 0.9  - Fl  (2) 

The  maximum  amount  of  ethanol  from  one  unit  of 
biomass  (’Ve,b)  is: 

~ ^E/H^H  ~ '^E/B 

Where  and  Ye,h  are  the  yield  of  ethanol  for  cel- 
lulose and  hemicellulose,  respectively.  Equation  2 
can  be  rearranged  to  relate  the  fractions  of  cellulose: 


298  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


Fc  = 0.9  - Fl  - F„  (4) 

Substituting  this  into  equation  3 gives: 

'^EIR  ^E/H^H 

From  equation  5,  the  effect  can  be  calculated  of 
hemicellulose  content  and  conversion  yield  on  the 
overall  conversion  of  biomass  to  ethanol.  Assuming  a 
lignin  content  of  15  percent  (F^  = 0.15)  and  using  Y^/c 
= 0.57  g/g  the  following  equation  is  obtained: 

Ye,b  = 0.43  + Fh(Ye,h  - 0.57)  (6) 

The  theoretical  yield  value  on  hemicellulose, 
Ye,h,  is  not  well-defined  because  so  little  is  known 
about  the  biochemistry  of  anaerobic  pentose 
metabolism.  If  one  mole  of  ethanol  is  produced  per 
mole  of  xylose,  the  yield  is  0.3  g ethanol/g  xylose.  It 
two  moles  of  ethanol  could  be  obained,  Y^/h  would 
be  0.61;  however,  neither  the  mechanism  nor  the 
thermodynamics  of  the  conversion  is  sufficiently 
well-defined  to  allow  one  to  expect  this  value.  The 
maximum  observed  values  are  about  0.41  g 
ethanol/g  xylose.®  The  sensitivity  of  the  overall 
yield  to  this  value  is  shown  in  figure  I-D-4.  The  im- 
pact of  pentose  utilization  depends  on  the  amount 


^S.  D.  Wang  and  C.  Cooney,  Massacliusetts  Institute  of  Technology,  un- 
published results. 


Figure  l•D•4.— Effect  of  Pentose  Yield  on 
Overall  Yield  of  Ethanol  from  Cellulosic  Biomass 
(Ye/b)  with  Varying  Fractions  of  Hemiceilulose  (Fh). 


SOURCE:  Massachusetts  Institute  of  Technology. 


of  hemicellulose  present.  From  the  value  in  figure 
I-D-4  and  the  observation  that  70  percent  of  the 
manufacturing  cost  is  the  raw  material  cost,  it  is 
possible  to  estimate  tbe  economic  benefit  of  pen- 
tose utilization.  Equation  7 relates  the  overall 
ethanol  yield  to  the  manufacturing  cost: 

Cy  = X M (7) 

Ye®  C-7 

where  is  the  manufacturing  cost  per  gallon  of 
ethanol,  Cg  is  biomass  cost  (cents/lb),  6.6  is  the  con- 
version from  pound  to  gallon  of  ethanol,  and  0.7  is 
the  70-percent  factor  for  relative  biomass  cost  to 
ethanol  cost.  For  a biomass  costing  2 cents/lb  and 
containing  20  percent  hemicellulose,  the  manufac- 
turing cost  is  reduced  from  59  to  43  cents/gal,  when 
the  yield  on  pentose  goes  from  zero  to  0.6. 

At  the  present  time,  there  are  few  organisms  that 
produce  more  than  one  mole  of  ethanol  per  mole  of 
pentose  and  none  of  the  usual  alcohol  producing 
yeasts  will  ferment  pentoses  to  ethanol.  Addition  or 
improvement  of  the  ability  to  use  pentose  will  ha\  e a 
major  impact  on  the  economics  of  ethanol  produc- 
tion. 

The  second  major  cost  in  ethanol  production  re- 
lates to  the  cost  of  operation.  Typically,  20  to  30  per- 
cent of  the  final  manufacturing  cost  is  accounted  for 
by  the  sum  of  labor,  plant  o\  erhead,  administration, 
chemical  supplies,  and  fuel  costs.  The  chemical  suf)- 
plies  represent  less  than  1 cent/gal  ethanol  and  may 
be  neglected.  Tbe  labor,  overbead,  and  marketing 
costs  vary  with  plant  size,  but  represent  11  to  7 
cents/gal  for  a 20  to  100  million  gal/yr  plant,  res[)ec- 
tively.  Any  improvement  in  tbe  reduction  of  plant 
size  or  complexity  will  reduce  this  cost;  howe\  (‘r.  the 
economic  impact  is  small.  Fhe  major  component  of 
the  operating  cost  is  the  fuel  charge  for  plant  op(*ra- 
tion  and  for  distillation.  Plant  operations,  eg.,  mix- 
ing, pumping,  sterilization,  starch  gelatinization, 
biomass  grinding,  etc.,  represent  about  20  to  30  per- 
cent of  the  energy  cost.  The  remainder  is  for  ethanol 
distillation  and  residual  solids  drying.  Considerable 
effort  has  been  focused  on  methods  to  impnnc  the 
energy  efficiency  of  distillation  to  reduce  it  from  the 
160,000  Btu/gal  required  for  hexerage  alcohol.  While 
considerable  differences  in  opinion  exist  as  to  the 
minimum,  a reasonable  e.x[)('ctation  is  about  40.000 
Btu/gal  although  current  technology  retiuires  6!). 000 
Btu/gal.®  Forty  thousand  Btu  is  about  half  of  the  ener- 
gy content  of  ethanol  per  gallon. 

A discussion  of  process  imiiroxcments  relating  to 
ethanol  recovery  has  two  coni|)on('nts  I he  first  is 

"Report  of  the  Casohol  Sliulv  Croup  ol  Ihe  f.nerj^v  He  rar<  h \d\i»or\ 
Board,  Deparlnienl  of  Euiei'tw.  U d!ihin«lon  1)  ( *\l  (,ilii.  .out  H 

D DeMo.ss,  "Klhanol  Formation  m I’srinlomonHf  lindnrr;  \r,  h n - ' •' 

Biophys.  34:47H-479,  I9.X1 


Appendix  l-D—The  Impact  of  Genetics  on  Ethanol — A Case  Study  • 299 


related  to  operating  costs  and  the  second  is  related  to 
energ\  et'ticiencv . It'  coal  is  used  to  [)ro\  ide  energ\' 
tor  distillation,  and  it  is  valued  at  S30/ton,  with 
10,51)0  Btu  Ih  or  S 1 .50/million  Htu,  then  the  energy 
cost  for  distillation  (optimistically  assuming  40,000 
Btu  gal)  is  SO  gal.  If  lignin  from  cellulosic  hiomass  is 
used  as  a fuel,  the  cost  is  reduced  further.  On  the 
other  hand,  if  oil  at  $40,1)hl  (130,000  Btu/gal  and  42 
gall)!)!)  or  S7  million  Btu  is  used,  then  the  energv' 
cost  is  28  cents  gal  of  ethanol. 

From  a common  sense,  economic,  and  political 
point  of  view,  it  does  tiot  seem  reasonable  to  utilize 
liquid  fuel  to  produce  liciuid  fuel  from  hiomass. 
rherefore,  it  w ill  he  assumed  that  petroleum  will  not 
he  used  for  distillation  and  that  either  coal  or  bio- 
mass will  lie  employed. 

In  order  to  assess  the  impact  of  process  improve- 
ments on  the  energv  demand,  it  is  necessary  to  look 
at  an  o\  erall  material  balance.  This  is  summarized  in 
figure  I-I)-5.  Only  a portion  of  the  entering  biomass 
feedstock  is  fermented  to  ethanol  and  there  are  two 
product  streams,  one  containing  ethanol  and  the 
other  solids,  both  must  he  separated  from  water.  It  is 
important  to  note  that  as  the  ethanol  concentration  is 
increased,  the  energv  requirement  for  both  ethanol 
recovery  from  the  water  and  for  drying  will  de- 
crease. Therefore,  the  impact  of  developing  ethanol 
tolerant  micro-organisms  is  seen  as  a reduction  in 
energv’  cost. 

Figure  l-D-5.— Process  Schematic  for  Material  and 
Energy  Balance 


Biomass 


Solids 


SOURCE;  Massachusetts  Institute  of  Technology. 


The  third  major  cost  for  ethanol  manufacturing  is 
the  capital  investment,  which  represents  about  4 to 
12  percent  of  the  manufacturing  cost.  The  capital  in- 
vestment is  determined  by  the  complexity  of  the 
processes  and  the  volumetric  productivity  of  ethanol 
production.  Thus,  the  development  of  a micro-orga- 
nism that  will  require  a minimum  amount  of  feed- 
stock pretreatment  and  will  produce  ethanol  at  a 
higher  rate  will  reduce  the  net  capital  investment. 

The  volumetric  productivity  (Q^)  for  ethanol  pro- 
duction is  given  by: 

Qe  = 

where  q^  is  the  specific  productivity  expressed  in  g 
ethanol  per  g cell  hr,  and  X is  the  culture  density. 
Therefore,  there  are  two  approaches  to  obtain  high 
productivity;  first,  to  choose  or  create  an  organism 
with  a high  specific  rate  of  ethanol  production  and 
second,  to  design  a process  with  high  cell  density. 

The  application  of  genetics  can  be  used  to  enhance 
the  intracellular  enzyme  activity  of  the  enzymes 
used  for  ethanol  production.  The  resulting  increase 
in  Qp  will  result  in  reduced  capital  investment  re- 
quirements. 

There  are  four  types  of  ethanol  processes  based 
on  different  organisms;  they  are: 

1.  Saccharomyces  cerevisiae  and  related  yeast, 

2.  Saccharomyces  cerevisiae/T richoderma  reesei, 

3.  Zymomonas  mobilis,  and 

4.  Clostridium  thermocellum/thermosaccharolyti- 
cum,  or  thermohydrosulfuricum. 

The  first  is  the  traditional  yeast  based  process  using 

S.  cerevisiae  to  ferment  soluble  hexose  sugar  to  eth- 
anol. In  the  second,  the  substrate  range  is  extended 
to  cellulose  by  the  use  of  cellulase  produced  by  T. 
reesei.  The  third  approach  utilizes  Z.  mobilis;  this 
organism  is  a particularly  fast  and  high  ethanol  yield- 
ing one.  Its  range  of  fermentable  substrates,  how- 
ever, is  limited  to  soluble  hexose  sugars. 

In  many  tropical  areas  of  the  Americas,  Africa, 
and  Asia,  alcoholic  bev^erages  prepared  from  a mixed 
fermentation  of  plant  steeps  are  popular.  Bacteria 
from  the  genus  Zymomonas  are  commonly  em- 
ployed. In  the  early  1950's,  the  genus  Zymononas  ac- 
quired a certain  fame  among  biochemists  by  the  dis- 
covery that  the  anaerobic  catabolism  of  glucose 
follows  the  Enter-Doudoroff  mechanism.^  This  was 
very  surprising,  since  Zymomonas  was  the  first  ex- 
ample of  an  anaerobic  organism  using  a pathway 
mainly  in  strictly  aerobic  bateria.® 

In  spite  of  its  extensive  use  in  many  parts  of  the 
world,  its  great  social  implications  as  an  ethanol  pro- 

=M.  Gibbs  and  R.  D.  de  Moss,  "Ethanol  Formation,  in  Psuedomonas 
Undneri,"  Arch.  Biochem.  Biophys..  34:478-479,  1951. 

®J.  Swings  and  J.  DeLey,  "T  he  Biology  of  Zymomonas,"  Bacteriological  Re- 
views 41:1-46,  1977. 


300  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


clucer,  and  its  unique  biochemical  position,  Zymo- 
monas  has  not  been  studied  extensively/ 

The  organism  most  often  studied  is  Zymomonas 
mobilis,  which  can  produce  up  to  1.9  moles  of 
ethanol  per  mole  of  glucose.  Recent  studies  reported 
from  Australia,  have  established  the  Z.  mobilis  can 
ferment  high  concentrations  of  glucose  rapidly  to 
ethanol  in  both  batch  and  continuous  culture  with 
higher  specific  glucose  uptakes  rates  for  glucose  and 
ethanol  production  rates  than  for  yeasts  currently 
used  in  alcohol  fermentations  in  Australia.®  ® 

For  example,  several  kinetic  parameters  for  a Z. 
mobilis  fermentation  were  compared  with  Saccha- 
romyces  carlsbergensis^^'  specially  selected  for  its 
sugar  and  alcohol  tolerance. “ Both  specific  ethanol 
productivity  and  specific  glucose  uptake  rate  are  sev- 
eral times  greater  for  Z.  mobilis.  This  result  is  mainly 
due  to  lower  levels  of  biomass  formation  and  glucose 
consumption.  The  lower  biomass  produced  would 
seem  to  be  a consequence  of  the  lower  energy  avail- 
able for  growth  with  Zymomonas  than  with  yeasts— 
the  Enter-Doudoroff  pathway  producing  only  1 mole 
of  adenosine  triphosphate  (ATP)  per  mole  of  glucose, 
compared  to  glycolysis  with  2 moles  ATP  per  mole 
glucose.  In  none  of  the  first  three  examples  can  etha- 
nol be  produced  from  pentose  sugar. 

The  fourth  approach  utilizes  a mixed  culture  of 
Clostridia,  which  will  utilize  cellulose  and  hemicellu- 
lose,  hexoses,  and  pentoses  for  ethanol  production. 

The  application  of  genetics  for 
improving  microbial  strains 

In  the  previous  sections,  the  process  steps  have 
been  identified  that  are  particularly  sensitive  to  the 
quality  of  the  microbial  strains.  The  following  are  im- 
provements of  microbial  characteristics  that  are 
either  now  possible  or  might  be  so  in  the  future  and 
that  will  have  an  impact  on  the  overall  economics  of 
the  process.  The  effect  of  new  genetic  techniques  re- 
quiring future  research  is  similar  for  all  micro-orga- 
nisms in  two  ways. 

1.  Manipulations  could  be  attempted  today  with 
less  effort  and  greater  chance  of  success  if  tools 
like  cell  fusion  and  recombinant  DNA  (rDNA) 
techniques  were  available  for  all  of  the  mi- 
crobes of  interest. 


'Gibbs,  et  al.,  op.  cit. 

»K.  J.  Lee,  D.  E.  Tribe,  and  P.  L.  Rogers,  "Ethanol  Production  by  Zymo- 
monas mobilis  in  Continuous  Culture  at  High  Glucose  Concentrations,"  Bio- 
technology Lett . 421-426,  1979. 

®P.  L.  Rogers,  K.  J.  Lee,  and  D.  E.  Tribe,  Biotechnol.  Lett.  1:165-170,  1979. 
'“Ibid. 

”D.  Rose.  Proc.  Bichem.  1 1(2),  1976,  pp.  10-12. 


2.  Manipulations  require  further  knowledge  in  a 
specific  area  or  the  development  of  an  entirely 
new  genetic  system  in  ethanol  producing  mi- 
crobes—e.g.,  there  is  no  genetic  system  for  the 
thermophilic  anaerobic  bacteria.  Knowledge  on 
how  to  genetically  alter  ethanol  tolerance  of 
both  bacteria  and  yeast  is  lacking. 

The  economics  of  the  fermentation  of  a substrate 
into  alcohol  is  primarily  controlled  by  three  factors: 

1.  Ethanol  yield.— The  amount  of  product  pro- 
duced per  unit  of  substrate  determines  the  ma- 
jor raw  materials  cost  of  the  fermentation. 

2.  Final  ethanol  concentration.— The  cost  of  separat- 
ing the  ethanol  from  the  fermentation  broth  is  a 
function  of  the  ethanol  concentration  in  that 
broth. 

3.  Productivity.— The  amount  of  ethanol  produced 
per  liter  of  fermenter  \olume  per  hour  deter- 
mines the  capital  cost  of  the  fermentation  step, 
once  the  type  of  fermenter  and  the  annual  out- 
put have  been  chosen.  Productivity  is  not  inde- 
pendent of  the  final  ethanol  concentration,  and 
so  an  optimum  compromise  between  these  vari- 
ables must  be  chosen. 

The  impact  of  genetics  on  ethanol  yield 

Most  microbes  that  are  chosen  for  making  ethanol 
already  produce  nearly  the  theoretical  maximum 
yield.  In  these  cases  little  improvement  can  he  made. 

The  yield  may  he  lower  when  the  microbe  has 
been  chosen  for  its  other  technical  advantages  such 
as  ability  to  degrade  cellulose,  bower  yield  of  a 
microbial  end  product,  like  ethanol,  can  result  fi’om 
the  diversion  of  substrate  to  cell  mass  or  to  an  alter- 
native product.  Both  of  these  faults  can  he  readily  at- 
tacked. A number  of  cell  changes  (e.g.,  leaky  mem- 
branes) can  cause  the  microbe  to  waste  energ^v,  re- 
quiring it  to  metabolize  more  suhstrati’  into  alcohol 
to  make  the  same  cell  mass.  Where  the  thermo- 
dynamics and  redox  balance  of  the  fermentation 
allow,  unwanted  waste  jiroducts  can  he  eliminated 
by  mutation  of  the  relevant  pathways.  Only  limited 
work  has  been  done  on  this  type  of  research  w ith  in- 
dustrially sigificant  bacteria. 

The  impact  of  genetics  €tn  final  idhantil 
concentration 

This  is  amenable  to  genetic  manipulation,  both  em- 
pirical and  planned.  An  impro\ement  in  ethanol  tol- 
erance decreased  both  separation  costs  and  ferment- 
er capital  cost  (through  increased  productiv  ilyl 

When  traditional  distillation  is  used,  the  ellei  t on 


Appendix  l-D— The  Impact  of  Genetics  on  Ethanol— A Case  Study  • 301 


the  separation  cost  of  increased  ethanol  tolerance  is 
smaller  once  ethanol  concentrations  ha\e  reached 
approximately  6 percent.  Howexer,  the  importance 
of  increased  ethanol  concentration  to  fermenter  pro- 
ductix  ity  remains. 

It  is  likely  that  the  most  important  inhihitorv  ac- 
tion ot  ethanol  takes  place  at  the  cell  membrane. 
Strategies  for  manipulating  the  cell  membrane  com- 
position and  properties,  and  understanding  in  this 
area,  are  increasing  rapidly. 

i Genetics  and  ethanol  tolerance 

I 

The  study  of  ethanol  tolerance  by  micro-orga- 
nisms has  been  approached  using  strains  with 
altered  genetic  makeup.  Sex  eral  kinds  of  Escherichia 
coli  mutants  hax  e been  isolated  hax  ing  different 
tolerances  to  ethyl  alcohol.'-  Solxent  resistant  strains 
either  had  larger  amounts  of  total  phospholipid  (type 
III)  or  had  an  altered  phospholipid  and  membrane- 
hound  protein  composition  (type  II).  On  the  other 
hand,  mutants  with  a lesion  mapping  close  to  pss 
gene  (which  codes  for  phosphotidylserine  syn- 
thetase) were  either  solx  ent  sensitixe  or  resistant.'^ 

The  physiologx-  of  an  E.  coli  ethanol  resistant  mu- 
tant has  been  characterized  similarly.'^  This  strain 
had  pleiotropic  groxx  th  defects  including  abnormal 
cell  dix  ision  and  morphologx'.  It  also  had  an  altered 
tac  permease  that  x\  as  not  due  to  a mutation  in  the  V 
gene.  It  xxas  concluded  that  altered  membrane  com- 
position xx  as  responsible  for  this  abnormal  behax  ior. 

More  recently,  ethanol  tolerant  mutants  hax  e been 
isolated  from  C.  thermocellum.^^  Indirect  exidence 
lead  to  the  conclusion  that  strain  S-4  xx  as  defectix  e in 
hydrogenase,  since  this  strain  produced  loxx^er 
amounts  of  acetic  acid.'®  A different  ethanol  resistant 
isolate  of  the  same  bacterium,  strain  C9,  proved  to 
hax  e a loxver  actu  ation  energx-  for  groxx  th  than  the 
xvild  type,  a property  that  has  been  related  to  mem- 
brane composition. 

There  are  three  categories  of  changes  that  could 
influence  the  fermentation  process: 

1.  Manipulate  the  existing  controls  on  metabolism. 

Consider  an  example.  In  many  organisms  the 


'-D.  P.  Clark  and  J.  P.  Beard.  ".Altered  Phospholipid  Composition  in  .Mutants 
of  Escherichia  Coli  Sensitive  or  Resistant  to  Organic  Solvents."  J.  Gen. 
Microbiol.  113:267-274,  1979. 

'^.A.  Ohta  and  I.  Shibuva,  'Membrane  Phospholipid  Synthesis  and  Pheno- 
typic Correlation  of  an  E.  Coli  pss  -Mutant,"  J.  Bacteriol.  132:434M43,  1977. 

“X  . .A.  Fried  and  A.  Xovick,  "Organic  Solvents  as  Probes  for  the  Structure 
and  Function  of  the  Bacterial  Membrane:  Effects  of  Ethanol  on  the  XX  ild 
T\pe  and  as  Ethanol  Resistant  -Mutant  of  Escherichia  Coli,"  J.  Bacteriol. 
114:239-248.  1973. 

•®S.  D.  XX  ang,  "Production  of  Ethanol  From  Cellulose  by  Clostridium  Ther- 
mocellum,  .M  S.  Thesis,  Department  of  -Nutrition  and  Food  Science,  Massa- 
chusetts Institute  of  Technology,  1979. 

'Mbid. 


energy'  level  of  the  cell,  expressed  through 
adenosine  monophosphate  (AMP),  adenosine  di- 
phosphate (ADP),  and  adenosine  triphosphate 
(,ATP)  levels,  partially  controls  the  rate  of  gly- 
colysis. A defective  cell  membrane  xvould  pro- 
xide  an  energy  sink,  to  keep  glycolysis  at  its 
maximum  rate.  Strategies  such  as  this  could  be 
attempted  noxv. 

2.  Increase  the  amount  of  each  transport  and  cata- 
bolic enzyme  in  the  fermentation  pathway.  This 
requires  the  ability  to  isolate  the  genes  of  in- 
terest and  to  amplify  them  xvith  in  vivo  or  in 
x itro  recombinant  techniques  in  the  microbe  of 
interest.  This  is  not  an  immediate  prospect. 

3.  Accomplish  complete  deregulation  of  the  fer- 
mentation pathxvay  in  the  microbe  of  interest. 
Essential  catabolic  enzymes  are  difficult  to 
manipulate,  and  this  is  also  not  an  immediate 
prospect. 

Genetic  manipulation  of  the  microbe  can  influence 
fermentation  processes  in  other^  ways  as  well.  These 
are  less  important  than  improvements  in  yield,  final 
ethanol  concentration,  and  productivity,  but  they 
also  affect  the  cost.  Examples  are: 

• tx'pe  of  fermenter  used; 

• nonsubstrate  nutrients; 

• strain  stability; 

• cell  separations  for  byproducts,  recycle,  or  eth- 
anol recovery  (i.e.,  increased  size  for  recovery); 

• operating  conditions,  i.e.,  higher  groxvth  tem- 
peratures for  yeast  and  mesophilic  bacteria;  and 

• range  and  efficiency  of  substrate  utilization  (i.e., 
complete  utilization  of  all  sugars). 

More  detailed  examples  are: 

• Type  of  fermenter.— If  the  organism,  whether  it 
be  a yeast  or  a bacterium,  can  be  made  to  grow 
under  conditions  of  pH,  ethanol  concentration,  tem- 
perature, etc.,  that  preclude  contamination,  inexpen- 
sive lined  basins  can  be  used  instead  of  tanks,  since 
steam  sterilization  of  the  fermenter  is  not  required. 
In  this  case,  some  operating  and  capital  costs  asso- 
ciated xvith  sterilization  are  avoided  as  well. 

A type  of  continuous  beer  fermenter  requires 
groxxth  in  the  form  of  fast-settling  pellets.  In  other 
fermenters,  fast-settling  particles  (such  as  mycelia) 
present  problems  that  are  best  avoided  by  agglom- 
eration of  the  cell  mass.  This  type  of  control  over  the 
growth  form  of  micro-organisms  is  amenable  to 
genetic  manipulations. 

• Nonsubstrate  medium  costs.— In  addition  to  the 
carbon-energy  substrate  and  water,  growing  cells 
must  be  supplied  with  other  nutrients.  Some  orga- 
nisms can  make  all  of  their  biochemicals  from  quite 
simple  sources  of  nitrogen,  phosphorus,  sulfur, 
magnesium  and  trace  metals.  Others  require  more 


302  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


complex  molecules,  ready-made,  such  as  amino  acids 
and  vitamins. 

The  more  cheaply  these  nutrient  needs  can  he 
provided,  the  better.  Whenever  an  organism  can  be 
given  genes  from  another  source  by  applied  biotech- 
nology techniques,  there  is  a possibility  that  complex 
nutrient  requirements  can  be  obviated.  However, 
this  requires  that  all  the  genes  in  a given  pathway  be 
located  in  the  source  and  be  made  to  function  in  the 
new  microbes.  The  feasibility  of  this  is  uncertain,  but 
solutions  would  decrease  the  cost  of  producing  etha- 
nol with  yeast  as  well  as  Clostridia. 

• Stain  stability.— Many  of  the  suggested  ethanol 
processes  propose  to  employ  continuous  culture. 
Although  this  offers  several  advantages  over  batch 
culture,  it  is  somewhat  vulnerable  to  deleterious  mu- 
tations of  the  microbe  used,  particularly  if  the  mi- 
crobe has  been  extensively  altered  in  ways  that  make 
it  less  competitive. 

These  deleterious  genetic  changes  are  almost  en- 
tirely catalysed  by  biological  systems  in  the  microbe. 
Alteration  of  these  systems,  so  that  the  frequency  of 
unwanted  genetic  changes  is  decreased,  could  great- 
ly extend  the  period  of  operation  that  is  possible 
before  having  to  shut  down  and  restart  the  fermen- 
tation. So  far,  this  is  a possibility  only  in  microbes 
that  have  a highly  developed  genetics.  It  may  be  that 
strain  stabilization  of  this  sort  would  not  be  possible 
in  other  microbes  until  their  genetics  are  highly  de- 
veloped. 

It  is  also  possible  to  design  strategies  using  current 
strain  development  techniques  that  might  lead  to 
genetically  stable  strains,  but  these  are  unproven. 

• Cell  separations.— Many  fermentation  schemes 
incorporate  cell  recycle  to  boost  productivity.  This 
requires  that  cells  be  separated  from  effluent  broth. 
Others  need  to  separate  cells  from  other  residue  as  a 
byproduct.  In  addition,  some  of  the  low-energy  alter- 
natives to  distillation,  such  as  adsorption,  could  re- 
quire separation  of  the  cells  from  the  broth  prior  to 
ethanol  recovery. 

In  these  cases,  microbes  that  can  be  made  to  floe-  ’ 
culate  and  redisperse,  or  that  can  be  made  to  rever- 
sibly change  their  morphology  would  allow  cheap 
gravity  separations  (settling  or  flotation). 

• Operating  conditions.— An  increase  in  the 
temperature  an  organism  will  tolerate  is  advanta- 
geous for  heat  removal  and  in  situ  ethanol  removal 
schemes.  The  feasibility  of  accomplishing  this  is 
uncertain. 

The  extreme  of  productivity  improvement  via  cell 
recycle  is  an  immobilized  cell  reactor.  It  is  con- 
ceivable that  cells  could  be  made  less  prone  to 
degradation  under  the  conditions  of  immobilization, 
by  modifying  sensitive  components  and  degradation 


systems,  and  by  adding  protective  systems.  This  is 
not  at  all  a near-term  possibility. 

• Range  and  efficiency  of  substrate  utilization.— A 
single-step  conversion  of  a substrate  to  ethanol  is 
highly  desirable.  This  often  requires  that  the  ethanol 
fermenting  organism  possess  a degradation  capabili- 
ty it  does  not  have. 

As  an  example,  consider  ligno-cellulose.  It  consists 
of  hexosans,  pentosans,  and  lignin.  All  of  these  com- 
ponents should  be  used.  Assume  that  one  cellulase- 
producing  candidate  does  not  use  pentoses,  while  a 
related  noncellulase  producing  organism  does,  this  is 
exactly  the  situation  with  clostridia.  If  the  second 
organism  can  be  given  the  cellulase  genes  of  the  first, 
a microbe  better-suited  to  direct  conversion  could  he 
created.  The  pace  at  which  such  a manipulation 
could  be  developed  cannot  be  predicted  with  con- 
fidence, although  this  is  not  necessarily  a long-term 
prospect. 

Another  obvious  area  that  merits  attention  is  the 
enhancement  of  cellulase  activity.  Classical  genetic 
manipulations,  employing  mutation  and  selection  or 
screening,  should  result  in  micro-organisms  better 
equipped  to  degrade  cellulose.  E.g,  it  should  he  possi- 
ble to  isolate  strains  that  are  deregulated  in  cellulase 
production  (hyperproducers)  as  well  as  those  in 
which  the  cellulase  is  not  subject  to  [jroduct  inhibi- 
tion. In  addition,  it  is  tempting  to  think  about  the 
possibilities  of  amplifying  cellulase  genes  by  im'ans 
of  DNA  technology  and  cloning.  How(ner,  this  latter 
approach  must  await  further  understanding  of  the 
biochemistry  and  genetics  of  the  cellulase  .system  as 
well  as  the  development  of  the  a|)pi'opriate  genetic 
systems  in  cellulolytic  micro-organisms. 

Utilization  of  fermentation  byproducts 

Presently  for  each  gallon  of  ethanol  |)rodueed,  ap- 
proximately 14  liters  of  stillage  is  formed.'^  If  ethanol 
is  mixed  with  gasoline  to  make  gasohol  (10  percent 
ethanol),  the  total  stillage  pioduci'd  annually  iti  the 
United  States  would  he  in  tlie  billions  of  liters.  Sui  ('ly 
a problem  of  this  magnitude  d(?sern!s  serious  atten- 
tion. The  utilization  of  stillage  or  ferm(>ntation  by- 
products could  be  greatly  improved  In  genetic 
means  in  several  ways.  In  actuality,  only  a lational 
long-range  genetic  approach  can  increa.se  tin*  value 
of  such  a fermentation  byproduct.  V alue  can  he  in- 
creased in  two  main  ways.  The  fir  st  is  to  increase  the 
nutritive  value  of  the  fermentation  byproduct  fol- 
lowed by  develoiring  economical  processing  technol- 


*^W.  K.  Tyner,  ' The  j^otenlliil  ot  ( )l)l.iinin/'  I .nei  \ mm  \i;i  m ulhi'  • 
posium  on  Biulerhnolof^v:  I'lw  hlnrri^v  Pnuiin  tinn  ntuf  ( tmsri  \ (..iilut 

berg,  Tenn.,  1979. 


Appendix  l-D — The  Impact  of  Genetics  on  Ethanol — A Case  Study  • 303 


ogies  that  stabilize  and  presen  e nutritive  value.  The 
second  approach  is  to  increase  the  functionality  of 
the  byproducts  so  that  more  useful  products  can  be 
developed. 

For  this  one  can  envisage  clever  and  novel  ways  to 
utilize  mutants  to  increase  the  value  in  a manner 
similar  to  those  described.**  **  Ethanol  production 
is  not  compatible  with  producing  a \ aluable  byprod- 
uct. E.g.,  a filamentous  yeast  may  be  useful  for  direct 
te.xturization  or  fortification  of  an  animal  food  but 
production  of  ethanol  may  not  be  suitable  with  such 
an  organism.  .-\  possible  solution  to  this  type  of  con- 
flict in\olves  the  de\elopment  and  engineering  of 
two-stage  fermentation  processes.  In  the  first  stage, 
ethanol  producing  organisms  are  propagated  under 
optimal  economic  conditions  for  ethanol  production. 
.After  the  production  phase  is  over,  the  organisms 
are  then  transferred  to  a second-stage  reactor, 
where  desirable  phenotypic  properties  are  then  e.\- 
pressed.  Signals  for  e.xpression  of  phenotypic  prop- 
erties can  be  extrinsic  environmental  parameters, 
such  as  temperature,  or  levels  of  o.xygen  or  carbon 
dioxide,  or  intrinsic  parameters,  such  as  specific 
nutrient  requirements. 

Thus  the  large-scale  utilization  of  fermentation 
byproducts  as  feed  or  other  materials  will  then 
become  more  valuable  when  genetic  engineering  can 
decrease  processing  costs  and  increase  product 


'•A.  J.  Sinskev,  J.  Boudrant.  C.  Lee.  J.  De.Angelo.  V.  Miyasaka,  C.  Rha.  and  S. 
R.  Tannenbaum,  Applications  of  Temperature-Sensitive  Mutants  for  Single- 
Cell  Protein  Production, " in  Proceedings  of  L'.S./U.S.S.R.  Conference  on  Mech- 
anisms and  Kinetics  of  L’ptake  and  L'tUization  of  Substrates  in  Processes  for 
the  Production  of  Substances  by  Microbiological  Means,  Moscovv-Pushchino, 
p.  362.  June  4-11.  1977.  PB.  283-330-T. 

”J.  Boudrant.  J.  De.Angelo.  A.  J.  Sinskev.  and  S.  R.  Tannenbaum.  "Process 
Characteristics  of  Cell  Lysis  Mutants  of  Saccharomyces  cer\iciae."  Biotech. 
Bioeng.  21:659.  1979. 

=“V.  Miyasaka.  A.  J.  Sinskev,  J.  De.Angelo.  and  C.  Rha,  "Characterization  of 
a Morphological  Mutant  of  Saccharomyces  cer\isiae  for  Single-Cell  Protein 
Production,"  J.  Food Science 45:558:563.  1980. 


quality.  Most  of  these  types  of  studies  remain  to  be 
done.  However,  the  potential  for  innovative  applica- 
tions is  great,  but  such  applications  may  not  result 
because  of  the  current  lack  of  any  Government  agen- 
cy that  has  a sound  program  for  funding  biotech- 
nologA'  research. 

Recommendations  and  areas  in  vrhich 
applied  genetics  should  have  an  impact 

There  has  been  little  published  research  done  in 
the  United  States  on  the  genetic  improvement  of 
ethanol  production  processes  with  bacteria  such  as 
Zymomonas  and  clostridia,  and  only  limited  studies 
with  yeast.  In  light  of  previous  discussion,  the  follow- 
ing points  have  been  identified  as  being  the  most  im- 
portant and  relevant  in  the  application  of  genetics 
for  improving  ethanol-producing  processes: 

• improvements  on  ethanol  yield; 

• increased  ethanol  tolerance  to  achieve  higher 
final  ethanol  concentrations  in  the  fermentation 
broth; 

• increased  rates  of  ethanol  production; 

• elimination  of  unwanted  products  of  anaerobic 
catabolism,  that  is,  direction  of  catabolism 
towards  ethanol; 

• enhanced  cellulolytic  and/or  saccharolytic  capa- 
bilities to  improve  rates  of  conversion  of 
cellulose  and/or  starch  to  fermentable  sugars; 

• incorporation  of  pentose  catabolic  capabilities 
into  ethanol  producers; 

• development  of  strains  capable  of  hydrolyzing 
cellulose  and  starch  as  well  as  of  producing 
ethanol  from  pentoses  and  hexoses; 

• improved  temperature  stability  of  micro-orga- 
nisms and/or  their  enzymes;  and 

• improved  harvesting  properties  of  cellular  bio- 
mass produced  during  fermentation. 


Appendix  II-A 

A Case  Study  of  Wheat 


wheat  is  a major  food  staple  in  the  diet  of  a large 
percentage  of  the  world’s  population.  Wheat  grain  in 
the  United  States  is  used  almost  exclusively  for 
human  consumption,  although  temporary  localized 
oversupply  may  result  in  some  wheat  feeding  to  live- 
stock. 

Attempts  to  improve  wheat  plant  populations  by 
selection  began  several  thousand  years  ago.  The  de- 
sirable attributes  selected  included  the  ability  to 
withstand  severe  environmental  stresses  such  as 
heat,  cold,  and  drought  and  the  stability  of  the  seed 
head  (which  tends  to  disarticulate  in  wild  forms). 

Wheat  seeds  moved  from  country  to  country 
along  with  explorers  and  colonists.  New  varieties 
played  major  roles  in  the  establishment  of  many 
productive  wheat  cultures— e.g.,  the  Mennonite  set- 
tlers introduced  hard  red  winter  (Turkey  Red)  wheat 
into  the  Kansas  area  from  Russia  in  the  late  19th 
century.  And  two  private  breeders— E.  G.  Clark  of 
Sedgenick,  Kans.,  and  Danne  of  Elreno,  Okla.— de- 
veloped varieties  that  set  new  levels  of  productivity 
and  straw  strength  in  hard  winter  wheats  which 
were  sought  by  millers  for  their  excellent  flour 
recovery. 

Breeding  programs  expanded  during  the  first  half 
of  the  20th  century.  At  first,  the  U.S.  Department  of 
Agriculture  (USDA)  played  a lead  role;  but  the 
emergence  of  the  Land  Grant  System  and  the  estab- 
lishment of  the  State  experiment  station  concept 
prompted  individual  States  to  launch  breeding  pro- 
grams designed  to  address  the  particular  production 
problems  faced  by  farmers  within  their  respective 
boundaries. 

As  the  State  experiment  stations  began  to  assume 
more  responsibility,  USDA  programs  and  personnel 
began  to  concentrate  in  central  locations  to  assemble 
the  optimal  number  of  personnel  for  the  greatest  in- 
teraction and  productive  output.  If  the  present  trend 
continues,  there  will  be  virtually  no  USDA  scientists 
engaged  in  actual  wheat  breeding.  Instead  they  will 
have  assumed  the  roles  of  basic  researchers  and  re- 
gional coordinators  supplying  information  to  the 
public  and  private  breeders. 

Disease  and  insect  resistance  have  been  the  pri- 
mary breeding  goals  of  many  programs.  The  dramat- 
ic losses  associated  with  severe  pest  problems  have 
focused  the  attention  of  producers,  researchers,  and 
legislators  on  these  areas  of  need.  Other  traditional 
breeding  objectives  have  included  improved  use 
properties,  tolerance  to  environmental  stresses  such 


as  cold,  wheat,  wind  dessication,  and  excessi\e  mois- 
ture, and  inherent  yield  capacity  in  the  absence  of 
significant  production  limitations. 

The  quality  of  wheat’s  end  products  has  been 
impro\ed  significantly  through  breeding.  Varieties 
have  been  tailored  to  meet  the  demands  of  \ arious 
industries.  The  bread  bakeries  needed  a higher  pi’o- 
tein  and  more  gluten  strength  to  make  a lighter, 
larger  loaf,  while  the  cookie  producer  needed  a low  - 
protein  flour  w ith  desirable  dough-spreading  prop- 
erties. 

Wheat  productivity  and  management 

The  pattern  of  wheat  productiv  ity  (yield  |)er  acre) 
in  developed  countries  is  remarkably  similar.  When 
yields  are  plotted  o\er  the  centuries,  there  is  a long 
period  of  barely  perceptible  increases  in  yield,  from 
the  time  of  first  records  of  production  to  the  end  of 
the  first  third  of  this  century  (the  period  of  1925-35). 
Since  around  1935,  yield  has  increased  sharply.  Re- 
cent data  suggest  that  yield  increases  may  he  U'\  (>ling 
off.  Why  increases  have  been  so  substantial  after 
generations  of  little  success,  is  a complex  ciuestion  in- 
voh'ing  genetic  resources,  economic  de\  ('lo|)m('nt. 
social  interaction,  and  ado|)tion  of  mechanical  and 
biological  inno\  ations. 

Lintil  recently,  the  U.S.  commercial  seed  com|)a- 
nies,  with  one  or  two  exceptions,  ha\e  not  been  in- 
terested in  wheat  breeding  programs  as  a prolitmak- 
ing  venture.  Since  wheat  has  a perfect  flower  and 
can  fertilize  itself,  the  fai  nier  can  |)urchase  seed  ol  a 
new'  variety  and  reproduce  it  from  generation  to 
generation,  llowexer,  the  di.scxncrv  of  cyto|)lasmic 
male  sterility  and  nuclear  restorer  genes  has  stim- 
ulated industry  interest  in  the  possibility  of  devel- 
oping hybrid  wheat.  The  farmer  would  purchase  the 
hybrid  seed  each  yc^ar:  the  inl)red  lines  used  to  make 
the  hybrid  would  he  the  exclusive  |)ropertv  ol  the 
originating  company.  .Although  |)rogress  h.is  been 
good,  problems  (rxisi  with  tin*  sterililv  and  restorer 
systems,  the  ability  to  produce*  ade(|uate  amounts  ol 
hybrid  seed,  and  the  id(*ntification  ol  economie  levels 
of  hybrid  vigor.  The  next  5 years  should  reveal  live 
potential  for  success  in  hybrid  w heal. 

Several  milestone’s  e)f  |)re)gre*ss  have*  he*e*n  se*l  m 
wheat.  Meld  has  rise*n  elramatie  allv  (.e*ne*lie-  preile*e  - 
tion  against  |)ests  anel  e)the*r  ha/arels  has  he*e*n  a m.i 
jor  contiihute)!'  lee  ine  ie*ase*el  yie’lels  In  .lelelilieen  re* 
cent  advances  using  se*mielwarf  ge*ne*s  have*  he*e*n  ,is 


304 


Appendix  ll-A— A Case  Study  of  Wheat  • 305 


sociatrcl  with  signitirant  yield  improv eim'iit.  I'he 
shorter,  stilh'r  stems  ot  the  semidwarl  plants  allow 
ma\imi/ation  ot  ri'sourees  w ithout  yield  l eduetions. 
lm|)ro\ »'ment  in  thi>  inluM’iMit  yield  components  of 
st('ins  p«M-  unit  ar(>a,  kernt'Is  per  stem,  and  kernel 
weif'ht  has  also  contrihuteil  (>\t('nsi\ el\  to  yield  im- 
pro\  t'MU'nt. 

The  use  ol  ap[)lied  genetics  in  w heat  im|)ro\  ement 
occurs  in  close  harmony  with  total  wheat  manage- 
ment systems.  The  tarmer  must  integrate  a huge  as- 
sortiiKMit  ot  alternativ  es  in  t'ach  decision — e.g.,  an  iti- 
ili\  idual  producer  may  l)e  deciding  on  a nitrogen 
program.  It  th('  tarm  is  irrigated,  the  producer 
selects  nitrogen  amounts  and  application  timing 
based  on  soil  tests,  intended  crop  and  \arietv,  the 
end  list'  ot  that  crop,  and  watering  schedules.  II  the 
tarm  is  rainit'd,  the  product'!'  takes  into  account  soil 
tt'sts,  ci'op  considei'ations,  and  l aint'all  pi’ohahilities. 

In  hotli  cases  pi'oduct  pi'ices  at  the  time  ot  sale 
must  he  predictetl  since  they  go\ern  |)otential  gross 
returri,  w hich  in  turn  atlects  the  costs  ol  maintaining 
a (irotit  margin,  (it'iietic  inteiaction  in  this  svstem  is 
inti'icate.  The  tai'mei'  must  first  select  the  \arietv 
most  likely  to  produce  at  the  maximum  economic 
level,  for  irrigated  land,  it  may  he  a short  high- 
yielding  semidw  ai'f  either  for  the  cookie  trade  or  the 
e.xpoi't  market.  The  farmer  knows  that  part  of  the 
value  ot  his  product  is  dependent  on  low'  protein. 
However,  ina[)[)roi)riatelv  high  levels  of  nitrogen, 
w hich  greatly  improv  e yield,  will  also  raise  the  pro- 
tein of  the  crop  beyond  acce[)tahle  levels.  If  the  ex- 
port market  is  strong  and  the  total  I'.S.  supply  re- 
duced. the  higher  protein  may  he  of  little  economic 
conse(|uence. 

In  the  case  of  the  dryland  farmer,  the  variety 
selected  mav  he  taller  with  lower  yield  potential  but 
with  much  better  levels  of  adaptation  and  tolerance 
to  adverse  env  ironments.  It  may  be  designed  for  the 
bread  industry  or  the  export  market.  Part  of  the 
V alue  is  related  to  high-protein  content.  Since  mois- 
ture conservation  and  use  is  critical,  nitrogen  ap- 
plications and  amounts  must  be  selected  so  that  the 
plants  do  not  waste  their  moisture  reserve.  How  ev'er, 
nitrogen  applied  too  late  may  not  receive  enough 
rain  to  penetrate  the  soil  and  become  av  ailable  to  the 
plants.  If  the  plants  "burn  up"  because  of  unwise 
water  use  early  in  the  season,  the  seeds  will  be  high 
in  protein  but  low  in  yield.  If  inadequate  nitrogen  is 
av  ailable,  the  crop  will  generally  be  low  in  protein. 

The  abbrev  iated  protein  story  is  but  one  of  many 
examples  of  farm  management  interaction  with  ap- 
plied genetics  in  wheat  production.  Recent  changes 
in  energv'  price  and  availability,  environmental  re- 
straints, marketing  structures,  and  technology  devel- 


opment are  producing  a new  array  of  complex  prob- 
lems. 

Genetic  vulnerability  in  wheat 

Genetic  vulnerability  is  defined  as  a high  degree  of 
genetic  uniformity  in  a crop  grown  over  a wide  acre- 
age. Wheat,  which  is  produced  on  about  62  million 
acres  annually  in  the  United  States,  has  a relatively 
high  level  of  uniformity  and  genetic  v ulnuerability. 
In  1974,102  hai'd  I'ed  winter  wheat  v^arieties  were 
grown  on  36.6  million  acres,  with  four  varieties  oc- 
cupying  40  percent  ot  the  acreage.  Hard  red  spring 
wheat  varieties  totaled  80  percent  on  14.7  million 
aci'es,  with  three  varieties  occupying  52  percent  of 
the  aci'eage.  Similar  situations  occurred  with  other 
classes  ot  wheat.  Plant  pests,  including  diseases  and 
insects,  have  periodically  caused  moderate  to  severe 
vv  heuil  crop  losses  in  years  favmrable  to  the  develop- 
ment ot  strains  capable  of  attacking  current  forms  of 
resistance. 

Incoi'porating  genetic  resistance  to  pests  has  tradi- 
tionally been  the  responsibility  of  public  breeders. 
Wheat  is  a self-fertilized  plant  that  can  be  faithfully 
reproduced  from  generation  to  generation.  Private 
industry  has  been  reluctant  to  invest  R&.D  money  in 
improvements  since  the  farmer,  following  the  initial 
seed  purchase,  can  reproduce  the  crop  without  re- 
turning to  the  seed  company.  Thus,  public  breeders 
have  been  the  main  source  of  new  varieties  and  have 
had  the  responsibility  of  delivering  genetic  im- 
provements to  the  producer.  Wheat  breeding  pro- 
grams are  generally  designed  to  respond  to  State  pro- 
duction needs.  Goals  and  objectives  are  established 
by  technical  advisory  groups  that  include  breeders 
and  scientists,  growers,  use  industry  representa- 
tives, and  extension  workers. 

Genetic  variability  is  available  to  the  breeder  from 
naturally  occurring  sources  and  artificially  induced 
mutations.  Naturally  occurring  variability  bas  been 
collected  from  native  plant  populations  throughout 
the  w'orld  and  is  maintained  in  the  World  Wheat  Col- 
lection by  the  Science  and  Education  Administration 
of  USDA  located  in  Beltsville,  Md.  Currently,  about 
37,000  accessions  are  contained  in  the  collection. 
Breeders  use  the  collection  as  a reservoir  from  which 
to  draw  exotic  genes  needed  to  improve  the  value  of 
their  breeding  programs.  In  addition  to  variability 
w'ithin  w'heat  varieties,  the  breeders  can  use  special 
genetic  techniques  to  draw  valuable  genes  from  re- 
lated species  such  as  rye  and  various  forage  grasses. 
This  approach,  while  time-consuming  and  costly,  has 
been  used  in  a number  of  variety  development  pro- 
grams. Mutations  induced  by  artificial  means  have 


306  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


not  been  used  extensively  by  the  breeders,  since 
desired  mutations  without  detrimental  effects  are 
very  difficult  to  obtain.  Enough  natural  genetic 
variability  seems  to  exist  to  satisfy  needs  in  the 
foreseeable  future. 

rhe  National  Wheat  Improvement  Committee  has 
stated  that  the  World  Wheat  Collection  is  inadequate- 


ly evaluated,  characterized,  and  documented,  forc- 
ing breeders  to  spend  time  and  resources  carrying 
out  their  own  evaluation  work.  The  committee  has 
proposed  a standard  set  of  descriptors  for  all  acces- 
sions in  the  collection,  as  well  as  an  information 
management  system  to  efficiently  bring  the  informa- 
tion to  the  breeders. 


Appendix  II-B 

Genetics  and  the  Forest  Products 

Industry  Case  Study 


! 


The  Weyerhaeuser  Co. 

The  \\  everliaeuser  t\)..  u hicli  has  its  main  head- 
(luarlers  in  Centralia.  Wash,,  is  llie  largest  forest 
piodiu'ts  company  in  the  I'nited  States.  In  1970, 
Weyerhaeuser  initiated  a program  to  research  the 
! mass  propagation  of  Douglas  fir  trees  hy  tissue 
culture.  Douglas  firs  are  the  main  species  in  many  of 
the  .Nation's  forests,  o\er  S3.1  hillion  (or  about  8.5 
billion  hoard  feet)  woi  th  were  har\ested  in  1979. 
W bile  they  are  normally  [)ro[)agated  by  seed  in  the 
field,  the  classical  dexelopment  of  impro\ed  seed 
does  not  adequately  satisfy  the  criteria  of  the  rapid 
a\  ailahility  of  trees  of  superior  ciuality. 

Specially  selected  clones  ha\e  the  potential  to  dou- 
ble the  [)roducti\  ity  of  forestlands:  each  yeai'  that 
unimproved  trees  are  [)lanted  is  another  year  of 
' "suboptimum  " har\  ests  41)  years  from  now  . W ith  the 
steadily  increasing  demand  for  forest  products, 
planting  substantially  improxed  trees  as  soon  as  pos- 
sible is  of  great  economic  importance. 

V\  everhaeuser's  tissue  culture  research  began  in 
1974  w ith  a project  at  the  Institute  of  Paper  Chem- 
istry to  produce  Douglas  firs.  The  project  was  ex- 
panded w ith  a contract  for  additional  research  at  the 
Oregon  Graduate  Center.  .Although  the  intention  w as 
! to  propagate  select  strains  of  mature  trees,  the  main 
focus  of  the  program,  in  1974  to  1978,  was  to  de\el- 
op  a basic,  consistent  system  for  propagation.  From 
1978  to  the  present,  Weyerhaeuser  has  been  con- 
I ducting  most  of  its  applied  research  into  Douglas  firs 
’ at  its  ow  n research  facilities  in  Centralia,  Wash.  Basic 
research  is  still  being  funded  at  the  Institute  of  Paper 
j Chemistry,  which  serx  ices  the  entire  forest  industry. 
W hile  specific  figures  for  the  tissue  culture  systems 
research  haxe  not  been  made  available,  the  annual 
research  and  development  budget  at  Weyerhaeuser 
specifically  for  biological  xvork  xx  ith  forest  species  is 
on  the  order  of  S7  million  to  S8  million.* 

The  project  in  mass  propagation  of  Douglas  fir  by 
tissue  culture  xxas  initiated  to  establish  a reliable, 
economic  means  for  mass  production  of  superior 
trees.  The  cloning  of  these  trees  could  bring  higher 


'Rex  XIcCulloiigh.  The  W eyerhaeuser  Co.,  personal  communication  (.Xlay 
1980)  with  the  Plant  Resources  Institute  in  the  working  report.  Commercial 
L'ses  of  Plant  Tissue  Culture  and  Potential  Impact  of  Genetic  Engineering  on 
Forestry,  prepared  under  contract  to  O I'  X,  1980. 


yields  and  shorter  harvest  cycles,  as  well  as  rapid 
production  of  tree  stands  for  seed  production. 

The  immediate  results  of  10  years  of  research  are 
not  overly  impressive  at  first  glance.  To  date,  3,000 
tissue-cultured  Douglas  firs  have  been  planted  for 
comparison  analysis  and  research  of  handling  tech- 
niciues,  transfer  procedures,  etc. 

The  cost  effectiveness  of  a tissue  culture  program 
is  determined  by  several  factors,  of  which  labor  in- 
tensity xaries  the  most.  The  more  streamlined  the 
system  can  be  made,  the  fewer  labor-requiring  steps 
that  are  needed— the  less  direct  costs  will  be  in- 
curred. Ideally,  cells  xvould  be  cultured  in  sterile  con- 
ditions and  then  planted  for  the  direct  embryogene- 
sis  of  plantlets  that  are  ready  for  the  field.  Steps  that 
inx  olx  e cutting  shoots  and  rooting  them  on  another 
media  or  repeated  subculturing  procedures  are  cost- 
ly and  cumbersome.  The  major  problem  affecting 
cost  so  far  is  the  difficulty  of  achieving  high  volume 
plant  regeneration  from  the  tissue  cultures.  Efficient 
systems  xvith  more  successful  regeneration  will  re- 
duce the  labor  and  materials  involved  in  culturing 
and  result  ultimately  in  a lower  cost  per  plant. 

In  addition  to  problems  of  cost,  Weyerhaeuser  has 
run  into  the  classic  difficulty  with  woody  species— 
the  inability  to  obtain  required  results  from  plants 
more  than  1 year  old.  In  addition,  the  risk  of  induced 
genetic  variability  increases  wdth  every  subculture  of 
the  tissues.  The  triggering  techniques  for  effective 
manipulation  of  mature  versus  embryonic  and  imma- 
ture tree  tissues  are  not  well  understood,  and  un- 
locking the  Douglas  fir  system  may  well  provide  in- 
sight into  some  basic  physiological  questions. 

Some  commercial  companies  do  not  want  to  get 
deeply  involved  in  basic  research  because  it  is  ex- 
tremely expensive  and  time-consuming.  However,  it 
has  been  up  to  the  major  forestry  companies,  such  as 
Weyerhaeuser,  to  independently  fund  essentially 
basic  research  into  the  biological  triggers  for  organo- 
genesis and  embryogenesis  of  Douglas  fir. 

By  comparison,  no  other  plant  bas  been  as  intense- 
ly researched  for  mass  propagation  purposes  and 
proved  so  unyielding.  Among  other  things,  this  in- 
dicates that  questions  of  basic  plant  cell  physiology 
xvill  have  to  be  addressed  before  major  break- 
throughs can  be  expected.  The  goals  of  the  Weyer- 
haeuser program  are  exacting  and  demand  the  re- 
finement of  present  techniques  into  a precise  in- 


307 


I; 


308  • Impacts  of  Applied  Genetics — Micro-Organisms,  Piants,  and  Animals 


dustrial  science.  While  it  may  seem  that  the  invest- 
ment has  been  disproportional  to  the  returns  at  this 
point,  it  must  be  remembered  that  they  are  the  fore- 
runners of  a new  technology,  both  in  terms  of  work- 
ing with  mature  tree  tissues  of  an  especially  intricate 
species  and  in  terms  of  imposing  stringent  industrial 
standards  on  a mass  biological  production  system. 

Simpson  Timber  Co. 

The  Simpson  Timber  Co.,  whose  central  headquar- 
ters are  in  Seattle,  Wash.,  is  a large  producer  of  red- 
wood and  other  forest  products,  and  has  been  in- 
volved over  the  past  5 years  in  a program  to  develop 
a mass  production  system  for  the  coast  redwoods 
through  tissue  culture.  Approximately  $250,000  has 
been  invested  in  research  performed  at  the  Universi- 
ty of  California,  Irvine,  by  Dr.  Ernest  Ball,  a recog- 
nized authority  in  the  field  of  tissue-cultured  red- 
woods.^ 

Coastal  redwoods  are  normally  a field-seeded  crop 
and  have  a production  cycle  of  around  50  years.  The 
major  reason  for  consideration  of  tissue  culture  over 
seed  is  the  greater  speed  with  which  superior  trees 
might  be  developed  through  tissue  culture  as  com- 
pared to  using  seed  stock.  Simpson  Timber  Co., 
which  has  been  involved  in  a controlled  breeding 
program  along  conventional  lines  as  well,  and  is  ap- 
proaching the  creation  of  homozygous  strains.  Since 
a sequoia  seedling  does  not  reach  sexual  maturity 
before  it  is  15  to  20  years  old,  and  since  about  10 
generations  are  normally  required  to  produce  a ti'ue 
homozygous  strain,®  the  classical  process  is  time-con- 
suming and  contains  no  guarantees  that  the  end 
products  will  he  better  than  the  clones  selected 
through  tissue  culture. 


^I'.rncsl  liiil!,  UnivtM'sily  of  (ialitbrnia,  Irvine,  pcM'sonal  roiniminicalion 
{May  n)80)  with  the  Plant  Resources  Inslituli’  in  iIk*  working  ri’port.  ('om- 
inrrcinl  [ 'scs  of  iHiinl  I'issuv  ( ullurr  ninl  Tolrnlinl  Impiirl  ofCriirlir  l.nj;,inrrr- 
//It'  0/1  l'(H'(‘slry,  prepart'd  under  contract  to  ()  1 \. 

'.lames  Radelius.  Simpson  l imher  Co.,  personal  ('omiminicalion  (May 
with  the  Plant  Resourc(*s  Inslilule  in  the  wtirkin^  rep(»ri,  ( Dtwnrrrinl 
LLsrs  of  l*lnnl  I'issin'  (Uilliirc  and  Tolrntinl  impnet  of  (',(‘nrlir  on 

l^'orrslrv.  prepared  under  conlract  to  ()  I \, 


Elite  trees  are  selected  from  wild  stands  for 
straightness  of  trunks,  height.  s|)ecific  gravity  of 
wood,  and  proper  branch  drop  (bl  anches  that  drop 
without  tearing  the  stem).  I'here  are  no  major  pests 
in  redwoods,  so  pest  and  disease  resistance  have  not 
been  a concern.  Two  methods  of  si'lection  are  used. 
Clones  of  special  trees  are  pioduced  by  rooting  the 
uppermost  branches  of  the  tree,  a process  (hat  takes 
up  to  1 year.  .Although  the  rooting  percentage  may 
he  as  low  as  10  percent,  this  method  has  the  advan- 
tage of  producing  mature  i loned  plants  that  can  I'on- 
tinue  to  |)roduce  flow  ers  and  .seed  Simpson  is  using 
roughly  200  elite  trees  for  these  clones 

Elite  trees  can  also  prov  idi’  clones  througli  tissue 
cultures  of  their  nei'dies,  a process  that  is  less  time- 
consuming  hut  which  |)roduces  seed  veiv  slowly 
because  of  the  time  involved  in  maturation  Simp.son 
rimher  Co.  has  planted  out  2.500  tissue  cultured  red- 
woods for  fii'ld  comparisons  with  seedling  material. 
The  results  so  far  have  heiMi  encouraging,  luit  it  may 
take  another  10  to  15  years  helore  delmite  conclu- 
sions can  l)('  draw  n I hi'  in.ijor  (actors  hiMug  .ma- 
lyzed  arc’  fic’ld  grow  th  rates  and  oulplantmg  surv  iv  .il 
percentages.  ( lones  of  I’lite  v .irietic’s  w ill  also  liav c-  to 
he  compared  to  the  p.irent  trees  lor  the  trails  origi- 
nally sele.cled  for.  such  as  wood  (|u.ililv  Mnc c the 
opc’iational  cost  ot  tissue-cultured  planllels  is  about 
twice  that  ol  seedlings,  the  c|ualilv  ol  tissue  c iillured 
plants  must  he  niarkediv  superior  it  the  program  is 
to  he  cost  c’flectiv  e 

Dr.  Ball  is  conlideni  that  the  tissue  c ullure  sv  stem 
w hich  has  been  di’v c’lopi’d  lor  the  r.ipid  mulliplua- 
lion  of  elite’  rc’dwood  trees  is  readv  lor  nnpli-menla- 
lion  at  a commercial  produi  lion  lac  ililv  * 'Minpson 
I imhc’i'  ( o.  is  planning  the  c ciosli  ui  lion  ol  ,i  lissup 
culture  lah  at  their  ( alilorma  head(|uaiicrs  within 
ihc’  next  2 yc’ars  I he  pilot  plant  is  rxpi  c led  to  c iiM 
$250. ()()()  and  produc c upwards  ol  200000  planllels 
in  its  lirsi  vear  ol  produi  lion  \s  ni.iss  priHlui  lion 
Ic’chniciuc’s  are  perlec  led  Ihi-  i iimpanv  pl.nis  to  ex- 
pand the  lacililv  to  ,i  produi  tion  capai  ilv  ot  over  I 
million  planllels  per  v ear  ' 

'Ikill.  n|i  I II 

'R.iilriiiis  np  « il 


Appendix  II-C 

Animal  Fertilization  Technologies 


Sfterm  stnrufiv 

1)1  I IM  I ION 

I  hr  Irtf/ui^  ol  to  — 196®  I >lora#{e  tor  an 

iiuit'tinilr  limr  toliov\t‘d  hv  tha\\mf<  aiul  .successtul 
inM'iiunalion 

sr\n Ol  no;  \iu 

t'oiurplion  rali'^  at  fii  r»t  insrmination  v\itlt  tro/en 
sfMTin  a\rra>{t*  )M-tut‘<>n  'll)  to  6a  |HTifnt  tor  most 
sfMTirs  This  t»t  finok»^>  is  not  a krv  to  the  siurrss  ot 
artitirial  iiiM'inindtion  lAII.  hut  tMt-aiisr  ot  thr  con- 
vrniriur  it  is  now  an  rss»*ntial  iiiftriMilrnt  (airmit 
o|M‘rational  priM  rtiuri's  an*  a(ii*<|uatt'  tor  thi*  dairy  in- 
dustry 

\i)\  \M'\r;Fs 

1 (.rratiT  use  ot  seUt  tnl  tnills  as  \I  studs 

2 FJimination  ot  the  netti  to  maintain  e.\fM*nsi\i*  and 
danjjerous  hulls  on  dair>  farms 

3 Sf)«*rm  ran  lx*  testetl  for  diseas«*  arul  treated  for 
venereally  transmittetl  diseases 

4 Fase  ot  transfxirt  anti  theit’fore  of  increasing?  po- 
tential offspring 

n I I hf; 

Little  change  is  anticipated  in  semen  processing. 
Freeze-druxl  semen  is  unlikely  to  be  successful 
enough  to  use  Sperm  banking  can  be  e.\|>ected  to  in- 
crease. especially  on  .-\l  studs.  Banking  provides 
cheap  storage  while  bulls  (slaughtered)  are  being 
progeny  tested,  and  insurance  against  loss  of  bulls 
through  natural  causes.  For  preserAation  of  semen 
from  bulls  of  less  populous  breeds,  banking  can  be 
completed  in  about  a year  after  which  the  bull  can 
be  slaughtered. 

Artificial  insemination 

DEFIMTIO.V 

Manual  placing  of  sperm  into  the  uterus. 

STATE  OF  THE  ART 

Highly  developed  for  most  spiecies.  Representative 
use  rates  in  the  United  States  are:  dairy  cattle,  60  per- 
cent: beef  cattle,  5 percent:  turkeys,  100  percent. 
The  major  limitation  to  the  use  of  .AI  is  the  low  na- 
tional average  conception  rate  at  first  service, 
around  50  percent.  The  success  or  failure  of  ,AI  is 
determined  by  a multiplicity  of  factors  including 


estrus  detection,  i]uality  of  semen,  timing  of  in- 
semination, and  semen  handling. 

DISADVANTAGES 

1.  U ith  increased  herd  size,  estrus  detection  has 
become  a major  problem. 

2.  Ine.vperienced  dairymen  are  buying  semen  and  in- 
seminating (heir  own  cows,  resulting  in  lowered 
fertility  and  no  feedback  on  semen  fertility. 

ADVANTAGES 

1 Widespread  use  of  genetically  superior  sires. 

2.  St*rvices  of  jiroven  sires  at  a lower  cost. 

3.  Klimination  of  cost  and  danger  of  keeping  bulls  on 
the  farm. 

4 Control  of  certain  diseases. 

5.  Use  of  other  bretnling  techniciues  including  cross- 
breeding. 

6.  ('ontinued  use  of  valuable  sire  after  his  death. 

FCTCRE 

Greater  use  of  AI  in  beef  cattle  will  depend  on  the 
availability  of  successful  and  inexpensive  estrus  syn- 
chronization technology',  on  relaxed  restrictions  of 
the  various  breed  associations,  and  on  accurate  prog- 
eny records. 

Estrus  synchronization 

DEFINITION 

Estrus  ("heat"),  is  the  period  during  which  the 
female  will  allow  the  male  to  mate  her.  This  sexual 
behavior  is  subtle  and  varies  widely  among  individ- 
uals. Thus  the  synchronization  of  estrus  in  a herd, 
using  various  drug  treatmnts,  greatly  enhances  AI 
and  other  reproduction  programs. 

STATE  OF  THE  ART 

Effective  methods  for  synchronization  of  estrus 
periods  for  large  numbers  of  animals  have  been 
available  for  more  than  two  decades,  and  several  ap- 
proaches are  now  available  which  result  in  normal 
fertility.  Several  schemes  involve  use  of  prosta- 
glandin Fi  (PGFz)  for  the  cow  and  ewe.  However,  FDA 
approves  usage  only  for  controlled  breeding  in  beef 
cows  and  heifers,  nonlactating  dairy  heifers,  and  in 
mares. 

ADVANTAGES 

1.  Time  a heifer’s  entry  into  a milking  stream. 

2.  Increase  productivity  by  breeding  heifers  earlier 
in  life. 


309 


310  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animais 


3.  Ability  to  breed  large  numbers  of  cattle  over  a 
shorter  calving  interval. 

4.  Increase  use  of  Ab  expecially  in  beef  cattle,  sheep, 
and  swine. 

FUTURE 

Estrus  cycle  regulation  should  allow  selected  sires 
to  be  more  widely  used  to  improve  important  traits 
in  beef  cattle.  It  should  also  gain  widespread  and 
rapid  acceptance  among  dairymen  as  well. 

Superovulation 

DEFIMTION 

Superovulation  is  the  hormonal  stimulation  of 
multiple  ovarian  follicles  resulting  in  release  from 
the  ovary  of  a larger  number  of  oocytes  (ova)  than 
normal. 

STATE  OF  THE  ART 

Superovulation  with  implantation  into  surrogate 
mothers  increases  the  number  of  offspring,  usually 
from  highly  selected  dams.  Adequate  procedures  are 
presently  available  for  superovulation  of  laboratory 
and  domestic  animal  species,  except  the  horse.  The 
drugs  used  to  induce  superovulation  are  the  go- 
nadotropins, pregnant  mare’s  serum  gonadotropin 
(PMSG)  and  follicle  stimulating  hormone  (FSH),  in 
some  instances  followed  by  other  treatments  to  stim- 
ulate ovum  maturation  and  ovulation.  Superovulated 
ova  result  in  normal  offspring  with  the  same  success 
rates  as  achieved  with  normally  ovulated  ova. 

DISADVANTAGES 

1.  Greatest  drawback  is  that  degree  of  success  can- 
not be  predicted  for  an  individual  animal. 

2.  Batches  of  hormones  for  ovulation  treatment  vary 
widely  in  quality. 

3.  PMSG  is  scarce,  and  has  been  declared  a drug  by 
the  Food  and  Drug  Administration  (FDA).  Thus, 
most  use  of  PMSG  is  now  illegal. 

4.  There  is  insufficient  data  to  judge  the  effect  of 
repeated  superovulation. 

FUTURE 

Methods  for  superovulation  will  improve  consist- 
ency of  results.  Additional  understanding  of  basic 
physiological  mechanisms  will  facilitate  such  efforts. 
New  work  in  superovulatory  technology  involves  ac- 
tive immunization  against  adrostenedione  (a  hor- 
mone involved  in  regulation  of  follicular  develop- 
ment). This  treatment  prevents  atresia  and  reliably 
increases  the  frequency  of  multiple  ovulations.  The 
technology  has  definite  commercial  potential  for  cat- 
tle husbandry  and  limited  potential  for  sheep  hus- 


bandry, and  much  current  effort  is  directed  towards 
developing  and  testing  a commercial  procedurtv 

Embryo  recovery 

DEFINITION 

The  collection  of  the  fertilized  o\a  from  the 
oviducts  or  uteri.  Collection  of  embryos  is  a 
necessarv  step  for  embryo  transfer  or  storage,  and 
for  many  experiments  in  reproductixe  hiologx’  Both 
surgical  and  nonsurgical  methods  are  used. 

STATE  OF  THE  ART 

Surgical.— Methods  are  axailahU*  for  recowring 
40  to  80  percent  of  oxulations  from  cattle,  slu-ej). 
goats,  swine,  and  horses.  Fhe  d(*\rlopment  of  adhe- 
sions and  scar  tissue  following  surgery  limits  these 
techniques.  Surgical  recovery  is  th(‘  only  method  lor 
sheep,  goats,  and  pigs.  It  is  pre.senlly  practiicd 
almost  exclusively  when  a suspecti'd  |)alh()logv  ol  the 
oviducts  renders  an  individual  suhl(*rtile.  or  when 
emhrvos  must  he  recovered  hefon*  the  individual 
reaches  puberty. 

Nonsurgical.— Non-surgical  embryo  n‘(ov  erv 
technitiues  are  preferred  for  the  cow  .iiul  horse 
Fiftv  to  eightv  jtercent  of  cow  ovulations  lan  h<* 
recovered,  and  40  to  00  percent  ol  the  operations  on 
horses  to  recover  the  single  ov  illation  are  suci  esslul 

AD\  AN  r \(;i;s 

1.  Nonsurgical  (>mhrvo  transler  can  he  perlormed  .m 
unlimited  number  of  times 

2.  Requirements  for  eiiuipment  |)ersonnel  ami  time 
are  low  in  nonsurgical  recovery  I his  is  espei  lallv 
important  in  milk  cattle:  since  the  nonsui  gii  al  jiro 
cedure  is  performi'd  on  the  larm  milk  prodm  tion 
is  not  interrupted 

3.  A single  embryo  can  he  obtained  between  super 
ovulation  treatmcMits 

4.  Emhrvos  can  he  obtained  Irom  <i  voung  heiler 
before  it  reaches  puberty 

5.  The  technologv  is  especiallv  import.mt  lor  re 
search,  e g.,  in  (‘fforls  to  |)roduce  idenlii  al  twins 
embryo  biopsies  for  se\  determin.ition  eti 

lilt  Rl 

Methods  of  collecting  emhrvos  have  not  i h.ingeil 
appreciably  since  about  I!l7li  noi  are  sigmlu  ant  ad 
vances  predicted  for  the  luture 

Embryo  transfer 

DI.I  IM  I lOV 

Implantation  of  an  emhrvo  into  lln  ovnlmt  m 
uterus. 


Appendix  ll-C— Animal  Fertilization  Technologies:  *311 


SI  \ll  ()l  IMh  \HT 

Siirijiriil.  I’rf^iuiiu  \ ralrs  ol  51)  to  75  [WiTent 
art-  arhi«‘\  ahlf  m fov\>  sfut*[),  goats,  pigs,  and 
hoiM'-*  Surgu  al  transtcr  is  tin*  onlv  piMi  tical  nu*tho<i 
in  shi*fp  goals  anil  jiigs  and  is  lh«*  pivdoininant 
in«‘thod  tor  » ov\  s and  hoi  srs  \ nuin()(*r  of  factors 
d»‘t»*rmini‘  itu*  smiTss  ot  surgical  transfer:  age  and 
(|u.ilit\  of  eiiif)r\os  >iti*  of  transfer,  ift*grt*t*  of  s\n- 
chroiu  fH'lween  eNlroiis  cycles  of  the  donor  and  re- 
(ipients  nuinfier  of  emfiryos  transferred  in  \itro 
culture  conditions  skill  ot  jM’rsoniiel.  and  manage- 
m»*nt  l**cfmi(|ues  I tie  50-  to  HO  jiercent  success  rate 
in  cattle  comjiarf's  witfi  M sin  cess  rales  at  first  ser\  • 
iie  (I’regnam  v rales  stiould  not  fie  confused  with 
suri  \\ al  rates  hich  mav  fie  mucfi  low  er  I 

Nonsiirgicul.  - I his  method  is  an  adaptation  of 
\l  Rejnirted  succevs  rates  are  much  lower  tfian 
those  wilfi  surgical  transfer  .\onsurgical  transfer  is 
not  useil  in  sfi*t*p  go.its  or  pigs 

\l)\  AN  I \(;i  s 

1 Ofitainmg  offspring  fmm  females  unalile  to  su[i- 
(Mirt  pregnanc\ 

2 Ofitaming  more  offspring  from  valuafile  females. 

3 U ilh  a homo/.ygous  donor  undesirahle  riH-essii  e 
traits  among  animals  used  for  \l  can  he  rapidiv 
detivteil 

4 Introducing  new  genes  into  s}M*cific  pathogen-free 
sw  me  herds 

5.  foupletl  with  short-  or  long  term  emhrvo  storage. 

trans()ortation  of  animals  as  emfir\(is 
H Increasing  the  (lopulation  fiase  of  rare  or  endan- 
geretl  breeds  of  animals  hv  use  of  closely  related 
breeds  for  ret'ipienls 

7.  Separation  of  embryonic  and  maternal  influences 
in  research 

ms  \n\  \ NT  AGES 

f.  Personnel  requirements  in  surgical  transfer  ac- 
count for  a large  share  of  high  costs  and  thus  limit 
applicability  in  animal  agriculture. 

2.  Prm  ision  of  suitable  recipients  is  the  greatest 
single  cost  in  embryo  transfer. 

FI  Tl  RE 

Surgical  transfers  will  remain  the  method  of 
choice  for  sheep  goats,  and  pigs  in  the  foreseeable 
future.  For  cows  and  horses,  however,  nonsurgical 
methods  will  be  increasingly  used  rather  than  sur- 
gical techniques  (and  this  will  be  apparent)  within 
the  ne.xt  year  or  two.  It  is  likely  that  half  of  the  com- 
mercial transfer  pregnancies  in  cattle  in  North  .Amer- 
ica in  1980  v\ill  be  done  nonsurgically,  even  if  suc- 
cess rates  are  only  60  to  80  percent  of  those  obtain- 
able with  surgical  transfer.  .Among  future  appli- 
cations. a role  for  embryo  transfer  can  be  predicted 


in  progeny  testing  of  females,  obtaining  twins  in  beef 
cows,  obtaining  jirogeny  from  prepubertal  females, 
and  in  combination  with  in  \'itro  fertilization  and  a 
\ariety  of  manipulative  treatments  (e.g.,  production 
of  identical  tw  ins,  selling,  genetic  engineering,  etc.) 

Embryo  storage 

DEFINITION 

•Maintenance  ot  embryos  for  several  hours  or  days 
(short-term)  or  for  an  indefinite  length  of  time  (freez- 
ing). 

STATE  OF  THE  ART 

Short-U?rni.— The  requirement  for  embryos 
from  farm  animal  species  has  not  been  defined, 
although  adequate  culture  systems  for  the  short  in- 
ter\al  between  recovery  and  transfer  have  been 
developed  by  trial  and  error.  Whereas  the  important 
parameters  of  culture  systems  have  been  identified 
(e  g.,  temperature,  pll,  etc.),  optimal  conditions  have 
not  been  determined.  Cow  embryos  may  be  stored 
for  three  days  in  the  ligated  oviduct  of  the  rabbit. 

Long-lemi  (I'rtiezing).— No  completely  adequate 
protocol  e.xists  for  freezing  embryos  of  farm  species. 
One-third  to  two-thirds  of  embryos  are  killed  using 
present  methods.  Pregnancy  rates  of  32  to  50  per- 
cent for  cattle,  sheep,  and  goats  have  been  reported 
after  freezing.  No  successful  freezing  of  swine  or 
horse  embryos  followed  by  development  to  term  has 
been  reported.  Despite  disadvantages  (one-half  of 
embryos  are  often  killed)  advantages  are  such  that  in 
some  situations  embryo  freezing,  and  embryo  sell- 
ing. are  already  profitable. 

ADVANTAGES 

1.  .Amplification  of  advantages  of  embryo  transfer. 

2.  Elimination  of  requirements  for  large  recipient 
herds  when  embryo  transfer  is  being  used. 

3.  Reduction  of  costs  in  animal  transport. 

4.  Control  of  genetic  drift  in  animals  over  prolonged 
time  interx’als. 

FUTURE 

Anticipated  development  of  embryo  culture  tech- 
nology w'ould  be  of  significance  in  efforts  toward  in 
\'itro  maturation  of  gametes,  in  vitro  fertilization,  sex 
determination,  cloning,  and  genetic  engineering,  all 
of  which  involve  prolonged  manipulation  of  gametes 
and  embryos  outside  of  the  reproductive  tract. 

As  freezing  rates  improve,  nearly  all  embryos  re- 
covered from  cattle  in  North  America  will  be  frozen. 
Probably  as  many  as  half  of  the  embryos  will  be 
deep-frozen  for  2 to  3 years.  It  is  unlikely  that  suc- 
cess rates  will  ever  approach  90  percent  of  those 


312  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


without  freezing.  However,  70-  to  80-percent  success 
rates  may  be  attainable  within  several  years.  It  ap- 
pears that  embryos  can  be  stored  indefinitely  with 
little  deterioration. 

Sex  selection 

DEFINITION 

Tests  to  determine  the  sex  of  the  unborn  or  deter- 
mination of  sex  at  fertilization  by  separating  x-  bear- 
ing from  y-bearing  sperm. 

STATE  OF  THE  ART 

Sexing  of  embryos.— Through  karyotyping 
nearly  two-thirds  of  embryos  can  be  sexed.  Tech- 
niques using  identification  of  the  condensed  X chro- 
mosomes are  unreliable.  A third  method,  identifica- 
tion of  sex-specific  gene  products,  is  under  develop- 
ment. 

Sexing  of  sperm.— A 100-percent  method  has 
not  been  achieved  in  any  mammalian  species;  and  no 
standard  protocol  for  farm  species  exists. 

FllTlIRE 

Before  this  technology  can  be  applied  commercial- 
ly, it  must  be  simple,  fast,  inexpensive,  reliable,  and 
nonharmful  for  embryos.  Such  techniques  could  un- 
doubtedly be  developed.  There  would  be  numerous 
medical  and  experimental  applications. 

There  is  much  interest  in  research  in  this  area 
because  of  its  use  in  understanding  male  fertility 
with  AI  in  bumans,  and  in  enhancing  sperm  survival 
after  frozen  storage. 

Twinning 

DEFINITION 

Artificial  production  of  twins,  either  using  embryo 
transfer  or  hormone  treatments. 

STATE  OF  THE  ART 

Currently,  embryo  transfer  is  tbe  most  effective 
method  for  inducing  twin  pregnancies  in  cattle, 
resulting  in  pregnancy  rates  of  between  67  to  91  per- 
cent, of  which  27  to  75  percent  deliver  twins.  Other 
methods  include  transferring  one  embryo  into  a cow 
which  has  been  artificially  inseminated,  and  hor- 
monal induction  of  twinning,  which  is  a modification 
of  superovulation.  This  latter  method  is  not  reliable. 

ADVANTAGE 

The  advantage  of  twinning  in  nonlitter-bearing 
species  is  the  improved  feed  conversion  ratio  of  pro- 
ducing the  extra  offspring. 


DISADVANTAGE 

The  major  disadvantage  of  twinning  is  intensive 
management  necessary  for  periparturient  conqtlica- 
tions,  unpredictable  gestation  periods,  depressed  lac- 
tation, etc. 

FUTliRE 

Technical  feasibility  for  twinning,  in  conjunction 
with  embryo  transfer,  management  adjustments, 
and  selection  for  good  recipients,  can  be  predicted.  \ 
reliable  procedure  for  twinning  in  sheep  can  alst)  be 
expected.  The  technologv'  would  most  likely  be  first 
used  in  Europe  and  Japan,  whei  e there  are  shortage's 
of  calves  to  fatten  for  beef. 

In  vitro  fertiliz.ation 

DEFINITION 

rhe  union  of  egg  and  sperm  outside  the  re[)i  ()du( - 
tive  tract.  For  some  species,  the  technology  includes 
successful  developmetit  of  the  embryo  to  gestation 
and  birth. 

STATE  OF  'HIE  AR  I' 

In  vitro  fertilization  has  been  accomplished  in 
several  laboratory  animal  sjiecies,  including  the  rab- 
bit, mouse,  rat,  hamster,  and  guinea  pig  and  nine 
other  mammalian  nonlaboratorv  species,  including 
man,  cat,  dog,  pig,  shee|),  and  cow  lloucver,  norm.il 
development  following  in  \ilro  tertili/alion  and  cm 
bryo  transfer  has  only  been  accom|)lished  in  the  rab 
bit,  mouse,  rat,  and  human  ( onsisteni  .md  lepe.il 
able  success  with  in  \ itro  lertili/.alion  in  larm  spei  les 
has  not  yet  been  accomplished 

None  of  th(?  ca.ses  of  reported  siici  ess  ot  in  \ilm 
fertilization,  embryo  Iranster.  and  normal  de\i*lop 
ment  in  man  is  well  documented 

Most  of  the  in  vitro  lerlili/.alion  work  to  d.ile  has 
concentrated  on  the  development  ol  a leseaich  tool 
so  that  the  physiological  and  bioi  hemical  events  m 
fertilization  and  early  devi-lopment  could  be  In-tter 
understood.  More  practic.il  .ipplii  .ition  ol  m vitro 
fertilization  techni(|ues  would  iiu  lode 

1. a  means  for  as.sessing  the  lerlililv  o!  ovum 
and/or  sperm: 

2.  a means  to  overcome  lem.ile  inlet  tilitv  w ilh  em 
bryo  transfer  into  .i  recipient  .mim.il  .ind 

;i.  when  coupled  with  ovum  .md  or  embrvo  situ 
age  and  transfer.  .i  me.ins  to  l.i<  ilit.ile  t ombm.i 
tion  of  selected  ov  a w ilh  selet  ted  sperm  loi  pm 
duction  of  indiv  iduals  w ilb  iiredicled  < h.ii  ,ii  lei 
istics  at  an  appropri.ile  time 


Appendix  II  C— Animal  Fertilization  Technologies  • 313 


n n HK 

Kapiil  progrt's.s  in  restMirh  is  anticipated  and 
main  of  th»*  (xitential  applications  of  in  \ itro  fertiliza- 
tion to  animal  breeding  should  become  practical 
VMthin  the  next  ft)  to  20  years  V\  ith  furtber  develop- 
ment of  in  vitrt)  fertilization  methoilolo^v . along  with 
storage  of  unfertilizetl  (kh  v fes  (gamete  banking),  fer- 
tilization of  desiretl  crosses  slunilil  become  possible. 
In  the  more  distant  fiiturtv  genetic  engineering  and 
sjH*rm  >e\ing  along  with  in  vitro  fertilization  may 
iHH  ome  fxissible 

t*itrtherutfivncsis 

i)t:i  IM  noN 

The  initiation  of  dev(>lopmi*nt  in  the  absense  of 
sjx'rm 

STMKOI niK  \HT 

Parthenogenesis  has  not  been  satisfactorily  dem- 
onstrattxl  or  describetl  for  mammalian  species.  The 
lx*st  available  information  leads  to  the  conclusion 
that  maintenance  of  parthenogenetic  development  to 
prixluce  normal  offspring  in  mammals  approximates 
imfxissibilitv 

(Inning:  pradiictian  of  identical  tit  ins 

nUFIMTIOV 

The  protiuction.  using  a variety  of  methods,  of 
genetically  identical  indiv  iduals. 

ST  \TE  OF  THE  ART 

There  are  several  ways  to  obtain  genetically  iden- 
tical livestock.  The  natural  way  is  identical  twins, 
although  these  are  rare  in  sp€*cies  other  than  cattle 
and  primates.  Both  natural  and  laboratory  methods 
depiend  on  the  fact  that  the  blastomeres  of  early  em- 
bryos are  totipotent  (i.e..  each  cell  can  develop  into  a 
complete  individual  if  separated  from  the  others.) 
For  practical  purposes,  highly  inbred  lines  of  some 
mammals  are  already  considered  genetically  iden- 
tical: F,  crosses  of  these  lines  are  also  considered 
genetically  identical  and  do  not  suffer  from  the 
depressiv  e effect  of  inbreeding. 

AD\  A.VTAGE 

•An  advantage  of  identical  twins  is  the  e.xperimen- 
tal  control  provided  by  one  animal  through  which 
two  sets  of  environmental  conditions  can  be  com- 
pared for  effects  on  certain  end  points,  e.g.,  native  v. 
surrogate  uterine  environments  for  gestational  de- 
velopment, nutrition  on  milk  production,  etc. 


Cloning:  nuclear  transplantation 

DEFIMTION 

The  production  of  genetically  identical  mammals 
by  inserting  the  nucleus  ot  one  cell  into  another, 
before  or  after  destroying  the  original  genetic  com- 
plement. These  occur  by  separation  of  embryos  or 
parts  ot  embryos  early  in  development  but  well  after 
fertilization  has  occurred. 

STATE  OF  THE  ART 

h.xpeM'imentalists  have  found  in  certain  amphibia 
that  transplantation  of  a nucleus  from  a body  cell  of 
an  embryonic  (tadpole)  stage  into  a zygote  following 
destruction  or  removal  of  the  normal  nucleus  can 
lead  to  development  of  a se.xually  mature  frog. 

FllTDRE 

The  ideal  technique  for  making  genetic  copies  of 
any  giv'en  outstanding  adult  mammal  would  inv^olve 
inserting  somatic  (body)  cell  nuclei  into  ova,  which 
may  take  years  of  work  to  perfect  if  indeed  it  is  possi- 
ble. There  is  some  evidence  that  adult  body  cells  are 
irreversibly  differentiated. 

How  identical  will  clones  be?  They  can  be  ex- 
pected to  be  fairly  similar  in  appearance.  They  would 
be  less  similar  than  identical  twins,  however,  which 
share  ooplasm  and  uterine  and  neonatal  environ- 
ments. Furthermore,  certain  components  are  inher- 
ited exclusively  from  the  mother,  e.g.,  the  mitochon- 
drial genome  and  perhaps  the  genome  of  centrioles. 
The  random  inactivation  of  one  or  the  other  of  the  X 
chromosomes  may  also  limit  similarities.  Other  dif- 
ferences among  clones  would  result  from  the  pre- 
natal environment;  in  litter-bearing  species  even 
uterine  position  can  affect  offspring.  In  single-bear- 
ing species  the  maternal  effect  may  be  pronounced. 
Environmental  differences  in  later  life  may  greatly 
affect  certain  traits,  even  if  those  traits  have  a strong 
genetic  component. 

Serious  technical  barriers  must  be  overcome 
before  realistic  speculation  of  possible  advantages  in 
animal  production  can  be  foreseen. 

Cell  fusion 

DEFINITION 

The  fusion  of  two  mature  sex  cells  or  the  fertiliza- 
tion of  one  ovum  with  another.  An  analogous  scheme 
for  the  male  would  be  accomplished  by  microsurgi- 
cal  remov'al  of  the  female  pronucleus  and  substitu- 
tion of  nuclei  from  two  sperm.  Combining  sex  cells 
from  the  same  animal  is  called  “selfing.” 


314  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


STATE  OF  THE  ART 

Combination  of  ova  has  led  to  early  development 
to  the  blastocyst  stage  in  the  mouse  but  no  further 
development  following  transfer  has  been  reported. 
Initial  success  in  experimentation  with  manipulation 
of  pronuclei  has  been  reported. 

FUTURE 

Cell  fusion  technology  may  someday  prove  useful 
for  getting  genetic  material  from  a somatic  cell  into  a 
fertilized  1-cell  embryo  for  the  purpose  of  cloning.  In 
conjunction  with  tissue  culture  technology  the  tech- 
nology would  have  a role  in  gene  mapping  of  chro- 
mosomes for  the  cow  and  perhaps  other  species. 

Combining  ova  of  the  same  animal,  "selfing,” 
would  rapidly  result  in  pure  genetic  (inbred)  lines  for 
use  as  breeding  stocks.  The  technique  would  also 
lead  to  rapid  identification  of  undesirable  recessive 
traits  which  could  be  eliminated  from  the  species. 

Chimeras 

DEFINITION 

A chimera  is  an  animal  comprised  of  cell  lines 
from  a variety  of  sources.  They  can  be  formed  by 
fusing  two  or  more  early  embryos  or  by  adding  extra 
cells  to  blastocysts. 

STATE  OF  THE  ART 

Live  chimeras  between  two  species  of  mouse  have 
been  produced.  Such  young  have  four  parents  in- 
stead of  two;  hexaparental  chimeras  have  also  been 
produced. 


FUTURE 

Practical  applications  of  chimera  technolog\’  to 
livestock  are  not  obvious  at  this  stage  of  develop- 
ment. The  main  objective  of  this  research  is  to  pro- 
vide a genetic  tool  for  better  understanding  of  devel- 
opment, and  maternal-fetal  interactions. 

Recombinant  DNA 

DEFINITION 

The  introduction  of  foreign  DNA  into  the  germ- 
plasm. 

STATE  OF  THE  ART 

The  mechanics  of  changing  the  DN.A  molecules  of 
farm  animals  directly  have  not  yet  been  worked  out 
The  plasmid  methods  used  in  bacteria  may  not  he  a[>- 
plicable. 

FUTl'RE 

None  of  these  techniques,  no  matter  how  great  the 
potential,  will  he  of  any  use  in  animal  hreecling  until 
knowledge  of  genetics  is  greatly  adxanced  B<*fore 
one  can  alter  genes,  they  must  he  identifi(*d. 

Prior  to  exploitation  of  recombinant  DN.A  technol- 
ogy in  animal  breeding,  it  is  necessary  to  identify 
gene  loci  on  chromosomes,  i.e.,  genetic  m.ipping. 
Work  toward  this  goal  has  only  i(*c»*nlly  been  initi- 
ated and  rapid  progress  cannot  he  anticipated  Multi- 
variate genetic  determitiants  of  characteristics  ol 
economic  importance  are  antici|)at('d  to  he  tin*  rule 


Appendix  III-A 

History  of  the 
Recombinant  DNA  Debate 


The  hi.sloiA  i)t  tin*  over  the  ri.sks  from 

rl)\  A tet  hnu|iie>  aiui  the  (lov  ernment  > response 
m.i>  fk'  iIivuUhI  into  tour  phases  • Phase  I eoxered 
the  penotl  trom  ttie  lirst  awareness  ot  risks  to 
tuiinan  healtti  trom  t*\[)eriments  iinoKin^  reeomt)i- 
nant  I)\  A trDN.AI  in  thi*  summer  ot  1971  to  the  end 
ot  the  t onterenet*  at  the  \silomar  Center  in  Feh- 
ruar\  197">  w Itieti  r*‘sultetl  in  protot\  |)e  guidelines 
eo\ering  the  researeh  Phase  II  covered  the  period 
tmm  \silomar  through  the  il«‘\  elopment  h\  the  Na- 
tional Institutt's  ot  Health  (MHi  ot  the  Cuidelines  ot 
Jum*  l^t7ti  In  this  jxMiotl  the  puhlie  tirst  became 
sigmticantly  iiuoKed  in  the  ilehate  and  most,  it  not 
all  ot  the  |Hj|ic\  issues  were  clearK  Irametl  Phase 
III  tnim  mid  iy7l>  thnnigh  mid- 1978.  iinoKed  con- 
gressional consiileration  ot  the  issues  in  an  atmos- 
phere that  went  trom  almost  imminent  passage  ot 
legislation  to  the  cessation  ot  such  ettorts.  Phase  l\ 
covers  the  fxisllegislativ e period,  when  Mil  and  its 
organizational  parent  the  Department  of  Health. 
Kducation.  anil  Welfare  IHKW  ) (now  the  Department 
of  Health  and  Human  Services)  undertook  to  develop 
satisfactorv  voluntarv  standards  in  areas  over  w hich 
they  had  no  legal  authority  and  to  accommodate 
growing  pressure  for  public  involvement,  while 
av  oiding  a full  regulatory  role. 

Phase  I began  in  the  summer  of  1971.  w hen  sev- 
eral scientists  became  concerned  about  the  safety  of 
a proposeil  e.xperiment  to  insert  DN.A  from  S\40 
virus,  a monkey  tumor  virus  that  also  transforms  hu- 
man cells  into  tumor-like  cells,  into  a type  of  bacteria 
naturally  found  in  the  human  intestine.  After 
months  of  discussion,  the  scientist  who  had  pro- 
posed the  e.xperiment  decided  to  defer  it.  .Meanwhile, 
as  rD.N  A techniques  became  more  refined,  debates 
about  safety  increased:  at  the  June  1973  Gordon 
Research  Conference,  safety  issues  were  discussed. 
The  participants  voted:  to  send  a letter  to  the  Na- 
tional Academy  of  Sciences  (N.AS)  and  the  National  In- 


"For  a detailed  historv-  through  1977.  see  footnote  1.  For  a his- 
tory and  a discussion  of  the  broader  issues,  see  footnotes  2 and  3. 

'J  Swazev  J.  Sorenson,  and  C.  Wong.  Rislis  and  Benefits, 
Rights  and  Responsibilities:  A Historv'  of  the  Recombinant  D.\,A  Re- 
search Controversy  ' Southern  California  Law  Review  51:1019, 
September  1978. 

HT.  Grobstein.  .-1  Double  Image  of  the  Double  Helix  (San  Fran- 
cisco: VV  H.  Freeman  Co.  1979). 

^D.  Jaclison.  and  S.  Stich  (eds.l.  The  Recombinant  D\'A  Debate 
(Englewood  Cliffs,  .VJ.:  Prentice-Hall.  Inc.,  1979). 


stiUlte  of  .Medicine  requesting  the  appointment  of 
committees  to  study  potential  hazards  to  laboratory 
workers  and  the  public:  and  by  a narrow  majority* 
to  arrange  for  the  letter  to  be  published  in  the  widely 
read  journal.  Science,  to  alert  the  broader  scientific 
community.* 

•N.AS  appointed  a committee  of  prominent  scien- 
tists involved  in  rDN.A  research.  In  July  1974,  the 
[lanel  asked  for  a temporary  worldwide  moratorium 
on  certain  types  of  experiments,  and  called  for  an  in- 
ternational conference  on  potential  biohazards  of 
the  research  through  a letter  published  in  Science 
and  its  British  counterpart.  Nature.^  This  letter  also 
rei|uested  the  Director  of  NIH  to  consider  estab- 
lishing an  advisory  committee  to  develop  an  experi- 
mental program  to  evaluate  potential  hazards  and 
establish  guidelines  for  experimenters. 

In  response,  the  Director  of  NIH,  after  authoriza- 
tion by  the  Secretary  of  HEW,  established  the  Recom- 
binant DNA  .Molecule  Program  Advisory  Committee 
(later  renamed  the  Recombinant  DNA  Advisory  Com- 
mittee, RAC)  on  October  7,  1974,  along  the  lines  sug- 
gested by  the  NAS  Committee.  The  Committee’s 
charter  described  its  purpose  as:*’ 

The  goal  of  the  Committee  is  to  investigate  the  cur- 
rent state  of  knowledge  and  technology  regarding 
D.N.A  recombinants,  their  survival  in  nature,  and 
transferability  to  other  organisms;  to  recommend 
programs  of  research  to  assess  the  possibility  of 
spread  of  specific  DNA  recombinants  and  the  possible 
hazards  to  public  health  and  to  the  environment;  and 
to  recommend  guidelines  on  the  basis  of  the  research 
results.  This  Committee  is  a technical  committee,  estab- 
lished to  look  at  a specific  problem.  (Emphasis  added.) 
The  international  conference  called  for  by  the 
NAS  Committee  letter  was  held  at  the  Asilomar  Con- 
ference Center,  Pacific  Grove,  Calif.,  in  February 
1975.  The  organizing  committee  made  it  clear  that  its 
purpose  was  to  focus  on  scientific  issues  rather  than 
to  become  involved  in  considering  ethical  and  moral 
questions.  However,  in  one  session  the  few  lawyers 


“Swazev,  et  al.,  op.  cit.,  p.  1,023. 

^Letter  from  Maxine  Singer  and  Dieter  Soil  to  the  National 
Academy  of  Sciences  (NAS)  and  the  National  Institute  of  Medicine, 
reprinted  in  Science,  vol.  181,  1973,  p.  1114. 

^Letter  from  Paul  Berg,  et  al.  to  the  editor,  reprinted  in  Science, 
vol.  185,  1974,  p.  303. 

The  charter  of  the  Recombinant  DNA  Molecule  Program  Ad- 
visory Committee,  Oct.  7,  1974. 


315 


316  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


invited  confronted  the  scientists  with  some  of  these 
questions.®  The  conference  report  concluded  that  al- 
though a moratorium  should  continue  on  some  ex- 
periments, most  work  involving  rDNA  could  con- 
tinue with  appropriate  safeguards  in  the  form  of 
physical  and  biological  containment. 

In  Phase  II,  the  debate  widened  to  encompass 
broader  social  and  ethical  issues,  such  as  the  re- 
lationship between  scientific  freedom  of  inquiry  and 
the  protection  of  society’s  interests,  in  whatever 
manner  those  were  defined.  Such  issues  led  natural- 
ly to  questions  about  who  makes  the  decisions  and 
the  role  of  the  public  in  that  process.  Finally,  deci- 
sionmaking mechanisms  were  developed.  Issues 
raised  and  actions  taken  during  this  phase  in  many 
respects  controlled  the  subsequent  development  of 
the  Federal  response  to  the  debate,  and  created 
problems  that  continue  to  the  present.  At  this  stage, 
participation  in  the  debate  went  beyond  the  scien- 
tific community. 

Questions  of  ethics  and  public  policy  had  been 
raised  earlier,  but  they  now  received  much  wider  at- 
tention. On  April  22,  1975,  Sen.  Edward  M.  Kennedy, 
Chairman  of  the  Subcommittee  on  Health  of  the 
Senate  Committee  on  Labor  and  Public  Welfare,  held 
a half-day  hearing  on  science  policy  issues  arising 
from  rDNA  research.  In  May  1975,  a 2-day  con- 
ference on  "Ethical  and  Scientific  Issues  Posed  by 
Human  Uses  of  Molecular  Genetics"  was  held  under 
the  joint  sponsorship  of  the  New  York  Academy  of 
Sciences  and  the  Institute  of  Society,  Ethics,  and  the 
Life  Sciences.  In  addition  to  molecular  biologists,  par- 
ticipants included  lawyers,  sociologists,  psychiatrists, 
and  philosophers. 

The  issue  of  public  participation  arose  as  decision- 
making mechanisms  were  developed.  RAC  was  orig- 
inally composed  of  12  members  from  "the  fields  of 
molecular  biology,  virology,  genetics  and  microbiol- 
ogy.’®  Critics  first  noted  the  need  for  more  expertise 
in  the  fields  of  epidemiology  and  infectious  diseases, 
since  most  molecular  biologists  were  trained  as 
chemists.* *  RAC’s  membership  was  increased  to  16 
and  the  range  of  expertise  was  widened  to  include 
the  fields  of  epidemiology,  infectious  diseases,  and 
the  biology  of  enteric  organisms,  by  amendment  to 
the  charter  on  April  25,  1975. 

Since  some  members  were  conducting  the  re- 
search in  question,  critics  claimed  that  a conflict  of 
interests  existed.  They  also  noted  that  the  Committee 


“Swazey,  et  al.,  op.  cit.,  p,  1,034. 

^The  charter  of  the  Recombinant  DNA  Molecule  Program  Ad- 
visory Committee,  Oct.  7,  1974,  op.  cit. 

*One  of  the  members  of  the  original  RAC  (Stanley  Falkovv)  did 
have  substantial  expertise  with  enteric  organisms  and  £.  coli  in 
particular. 


advised  the  Director  of  NIH,  an  agency  whose  mis- 
sion was  to  foster  biomedical  research,  not  to  stop  or 
otherwise  regulate  it.  These  issues  were  brought  out 
in  a petition  to  NIH  signed  by  48  biologists  in  August 

1975.  Criticizing  a proposed  draft  of  the  guidelines  as 
setting  substantially  lower  safety  standards  than 
those  accepted  at  Asilomar,  the  petition  argued  for 
broader  representation  on  RAC  from  other  fields  of 
scientific  expertise  and  from  the  puhlic-at-large.  RAC 
itself  had  been  sensitive  to  these  limitations:  in  the 
summer  of  1975,  an  attempt  was  made  to  recruit 
nonscientists.'®  One  nonscientist  was  added  in 
January  1976,  and  another  was  added  in  .August 

1976. 

In  December  1975,  RAC  submitted  revised  dratt 
guidelines  to  the  Director  of  NIH,  Dr.  Donald 
Fredrickson.  Although  they  were  stricter  than  tho.se 
drafted  at  Asilomar,  some  criticized  them  as  being 
"tailored  to  fit  particular  experiments  that  are  al- 
ready on  the  drawing  hoards."”  I he  con.sensus  of 
RAC,  on  the  other  hand,  was  that  the  guidelines  were 
excessiv'ely  strict,  hut  that  it  was  in*cessary  to  lie 
overly  cautious  because  of  its  limited  exfiertise  in 
public  health,'^  In  any  event.  Dr.  Fri'derick.son  ar- 
ranged for  public  hearings  on  the  proposed  guide- 
lines at  a 2-day  meeting  in  February  1976  of  the  .Ad- 
visory Committee  to  the  Dii-ector,  a diviM  se  group  ol 
scientists,  physicians,  lawyers,  philsopliers,  and 
others.  A similarly  diver.se  group  of  scientists  and 
public  interest  advocates  wei'e  invited  to  attend 
Some  modifications  to  the  Guide-lines  [iroposed  hv 
Dr.  Fredrickson  as  a result  of  that  nu-eting  were 
adopted  and  others  were  rejecte-d  hv  It  At  in  .\pnl 
1976.'® 

The  final  major  issue  arising  during  this  period 
concerned  NIH's  lack  of  authority  to  set  condition.s 
on  research  funded  by  other  Federal  agencies  or  hv 
the  private  sector.  In  a June  2.  1976,  mi-eting  Im-- 
tween  Dr.  Fredrickson  and  .some  JO  re[)iesent.itiv  es 
of  industry,  including  pharmaci-utical  and  < hemical 
companies,  it  became  clear  that  some  rl).\  A rese.iri  h 
was  being  done;  however,  the  lepresent.itives  ap- 
peared hesitant  to  (aimmit  themselves  to  voluntai  v 
compliance  with  the  |)ro|)o.s(‘d  guidelines  '*  I he  jiri 


'“tJr.  hlizahclh  Kiitirr  ,i  liirmcr  H V(  iiii-inlM'i  imii 

munication.  Sept  II  I9H(1 

"N.  Wade,  "Recombinant  l),\  V Mil  s<•^^  sine  i Hull  s in  l.miu  h 
New  I echnologv',"  .S<(e/icf,  \ (il  19(1.197'.  pp  II".  1179 
‘'Kulter,  op.  cit 
'Mhid. 

''Subcommittee  on  Science  HcmmuIi  .ind  I r.  Iim .1. ,.l  lln- 
Hou.se  Committee  on  Scii‘nce  and  lei  lmuli.K\  i-rnrln  iti^^nrr’nn 
llumun  Cptwiics,  and  ( rll  Hiulnti\  l).\  t lln  nmhin.inl  ulr  Hr 

search  ISupp  Repoi't  III  94th  ( on^  3d  m-ss  ftri.  p l 


Appendix  III  A — History  of  the  Recombinant  DNA  Debate  *317 


mary  it'a-son  was  their  concern  over  protection  of 
pi-oprietarv  information 

Phase  II  culminated  with  the  pi'onuilgation  on 
June  23.  1976.  of  the  (Guidelines  for  Research  InvoK  - 
in^(  Recombinant  I).\  \ .Molecules  (1976  (Guidelines  ") 
covering  institutions  and  individuals  receiving  MM 
funils  for  this  research 

Phase  III  was  characterized  hv  attempts  to  remedy 
the  limited  applicability  of  the  (Guidelines.  Soon  after 
their  publication.  S«>nators  Kennedy  and  Ja\  its  sent  a 
letter  to  President  Ford,  calling  his  attention  to  the 
(Guidelines.  They  noted  that  any  risk  was  not  limited 
to  fetleralK  funded  research,  and  urged  him  to 
take  neces.sar\  steps  to  implement  the  (Guidelines 
throughout  the  research  community.  In  October 

1976.  the  S«*cretary  ol  lIF.W  with  the  appro\  al  of  the 
President  formed  the  Federal  Interagency  .Advisory 
Cornmittw  under  the  chairmanship  of  the  Director 
of  MM  to  determine  the  extent  to  which  the  (Guide- 
lines could  he  applied  to  all  research  and  to  rec- 
ommend necessary  executive  or  legislati\  e actions  to 
ensure  compliance  '•  In  .March  1977.  the  Committee 
concluded  that  existing  Federal  law  would  not  per- 
mit the  regulation  of  all  rO.VA  research  in  the  United 
States  to  the  extent  deemed  necessary: it  further 
recommendetl  new  legislation,  specifying  the  ele- 
ments of  that  legislation.'* 

During  1977  se\eral  bills  to  deal  with  this  and 
other  problems  were  introduced  in  Congress.  They 
addressed  in  different  ways  the  issues  of  the  extent 
of  regulatory  coverage,  the  mechanisms  for  regula- 
tion and  Federal  preemption  of  State  and  local  regu- 
lation The  major  bills  were  those  of  Rep.  Paul 
Rogers.  M R.  7897  (and  its  substitute,  H R.  11192)  and 
of  Sen.  Edw  ard  Kennedy,  S.  1217.* 

While  hearings  were  being  held,  three  devel- 
opments occurred  which,  by  the  end  of  1977,  had 
dissipated  much  of  the  impetus  for  legislation.  The 
first  was  the  expanded  role  of  R.AC.  On  September 
24.  1976.  its  charter  had  been  amended  once  more  to 
provide  for  additional  expertise  in  the  areas  of 
botany,  plant  pathologx',  and  tissue  culture.  More- 
over. its  membership  was  increased  from  16  to  20  so 
that  four  members  would  be  "from  other  disciplines 
or  representatives  of  the  general  public.”  This  was 
the  first  official  provision  for  public  representation 

’’Ibid.,  pp.  52. 

'^Interim  Report  of  the  Federal  Interagency  Committee  on  Recom- 
binant D\A  Research:  Suggested  Elements  for  Legislation,  Mar.  15, 

1977.  pp  3-4. 

'Mbid,.  pp.  9-10. 

'•Ibid.,  pp.  11-15. 

'For  a more  complete  discussion  of  the  legislation,  see  footnote 
19. 

‘•.  Recombinant  DN.A  .Molecule  Research,  " Congressional  Re- 
search Service,  issue  brief  ,\o.  IB  77024.  update  of  Jan.  2,  1979. 


although  two  nonscientists  vv^ere  already  members. 
The  number  of  nonscientists  remained  the  same 
until  December  1978.^°  Also,  RAC's  responsibilities 
were  defined  in  greater  detail,  including  the  respon- 
sibility tor  reviewing  large-scale  experiments.  Never- 
theless, RAC  continued  formally  at  least  to  be  "a  tech- 
nical committee,  established  to  look  at  a specific 
problem.” 

The  second  development  was  a growing  belief 
among  scientists  that  the  risks  of  the  research  were 
less  than  originally  feared.  This  was  based  on  the  fol- 
lowing: 1)  a letter  from  Roy  Curtiss  at  the  University 
of  Alabama  to  the  Director  of  NIH,  explaining  risk 
assessment  experiments  using  Escherichia  coli,  from 
which  he  concluded  that  the  use  of  E.  coli  K-12  host- 
vectors  posed  no  danger  to  humans;  2)  the  conclu- 
sions of  a committee  of  experts  in  infectious  diseases 
''  assembled  by  NIH  in  June  1977  in  Falmouth,  Mass., 
that  the  alleged  hazards  of  the  research  were  un- 
substantiated: and  3)  a prepublication  report  on  ex- 
periments showing  that  genetic  recombination  oc- 
curs naturally  between  lower  and  higher  life  forms, 
and  suggesting  that  the  rDNA  technique  was  not  as 
novel  as  presumed. 

The  third  dev'elopment  affecting  the  legislation 
was  a concerted  lobbying  effort  by  scientists  against 
what  they  considered  to  be  some  of  the  overly 
restrictive  provisions  of  the  bills,  especially  S. 
1217.^'  -phe  efforts  included  wide  circulation  of 

reports  (including  some  in  draft  form)  as  soon  as 
available,  which  supported  the  conclusion  that 
the  research  was  less  hazardous  than  originally 
supposed. 

By  the  end  of  1977,  the  legislation  was  in  limbo. 
This  situation  continued  in  early  1978,  although 
some  hearings  were  held.  On  June  1,  1978,  Senators 
Kennedy,  Javits,  Nelson,  Stevenson,  Williams,  and 
Schweiker  addressed  a letter  to  HEW  Secretary 
Joseph  Califano,  which  acknowledged  the  likelihood 
that  legislation  would  not  pass  and  urged  that  defi- 
ciencies in  the  regulatory  system  be  addressed 
through  executive  action  based  on  existing  authority, 
if  that  w'ere  to  be  the  case. 

During  Phase  IV,  NIH  and  its  parent  organization, 
HEW  (now  DHHS),  have  attempted  to  operate  in  the 
regulatory  vacuum  left  by  the  lack  of  legislation.  In 
response  to  the  consensus  that  developed  in  1977  on 


“William  Gartland,  Director  of  the  Office  of  Recombinant  DNA 
■Activities,  NIH,  personal  communication,  June  19,  1980. 

•‘B.  Culliton,  " Recombinant  DNA  Bills  Derailed:  Congress  Still 
Trying  to  Pass  Law,”  Science,  vol.  199,  Jan.  20,  1978.  pp.  274-277. 

^^D.  Dickson,  "Friends  of  DNA  Fight  Back,”  Nature,  vol.  272, 
April  1978,  pp.  664-665. 

“R.  Lewin,  "Recombinant  DNA  as  a Political  Pawn,”  New  Scien- 
tist, vol.  79,  Sept.  7,  1978,  pp.  672-674. 


318  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


the  question  of  risk,  RAC  proposed  revisions  to  the 
Guidelines,  which  placed  most  experiments  at  a 
lower  containment  level.  They  were  published  for 
public  comment  in  September  1977.* *  As  with  the 
original  Guidelines,  public  hearings  were  held  in  the 
course  of  a 2-day  meeting  of  the  Advisory  Committee 
to  the  Director  in  December  1977,  in  which  a diverse 
group  of  individuals  and  organizations  were  permit- 
ted to  comment.  However,  at  this  point,  HEW  took  a 
much  more  active  role  in  a situation  that  had  been 
handled  almost  entirely  by  NIH.^* 

When  RAC’s  charter  was  renewed  on  June  30, 
1978,  Secretary  Califano  reserved  the  power  to  ap- 
point its  members  instead  of  delegating  it  to  the 
Director  of  NIH  as  in  the  past.**  And  the  new  pro- 
posed Guidelines,  published  in  the  Federal  Register 
on  July  28,  1978,  were  accompanied  by  an  introduc- 
tory statement  by  Secretary  Califano  announcing  a 
60  day  public  comment  period  to  be  followed  by  a 
public  hearing  before  a departmental  panel  chaired 
by  HEW  General  Counsel  Peter  Libassi.***  The 
Secretary  was  particularly  interested  in  comments 
on:  new  mechanisms  to  provide  for  future  discre- 
tionary revision  of  the  Guidelines;  and  the  composi- 
tion of  the  various  advisory  bodies,  especially  the 
RAC  and  the  local  Institutional  Biosafety  Committees 
(IBCs).25 

The  public  hearing  called  for  by  Secretary  Cali- 
fano and  held  on  September  15,  1978,  was  a sig- 
nificant event  in  the  history  of  Federal  actions  on  the 
rDNA  issue.  Testimony  was  heard  from  represent- 
atives of  industry,  labor,  the  research  community, 
and  public  interest  groups;  more  than  170  letters  of 
comment  were  received  and  subsequently  reviewed. 
As  a result,  the  revised  final  Guidelines  of  December 
22,  1978,  were  significantly  rewritten  to  increase 
public  participation  in  the  decisionmaking  process:^® 

• Twenty  percent  of  the  members  of  the  IBCs  had 
to  represent  the  general  public  and  could  have 
no  connection  with  the  institution. 

• Most  of  the  records  of  the  IBCs  had  to  be  public- 
ly available. 

‘Shortly  thereafter,  in  October  1977,  the  Final  Environmental 
Impact  Statement  for  the  1976  Guidelines  was  published. 

^‘D.  Fredrickson,  "A  History  of  the  Recombinant  DNA  Guide- 
lines in  the  United  States,”  Recombinant  DNA  Technical  Bulletin, 
vol.  2,  July  1979.  pp.  87,  90. 

* ‘The  statement  providing  for  delegation  of  authority  that  ac- 
companied the  updated  Charter  was  not  signed  by  Califano.  See 
also,  footnote  24. 

“‘The  other  members  of  the  HEW  panel  were  Dr. 
Fredrickson,  Julius  Richmond,  who  was  the  Assistant  Secretary 
for  Health,  and  Henry  Aaron,  who  was  the  Assistant  Secretary  for 
Planning  and  Evaluation. 

“43  F.R.  33042,  July  28,  1978. 

“Statement  of  Secretary  Califano  accompanying  the  revised 
Guidelines,  43  F.R.  60080,  Dec.  22,  1978. 


• Major  actions,  such  as  decisions  to  except  other- 
wise prohibited  experiments  on  a case-by-case 
basis  or  to  change  the  Guidelines,  could  be  made 
only  on  the  advice  of  RAC  and  after  public  and 
Federal  agency  comment. 

The  increased  public  responsiveness  of  the  IBC’s 
was  crucial,  since  the  revised  Guidelines  placed  ma- 
jor responsibility  for  compliance  on  them.  This  had  i 
been  proposed  in  the  July  version  and  had  not  been  | 
changed  by  the  hearings.*  Califano  also  announced 
he  would  appoint  14  new  members  to  the  RAC,  in- 
cluding people  knowledgeable  in  fields  such  as  law, 
public  policy,  ethics,  the  environment,  and  public 
health. ^^^**  All  of  these  changes  were  envisioned  to 
"prov'ide  the  opportunity  for  those  concerned  to 
raise  any  ethical  issues  posed  by  recombinant  DNA  f 
research”  and  to  change  the  role  of  the  R.AC  to  "ser\  e 
as  the  principal  advisory  body  to  the  Director  of  Mil 
and  the  Secretary  of  HEW  on  recombinant  DNA 
policy.”^®***  ) 

In  addition  to  broadening  public  participation,  | 
Califano  attempted  to  deal  with  a major  limitation  of  | 
the  Federal  response— the  (iuidelines  did  not  co\er  ‘ 
private  research.  He  directed  the  Food  and  Drug  .Ad- 
ministration (FDA)  to  take  steps  to  require  that  any 
firm  seeking  approval  of  a product  rec|uiring  the  use 
of  rDNA  techniques  in  its  de\elopment  or  manu- 
facture, demonstrate  compliance  with  the  Guidelines 
for  the  work  done  on  that  product;  an  FDA  notice  of 
its  intention  to  propose  such  regulations  accom- 
panied the  revised  Guidelines  in  the  Federal  K»‘gister.  .1 
In  addition,  he  requested  the  Kn\  ironmental  I’rotec- 
tion  Agency  (EPA)  to  review  its  regulatory  authority 
in  that  area.  He  believed  if  both  agencies  could 
regulate  research  on  products  within  their  juiisdic 
tion,  "virtually  all  recombinant  DN.A  re.search  in  this 
country  would  he  brought  under  the  re(|uirements 
of  the  revised  guidelines.  In  the  nuMUtime.  the 


‘As  pari  of  th(>  r(‘visioti  process,  Hf.W  held  .1  meeting  m (K  lobin 
1978  for  IBG  chairper.sons  in  order  lo  exchange  miorm.ilion  and 
experiences  gaineil  under  the  1976  (,uidelines 

“Ibid. 

* ‘ This  was  implemenled  by  an  amendmiml  lo  ihe  R \(  ( barli'r 
on  Dec.  28.  1978,  which  increased  ibe  membersinp  lo  2.'  and 
changed  Ibe  composition  lo  the  lolliivxing  calegorn-s  II  al  Ir.isi 
eight  specialists  in  moleculai-  biologx'  or  rl).\  \ rescan  b 21  al  least 
six  specialists  in  other  scientific  lields  ,ind  :ll  al  le.isl  six  persons 
knowledgeable  in  laxv,  |uiblic  policy,  Ibe  enx  ironmeni  and  piiblii 
or  occupational  health  In  addition,  the  ( harlei  xx.is  amended  to 
grant  nonvoting  representation  lo  1 epi  esenlalix  es  ol  xanoos  li-d 
eral  agencies. 

“Ibid. 

“‘Thi’  Charter  was  nexer  amended  lo  1 h.ingi’  01  deli-li-  iIm- 
final  sentence  of  the  'I’urpose  section  xxbii  h stales  Ibis  ( om 
mittee  is  a technical  commillee  established  lo  look  al  a s|m-»  ilu 
problem." 

“Ibid.,  p 6111181 


Appendix  lll-A— History  of  the  Recombinant  DNA  Debate  • 319 


revised  Guidelines  provided,  for  the  first  time,  for 
voluntary  registration  of  projects  with  NIH,  in  which 
the  registrant  would  agree  to  abide  only  by  the  con- 
tainment standards  of  the  Guidelines.*’ 

Other  major  changes  were  embodied  in  the  new 
Guidelines.  Because  of  the  consensus  that  the  ex- 
periments posed  lower  risks  than  originally  thought, 
some  types  of  experiments  were  exempted,  while 
containment  levels  were  lowered  for  almost  all 
others.  In  order  to  provide  greater  tlexibility,  these 
Guidelines  permitted  exceptions  on  a case-by-case 
basis,  and  included  procedures  for  their  change  on  a 
piecemeal  basis  without  going  through  the  whole  in- 
ternal process  at  HEW  . For  major  changes,  the  pro- 
cedure was;  1)  publication  of  the  proposed  changes 
in  the  Federal  Register  at  least  30  days  prior  to  a R,\C 
meeting;  2)  R.AC  consideration  of  the  proposed 
changes:  and  3)  publication  in  the  Federal  Register  of 
the  final  decision  of  the  Director,  N'lH.  The  standard 
for  all  actions  of  the  Director  under  the  Guidelines 
was  "no  significant  risk  to  health  or  to  the  environ- 
ment.’’** Lastly,  the  new  Guidelines  delegated  project 
approx  al  to  the  IBCs. 

The  problems  posed  by  voluntary  compliance  and 
commercialization  haxe  continued  to  be  addressed 
by  MH.  In  a second  major  revision  to  the  Guidelines 
on  January  29.  1980,  a section  (Part  \ I ) was  added  to 
specify  procedures  for  voluntary  compliance.  * * On 


Iconiinuedfrom  p.  3tH) 

•Subsequently.  Califano  sent  similar  letters  to  the  Secretaries  of 
.Agriculture  (February  1979)  and  Labor  (July  1979)  requesting 
them  to  consider  how  their  agencies'  authorities  could  be  used  to 
require  prixate  sector  rDN.A  research  to  comply  with  the 
Guidelines.” 

"Minutes  of  the  Interagency  Committee  on  Recombinant  DNA 
Research,  p.  3,  July  17.  1979,  reprinted  in  Recombinant  DNA  Re- 
search, vol.  5.  p 132.  et.  seq. 

’’See.  l\'-F-3.  1978  Guidelines. 

“Sec.  IX  -E-l-b. 

■ ■ Sev  eral  responses  to  the  FD.A  notice  had  questioned  the  agen- 
cy s legal  authority  to  regulate  prixate  rDN.A  research.  Conse- 
quently, Dr.  Fredrickson  and  Dr.  Donald  Kennedy,  then  Commis- 
sioner of  Food  and  Drugs,  developed  a draft  supplement  to  the 
Guidelines,  specifying  procedures  for  voluntary  compliance  by  in- 
dustry. It  was  published  for  comment  on  .Aug.  3,  1979  (44  F.R. 
45868)  and  incorporated  as  part  of  the  proposed  revised  Guide- 
lines of  November  30,  1979.  (44  F.R.  69210,  69247). 


April  11,  1980,  NIH  published  Physical  Containment 
Recommendations  for  Large  Scale  Uses  of  Organisms 
Containing  Recombinant  DNA  Molecules  in  the  form 
of  Draft  Part  \'II  to  the  Guidelines.**  Besides  setting 
large  scale  containment  levels,  this  document  recom- 
mends that  the  institution:  appoint  a biological  safety 
officer  with  specified  duties;  and  establish  a worker 
health  surveillance  program  for  work  requiring  a 
high  (Pj)  containment  level.  Finally,  a more  ad  hoc  re- 
quirement has  been  used  since  October  1979  for  ap- 
prox'als  of  industrial  requests  for  cultures  up  to  750 
liters  (1);  the  approvals  were  conditioned  on  NIH 
designated  observers  being  permitted  by  the  com- 
panies to  inspect  their  facilities.*'*  At  least  one  inspec- 
tion has  taken  place. 

On  November  21,  1980,  NIH  adopted  the  third  ma- 
jor revision  to  the  Guidelines.**  It  contained  these 
significant  changes:  institutions  sponsoring  the 
research  are  no  longer  required  to  register  their 
projects  xvith  NIH  pursuant  to  an  informational  docu- 
ment called  a Memorandum  of  Understanding  (MUA) 
xvhenever  the  containment  levels  are  specified  in  the 
Guidelines;  and  NIH  will  no  longer  review  IBC  deci- 
sions on  experiments  for  which  containment  levels 
are  specifieid  in  the  Guidelines. 

On  November  21,  1980,  NIH  also  promulgated 
revised  application  procedures  for  large-scale  pro- 
posals. The  application  must  include  the  following  in- 
formation: 1)  the  registration  document  submitted  to 
the  local  IBC;  2)  the  reason  for  wanting  to  exceed  the 
10-1  limit;  3)  evidence  that  the  rDNA  to  be  used  was 
rigorously  characterized  and  free  of  harmful  se- 
quences; and  4)  specification  of  the  large-scale  con- 
tainment level  proposed  to  be  used  as  defined  in  the 
NIH  Physical  Containment  Recommendations  of 
April  n,  1980. 


In  addition  to  adding  part  VI  to  the  Guidelines,  the  most  signifi- 
cant change  in  the  January  1980  Guidelines  was  the  addition  of 
sec.  III-O,  which  permitted  most  experiments  using  E.  coli  K-12 
host-vector  systems  to  be  done  at  the  lowest  containment  levels. 

”45  F.R.  24968,  Apr.  11,  1980. 

”44  F.R.  69251,  Nov.  30,  1979. 

”45  F.R.  77372,  Nov.  21,  1980. 


Appendix  III-B 

Constitutional  Constraints 

on  Regulation 


Under  the  checks  and  balances  of  our  system  of 
government,  the  Constitution,  as  ultimately  inter- 
preted by  the  Supreme  Court,  requires  certain  pro- 
cedural and  substantive  standards  to  be  met  by  stat- 
utory or  other  regulation  imposed  upon  an  activity. 
These  requirements  depend  on  the  nature  of  the  ac- 
tivity involved.  In  the  present  case,  it  will  be  useful  to 
consider  first  the  regulation  of  basic  research  and 
then  the  regulation  of  technological  applications, 
such  as  the  production  of  pharmaceuticals  by  using 
genetic  engineering  methods. 

Research 

With  respect  to  research,  the  fundamental  ques- 
tion is  what  limitations,  if  any,  may  be  placed  on  the 
search  for  scientific  knowledge.  The  primary  appli- 
cable constitutional  provision  is  the  first  amendment, 
which  has  been  broadly  interpreted  by  the  Supreme 
Court  to  severely  limit  intrusion  by  the  Government 
on  all  forms  of  expression.*  ^ ® Another  constitutional 
safeguard,  known  as  equal  protection,  is  secondarily 
involved. 

If  the  Supreme  Court  were  to  recognize  a right  of 
scientific  inquiry,  its  boundaries  would  not  exceed 
those  for  freedom  of  expression. “ There  is  disagree- 
ment among  commentators  on  this  issue  concerning 
the  boundaries  of  the  first  amendment,^  and  certain- 
ly disagreement  on  the  application  of  generally  ac- 
cepted principles  to  particular  cases.  Moreover, 
there  have  been  no  judicial  decisions  dealing  with  the 
precise  issue  at  hand.  However,  it  is  possible  to  out- 
line general  principles  derived  from  judicial  deci- 
sions interpreting  the  first  amendment,  and  indicate 
how  they  might  be  applied  by  the  courts  to  attempts 
to  regulate  genetic  research. 

There  are  very  few  limitations  on  the  written  or 
spoken  word.  The  prohibitions  against  obscenity  or 
"fighting  words”*  clearly  would  be  inapplicable  here. 

'Harold  P.  Green,  "The  Boundaries  of  Scientific  Freedom"  Regulation  of 
Scientific  Inquiry:  Societal  Concerns  With  Research,  Keith  M.  VVuIff  (ed.) 
(Washington,  D.C.:  AAAS,  1979),  pp.  139-143. 

^Thomas  1.  Emerson,  "The  Constitution  and  Regulation  of  Research,"  Reg- 
ulation of  Scientific  Inquiry:  Societal  Concerns  With  Research,  Keith  M.  VVuIff 
(ed.)  (Washington.  D.C.:  AAAS,  1979),  pp.  129-137. 

'John  A.  Robertson,  "The  Scientists'  Right  to  Research:  A Constitutional 
Analysis,"  Southern  California  Law  Review  51.1203,  September  1978. 

"Green,  op.  cit.,  p.  140. 

'Emerson,  op.  cit.,  pp.  131-134. 

•"Fighting  words"  are  those  provoking  violent  reaction  or  imminent 
disorder. 


For  many  years,  the  Supreme  Court  has  conceptual- 
ized the  right  of  free  expression  in  terms  of  a market- 
place of  ideas— through  the  open  and  full  discussion 
of  all  ideas  and  related  information,  the  valuable, 
valid,  or  useful  ones  will  be  accepted  by  society, 
while  the  ridiculous  or  even  dangerous  ones  will  be 
so  demonstrated  and  discarded.  This  is  a consensual 
process;  no  person,  group,  or  institution  has  suffi- 
cient wisdom  to  prejudge  ideas  and  deny  them 
admittance  to  that  intellectual  marketplace,  even  if 
they  threaten  fundamental  cultural  values,  for  such 
values,  if  worthwhile,  will  survive.  Under  this  con- 
cept, scientists  would  certainly  have  virtually  unre- 
strained freedom  to  think,  speak,  and  write. 

Difficulties  arise  with  actions,  such  as  experimen- 
tation, which  may  be  essential  to  the  implementation 
of  freedom  of  expression.  Recent  Supreme  Uourt 
cases  have  recognized  a limited  protected  interest  of 
the  media  to  gather  information  as  an  essential  ad- 
junct to  freedom  of  publication.  By  analogv',  it  may 
be  argued  that  scientists  would  also  be  protected  in 
their  research,  as  a necessary  adjunct  to  freedom  of 
expression.  On  the  other  hand,  the  information 
gathering  cases  usually  involve  access  to  Govern- 
ment facilities,  such  as  courtrooms  or  prisons.  I hey 
are  based  on  the  principle  that  actions  by  the  (lov- 
ernment  should  he  open  to  [jublic  scrutiny— a con- 
cept not  directly  ap[)licable  to  the  |)ce.sent  issue 
More  importantly,  the  Cx)urt  has  long  recognizeil 
that  actions  related  to  expre.ssion  can  be  regulated 
and  that  regulation  may  increase  u ith  the  degree  ol 
the  action's  impact  on  people  or  the  environment 
The  Court  would  probably  ap|)ly  what  has  been 
called  a structured  balancing  test:'*  i.e,,  regul.ilion 
would  be  deemed  valid  only  when  the  (lovei  iiment 
sustains  the  burden  of  |)rov  ing:  1)  that  there  are 
'compelling  reasons”  for  the  regulation:  and  2)  that 
the  objective  cannot  be  achieved  by  less  diasiic 
means,”  i.e.,  by  more  narrowly  dratted  regulations 
having  less  ini|)act  on  first  amendment  rights 

The  secvjnd  part  of  the  test  is  fairly  straightlor 
ward.  Govei  nmental  restrictions  must  he  kept  to  ,i 
minimum.  F.g.,  where  possible,  they  should  be  leg 
ulatory  rather  than  prohibitory,  temporarv  r.ither 
than  permanent,  involve  the  least  burden,  .ind  soon 
rbe  difficult  |)art  of  this  test  lies  in  determining 


"Ibid.,  |)  134 


320 


Apppendix  lll-B— Constitutional  Constraints  on  Regulation  • 321 


u hat  is  a i-om[)elling  reason.  I he  [)rotection  of  health 
oi-  the  em  ironment  is  the  most  clearlv  acceptable 
reason  tor  regulation.  In  aclditiott.  the  protection  of 
incli\  idual  rights  and  [tersonal  dignity  is  generally 
consiilered  an  acceptable  reason,  f^.g..  the  National 
Research  Act*  re(|uires  that  all  biomedical  and  be- 
havioral researih  iiuohing  human  subjects  sup- 
[)orteil  under  the  Public  Health  Service  .Act  he  re- 
viewed In  an  Institutional  Review  Board  in  order  to 
[)rotect  the  rights  and  w elfare  of  the  subjects. 

rhe  alKJve  discussion  relates  to  protection  from 
physical  risks  due  to  the  process  of  research.  Could 
the  (k)vernment  regulate  or  forbid  e.xperimentation 
solely  because  the  product  (knowledge)  threatens 
cultural  V alues  or  other  intangibles  such  as  the  genet- 
ic inheritance  of  mankind?  Religious  or  philosophical 
objections  to  research,  based  solely  on  the  rationale 
that  there  are  some  things  mankind  should  not 
know,  contlict  with  the  basic  principles  of  freedom 
of  e.xpression  and  would  not  he  sufficient  reason  on 
constitutional  grounds  to  justify  regulation.  Even  if 
the  rationale  underlying  this  objection  were  e.xpand- 
ed  to  include  situations  w here  know  ledge  threatens 
fundamental  cultural  values  about  the  nature  of 
man.  control  of  research  for  such  a reason  probably 
would  not  be  constitutionally  permissible.  The  ra- 
tionale w ould  again  conflict  w ith  the  marketplace  of 
ideas  concept  that  is  central  to  freedom  of  e.xpres- 
sion. However,  w hat  if  the  knowledge  were  to  pro- 
vide the  means  to  alter  the  human  species  in  such  a 
way  that  the  physical,  psychological,  and  emotional 
essence  of  what  it  is  to  be  human  could  be  changed? 
No  precedent  exists  to  prov  ide  guidance  in  determin- 
ing an  answer.  Were  the  situation  to  arise,  the 
Supreme  Court  might  fashion  another  limitation  on 
the  concept  of  free  e.xpression  in  the  same  way  it 
developed  the  obscenity  or  "fighting  words"  doc- 
trines. 

The  discussion  thus  far  has  had  as  its  premise  a 
direct  regulator}  approach  to  research.  There  is  a 
more  indirect  approach,  which  would  be  constitu- 
tionally permissible  and  could  accomplish  much  of 
w hat  direct  regulation  might  attempt,  including  pre- 
vention of  the  acquisition  of  some  forms  of  knowl- 
edge. This  is  the  use  of  the  funding  power.  The 
lifeblood  of  modern  science  in  the  United  States  is 
the  Federal  grant  system.  Yet  it  is  generally  agreed 
that  Government  has  no  constitutional  duty  to  fund 
scientific  research.®  This  is  a benefit  voluntarilv  pro- 
vided to  which  many  kinds  of  conditions  may  be  at- 
tached. The  only  consitutional  limitation  on  such  an 
approach  would  be  the  concept  of  equal  protection— 
any  restrictions  must  apply  to  aU  or  must  not  be  ap- 


■ Public  Law  93-J48  (1974),  42  U.S.C.  §289  1-3. 
•Green,  op.  dt.  p.  141. 


plied  in  a discriminatory  way  without  compelling 
reasons. 

Congress  could  therefore,  mandate  by  law  that 
certain  kinds  of  research  not  be  funded  or  be  con- 
ducted in  certain  ways.  .An  example  is  the  National 
Research  Act,  discussed  previously.  However,  this 
approach  may  have  some  serious  practical  limita- 
tions because  of  the  difficulty  of  determining  which 
molecular  biological  research  might  lead  to  the  pro- 
scribed knowledge.  Much  discretion  would  have  to 
be  left  to  the  funding  agency,  which  is  likely  to  be  un- 
sympathetic or  even  hostile  to  such  an  approach,  if  it 
V iews  its  primary  mission  as  fostering  research. 

Applications  and  products 

.Although  fears  have  been  expressed  that  current 
genetic  technologies  may  lead  to  applications  that 
would  be  detrimental,  no  one  can  reasonably  con- 
clude, at  the  present  time,  that  this  will  actually  oc- 
cur. For  this  reason,  the  most  constitutionally  per- 
missible approach  in  all  probability  will  be  to  regu- 
late the  applications  of  the  science.  In  such  situa- 
tions, whatever  harms  occur  tend  to  be  more  tangi- 
ble and  the  governmental  interests,  therefore,  more 
clearly  defined.  Moreover,  since  fundamental  con- 
stitutional rights  are  generally  not  involved,  statutes 
and  regulations  are  subjected  to  a lower  level  of 
scrutiny  by  the  Federal  courts. 

The  constitutional  authority  for  Federal  regulation 
of  the  applications  of  technologies  such  as  genetic  en- 
gineering lies  in  the  commerce  clause,  article  I,  sec- 
tion 8 of  the  Constitution,  which  grants  Congress  the 
power  'To  Regulate  Commerce  wath  foreign  Nations, 
and  among  the  Several  States.”  In  contrast  to  sit- 
uations involving  fundamental  rights,  the  Supreme 
Court  has  interpreted  this  clause  as  ghing  Congress 
extremely  broad  authority  to  regulate  any  activity  in 
any  way  connected  with  commerce.  It  has  been  vir- 
tually impossible  for  Congress  not  to  find  some  con- 
nection acceptable  to  the  courts  between  commerce 
and  the  goals  of  a particular  piece  of  legislation.  * The 
standard  of  review  of  such  legislation  by  the  Federal 
courts  is  to  determine  if  it  bears  a rational  re- 
lationship to  a valid  legislative  purpose.  If  so,  the 
Court  vv'ill  uphold  the  legislation  and  will  not  second 
guess  the  legislators.  This  standard  of  review  rec- 
ognizes that  a statute  results  from  the  balancing  of 
competing  interests  and  policies  by  the  branch  of 
Gov'ernment  created  to  function  in  that  manner. 

*See  Wickard  v.  Filbum,  3l7  U.S.  Ill  0942)  in  which  the  Supreme  Court  up- 
held civil  penalties  for  violation  of  acreage  allotments  established  by  the 
.Agricultural  .Adjustment  .Act  of  1938,  covering  the  amount  of  wheat  that  in- 
dividual farmers  could  plant,  even  if  the  wheat  was  intended  for  self-con- 
sumption. The  rationale  was  that  even  though  the  indit  idual  farmer  s wheat 
had  no  measurable  impact  on  interstate  commerce.  Congress  could  prop- 
erly determine  that  all  wheat  of  this  category,  if  exempted  from  regulation, 
could  undercut  the  purpose  of  the  .Act,  which  was  to  increase  the  price 
farmers  received  for  their  various  crops. 


\ 

\ 

Appendix  III-C 

Information  on  International  i 

% 

Guidelines  for  Recombinant  DNA  i 


The  following  information  is  based  largely  on  in- 
ternational surveys  undertaken  by  The  Committee 
on  Genetic  Experimentation  of  the  International 
Council  of  Scientific  Unions  reported  as  of  July 
1979.1 

/.  Nations  that  had  established  guidelines  for  conduct 
of  rDNA  research  or  were  using  the  guidelines  of 
other  nations: 


Australia 

Italy 

Belgium 

Japan 

Brazil 

Mexico 

Bulgaria 

Netherlands 

Canada 

New  Zealand 

Czechoslovakia 

Norway 

Denmark 

Poland 

German  Democratic 

South  Africa 

Republic 

Sweden 

Federal  Republic  of 

Switzerland 

Germany 

Taiwan 

Finland 

United  Kingdom 

France 

United  States 

Hungary 

U.S.S.R. 

Israel 

Yugoslavia 

II.  Nations  that  had 

not  established  guidelines  o. 

not  responded  with  updated  information: 

Country 

Yes  No 

Austria 

X 

Ghana 

X 

India 

X 

Iran 

X 

Jamaica 

X 

Korea 

X 

Nigeria 

X 

Singapore 

X 

Sri  Lanka 

X 

Sudan 

X 

Turkey 

X 

III.  Nations  that  had  drafted  their  own  guidelines: 

Canada 

Japan 

Federal  Republic  of 

United  Kingdom 

'Report  to  COGENE  from  the  working  group  on  Recombinant  DNA  Guide- 
lines, May  1980. 


Germany  United  States 

France  U.S.S.R. 

Italy 

/V.  Nations  that  had  modified  the  guidelines  of  other, 
indicated,  countries: 


Australia  (UK,  U.S.) 
Belgium  (UK,  U.S.) 

Brazil  (U.S.) 

Bulgaria  (U.S.S.R.,  U.S.) 
Czechoslovakia  (U.S.S.R., 
U.S.,  Fed.  Rep.  Ger.) 
Denmark  (UK) 

East  (ierman  Democratic 
Republic  (UK,  U.S., 
Netherlands) 

Finland  (U.S.  mainly) 
Hungary  (U.S.) 


Mexico  (U.S.) 
Netherlands  (U.S.) 
New  Zealand 
Norway  (U.S.) 
Poland  (U.S.) 

South  Africa  (U.S.) 
Sweden  (U.S) 
Switzerland  (U.S.) 
Taiwan  (IfS.,  UK) 
Yugoslavia 
(European  Science 
Foundation) 


V.  Nations  in  which  entirely  voluntary  guidelines  have 
been  adopted: 

Finland 


VI.  Nations  with  guidelines  that  are  enforceable 
through  control  of  research  funding: 


Australia^ 

Canada 

Czechoslovakia^ 

Denmark 

Federal  Republic  of 
Germany^ 
France 

German  Democractic 
Republic 


Japan 

Netherlands'* * 

Norway 

South  Africa 

Sweden 

Switzerland 

'Faiwaiv' 

United  Kingdom' 
United  States 


i 


! 

1 


^■'Submissions  may  be  made  directly  to  the  Ai  adeni)  of  Si  leni-e  or 
through  a granting  agency.  In  the  latter  casiv  it  is  .i  iTi|uiremeni  (or  the  «p 
plicant  to  observe  the  recommendations  o(  the  Ar.idrms  s standing  ( oni 
mittee  if  the  agency  makes  a grant  for  the  work  Otherwiv  lire  giiidrlinn 
are  voluntary  with  the  worker  reipiiriil  to  make  an  annual  rejiorl  on  prog 
ress,  or  more  frequently  if  conditions  o(  the  esiii'rimi-nl  Ism  h at  wilunml  4 
are  changed  appreciably  " 

*l"Control  through  Academy  of  Sciences  and  Minisirs  ol  Health 
i-Several  research  organizations  require  rii ewers  o(  grants  to  appK  lt*r 
NIH  guidelines  until  their  own  national  guidelines  are  eonipletiil 
i*The  Netherlands  Organization  for  the  Adsani-einenI  of  ISire  Ri-teair  h 
will  only  subsidize  projects  which  have  bii-n  given  the  i ommiltee  1 1 ontrot 
^"Waiting  for  response  from  National  Adv  isory  ( ommillrf 
^''Notification  of  proposals  to  (,M  \(,  bm  ami- compultvirv  kiigiiti  I 1978 
In  addition,  funding  iHidies  re<|uire.  as  a condition  of  liinifiog  tiMMis  ad 
vice  to  be  sought  and  followed 


322 


1 


Appendix  lll-C  Information  on  International  Guidelines  for  Recombinent  DNA  • 323 


VII.  \ations  in  which  guidelines  are  legally  en- 
forceable: 

Hungary 

L'.S.S.R.' 

Finland  '.At  present,  the  guidelines  are 

entirely  voluntary,  but  in  the  near 
tuture,  the  intention  is  to  include 
them  in  the  law  of  infectious  dis- 
eases w hen  they  u ill  become  legally 
enforceable." 

South  Atrica , At  present  the  guidelines  are  not 
legally  enforceable.  They  will  only 
become  so  if  regulations  under  the 
existing  Health  Act  of  1977  and  the 
Animal  Diseases  and  F'arasites  .Act  of 
1956  are  promulgated:  and  none  are 
intended  at  present." 

United  Kingdom  The  regulation  to  notify  GM.AG 
does  not  strictly  mean  that  the 
Williams  Guidelines  themselves  are 
legally  enforceable.  But,  under  the 
Health  and  Safety  at  Work  ,Act 
(w  ithin  w hich  the  Regulations  were 
introduced),  it  is  e.xpected  that  ac- 
count w ill  be  taken  of  the  relevant 
Codes  of  Practice  and  the  advice 
given  by  GM.AG.” 

Mil.  \ations  in  w hich  observance  of  the  guidelines  is 
monitored  by  a nationally-directed  mechanism: 


.Australia 
Czechoslo\  akia 
German  Democratic 
Republic 
France 
Hungary 
Japan 


Norway 
South  .Africa 
Sweden 

United  Kingdom 
United  States 
U.S.S.R. 
Vugosla\ia 


IX.  Xations  in  which  a license  or  other  authorization 
for  recombinant  DXA  activity  is  granted: 


—to  an  institution:  U.S.S.R. 

—to  an  indixdual  laboratory:  Hungary,  Czechoslo- 
vakia 

—to  an  indix'idual  scientist:  .Australia,  Canada,  Ger- 
man Democratic  Republic,  Federal  Republic  of 
Germany,  Finland,  France,  Japan,  Norway,  South 
.Africa,  Sweden,  United  Kingdom®,  United  States 
and  U.S.S.R. 

Netherlands:  ‘There  are  gentlemen’s  agreements, 
signed  by  the  indix  idual  scientist,  the  institution 
and  the  Committee.”  The  reports  of  the  Committee 
also  recommend  legislation  that  w'ill  require  regis- 
tration of  research  projects  in  this  field  and  make 
binding  the  guidelines  and  supervision  of  their 
observance.  (Report  of  the  Committee  in  Charge  of 


the  Control  on  Genetic  Manipulation,  Amsterdam, 
March  1977,  p.  54.) 

Bulgaria,  Switzerland:  None  of  the  above. 

Taiwan:  No  response. 

“The  Group  advises  on  proposals  from  individual  workers,  but  considers 

them  in  the  conte.xt  of  information  about  tbe  centre’  in  which  the  work  is  to 

go  on.” 

X.  Xations  in  which  special  provisions  for  agriculture 
and/or  industrial  research  and  applications  have 
been  made: 

Czechoslovakia.  "10  liter  maximum  volume  of  the 
culture  containing  recombinant 
DNA” 

German 

Democratic 

Republic "The  GDR  Guidelines  will  be  com- 

pulsory for  industrial  and  agricul- 
tural applications.  10-liter  maximum 
deviations  may  be  allowed  by  the 
Minister  of  Health  if  suggested  by 
the  Committee.” 

Federal  Republic 

of  Germany  . . "Specification  of  containment  of 


plants” 

France “Industry,  maximum  volume  of  cell 

culture  is  set  at  10  liters” 

Norway "The  Guidelines  cover  both  agri- 


culture and  industry.  Application  of 
recombinant  DNA  research  outside 
an  approved  laboratory  is  prohib- 
ited. Otherwise  the  Committee  fol- 
lows the  NIH  Guidelines.” 

United  Kingdom  "Agriculture,  industry;  see  Williams 
Report,  paragraphs  1.3,  2.7,  5.13 
and  appendix  II,  section  34.” 

United  States  . . "Agriculture.  NIH  Guidelines  pro- 
vide containment  levels  for  cloning 
total  plant  DNA,  plant  virus  DNA 
and  plant  organelle  DNA  in  E.  coli 
K-12,  and  provide  general  guidance 
for  the  use  of  plant  host-vector  sys- 
tems. 10  liter  maximum.  A proposed 
Supplement  to  the  Guidelines  for 
voluntary  compliance  by  the  private 
sector  is  under  consideration  by 
RAC.  Development  of  a monograph 
for  large-scale  applications  has  been 
proposed.” 

U.S.S.R "Guidelines  are  compulsory  for  in- 

dustrial and  agricultural  applica- 
tions. 10  liter  maximum.  Deviation 
is  allowed  by  the  Recombinant  DNA 
Commission.” 

Other 

respondents . . No 


324  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


XL  Number  of  laboratories  currently  engaged  in  re- 
combinant DNA  activities: 


Country 

Any  labs? 

How  many? 

Australia 

yes 

16 

Austria 

no^" 

Belgium 

yes- 

6 

Brazil 

yes 

5 

Bulgaria 

yes- 

no  response 

Canada  

yes 

10-15 

Czechoslovakia 

yes 

3 

Denmark 

German  Democratic 

yes- 

several 

Republic 

Federal  Republic 

yes 

5 

of  Germany 

yes 

10-20 

Finland 

yes 

3 (3-4  planned) 

France 

yes- 

12 

Ghana  

no- 

Hungary 

yes- 

1-2 

India 

no- 

Iran 

no- 

Israel 

yes- 

1 

Jamaica 

no- 

Japan  

yes 

35 

Korea  

no- 

Netherlands 

yes 

7 

New  Zealand 

yes 

2 

Nigeria 

no 

Norway 

yes 

not  stated 

Philippines 

no- 

Poland 

yes- 

3 

Singapore 

no- 

South  Africa 

yes- 

3 

Sri  Lanka 

no- 

Sudan  

no- 

Sweden 

yes- 

2 

Switzerland 

yes- 

18 

Taiwan 

yes 

2 

Turkey  

no- 

United  Kingdom 

yes 

45 

United  States 

yes- 

50 

U.S.S.R 

yes- 

6 

Yugoslavia 

yesb 

4 

XII.  Countries  in  which  specific  training  for  workers 
and  safety  officers  in  recombinant  DNA  activities  is 
required  by  the  guidelines: 


Country  Yes  No  Other 

Australia ^ 

Bulgaria X 

Canada X 

Czechoslovakia.  X^ 

German 

Democratic 

Republic X'= 

Federal  Republic 
of  Germany  . . X^ 

Finland X 

France X 

Hungary X 

Japan X 

Netherlands  ...  X« 

Norway Xf 

South  Africa ...  X 

Sweden X 

Switzerland  . . . "recommended  ” 

Taiwan "recommended” 

United  Kingdom  Xs 

United  States  . . X'> 

U.S.S.R X 

Yugoslavia  ....  no  response 


Other  respondents:  no  or  no  response  to  question. 


^Australia:  "Require  expertise  through  Biosafety  ( ommittee 

^Czechoslovakia:  "Specific  training  is  recommended 

^German  Democratic  Republic:  " Training  courses  an*  orgam/iHl  hv  the 
Committees  in  cooperation  with  Akademie  fur  Arzlliche  Korthildung  der 
DDR." 

•^Federal  Republic  of  Germany:  "Fixperience  as  iv<iuired  by  la»  oi\  the 
control  of  communicable  diseases." 

•^Netherlands:  "The  scientists  should  he  trained  in  microhiologv 

^Norway:  The  Committee  certifies  training  and  I'xperlisi’  o(  personnel  *it 
adequafe." 

Sl'nited  Kingdom:  "Details  of  training  are  re(|uired.  the  emplosei  is  legally 
obliged  to  provide  suitable  training." 

^•United  States:  "Specific  training  not  required  How  eser  local  biohazards 
committees  are  required  to  certify  to  the  Nlll  that  the  training  and  e\|MTlise 
of  the  personnel  are  adequate." 


®Based  on  replies  from  previous  Questionnaires. 
^In  preparation. 


Appendix  lll-C— Information  on  International  Guidelines  for  Recombinant  DNA  • 325 


\lll.  Countries  in  which  the  guidelines  are  applicable 
only  to  biological  agents  containing  recombinant 
D\'A,  or  also  cover  the  recombinant  DNA  mole- 
cules themselves: 


Country 

Onl\'  to 
biological 
agents 

Also  recombinant 
DNA  molecules 

.Australia 

X 

Bulgaria 

X 

Canada  

(d 

P) 

Czechloslovakia  . . 

X 

Cierman 

Democratic 

Republic 

X 

f'ederal  Republic  of 

Germany 

X 

Finland 

X 

France  

X 

Japan  

X 

Netherlands 

X 

New  Zealand 

X 

Norway 

X 

South  .Africa 

X 

Sweden 

X 

Switzerland 

X 

Taiwan 

X 

United  Kingdom  . . 

X 

United  States  .... 

Xb 

U.S.S.R 

X 

^Guidelines  apply  to  all.  but  containment  is  not  required  for  naked  DNA. 
^ The  Guidelines  apply  to  recombinant  DN.A  e.'tperiments  that  are  not  ex- 
empt under  Section  l-E  of  the  Guidelines.  Recombinant  DN.A  molecules  that 
are  not  in  organisms  or  viruses  are  exempt  from  the  Guidelines  (I-E-1)." 

X/\'.  Groups/Committees  responsible  for  carrying  out 
monitoring  of  containment  procedures: 


Country  Group 

Australia Institutional  Biosafety  Commit- 

tees. 

Bulgaria National  Committee 

Canada "University  and  Medical  Re- 

search Council  Biohazards  Com- 
mittees” 

Czechoslovakia . . . "Under  consideration  of  the  Na- 
tional Institutes  of  Public  Health.” 

German 

Democratic 

Republic "Monitoring  is  carried  out  by 

local  Biosafety  Officers,  wbo  are 
representatives  of  the  Committee 
in  their  institutions.” 


Federal  Republic  of 

Germany Officers  for  Biological  Safety 

monitor  the  health  of  employees 


and  compliance  at  laboratories; 
ZKBS  (Zentrale  Kommission  fur 
die  biologische  Sicherheit)  has 
overall  responsibility. 

France "Local  safety  committees” 

Hungary "National  Institutes  of  Public 

Health” 

Japan "Principal  Investigator  and  Safety 

Officer” 

Netherlands "Site  Inspection  Commission” 

New  Zealand "Local  controlling  Committees 

are  charged  with  monitoring 
observance  of  Guidelines. 
Biological  Safety  Officers  are  ap- 


pointed to  take  immediate 
responsibility.” 


Norway "Physical  containment:  Norwe- 

gian National  Institute  of  Public 
Health.  Biological  containment: 
Committee.” 

South  Africa "Above  P3,  Biosafety  Committee 

of  Institute  involved  and 
SAGENE.  Below  P3,  SAGENE 
only.” 

Sweden Not  applicable. 

Switzerland "At  the  responsibility  of  either 

the  individual  investigator  or  a 
local  biohazards  committee.” 

Taiwan No  response 

United  Kingdom  . . The  Health  and  Safety  Executive 
United  States  ....  “Observance  of  containment  is  to 


be  monitored  by  biohazards  com- 
mittees located  in  institutions  in 
which  the  research  is  conducted. 
Effectiveness  of  containment 
procedures  is  to  be  monitored  by 
the  principal  investigator  who  is 
to  report  problems  to  tbe  NIH.” 

U.S.S.R "Local  biosafety  commission, 

State  Sanitary  Inspection  control 
group  of  Recombinant  DNA  Com- 
mission. 


326  • Impacts  of  Applied  Genetics — Micro-Organisms,  Plants,  and  Animals 


XV.  Countries  in  which  the  guidelines  apply  to  all  gene 
combinations  constructed  by  cell-free  methods,  or 
only  to  molecules  containing  combinations  of 
genes  from  different  species: 

Molecules 
All  gene  com-  containing 
binations  con-  combinations  of 
structed  by  cell-  genes  from 


Country 

free  methods 

different  species 

Australia 

X 

Canada  

X 

Czechoslovakia . . . 

X 

German 

Democratic 

Republic 

X 

Federal  Republic  of 

Germany 

x^ 

Finland 

X 

France  

X 

Japan  

X 

Netherlands 

x» 

New  Zealand 

X 

Norway 

X 

South  Africa 

X 

Sweden 

X 

Switzerland 

X 

Taiwan 

X 

United  Kingdom  . . 

Xe 

United  States  .... 

X 

U.S.S.R 

X 

^Federal  Republic  of  Germany  Self-cloning  experiments  involving  non- 
pathogenic  donors  and  hosts  shall  be  reported  to  ZKBS. 

^Netherlands  "The  definition  of  recombinant  DNA  has  recently  been 
modified  and  includes  the  insertion  of  chemically  synthesized  DNA  mole- 
cules into  a vector." 

^United  Kingdom  "The  Group's  provisional  interpretation  of  their  own  re- 
mit is  that  they  are  concerned  with  work  involving  genetic  manipulation, 
defined  for  these  purposes  as:  the  formation  of  new  combinations  of  her- 
itable materials  by  the  insertion  of  nucleic  acid  molecules,  produced  by 
whatever  means  outside  the  cell,  into  any  virus,  bacterial  plasmid,  or  other 
vector  system  so  as  to  allow  their  incorporation  into  a host  organism  in 
which  they  do  not  naturally  occur  but  in  which  they  are  capable  of  con- 
tinued propagation." 

XVI.^  Countries  in  which  the  guidelines  restrict  the  in- 
tentional dissemination  into  the  environment  of 
biological  agents  containing  recombinant  DNA: 


All  respondents  . . Yes^ 

Australia Not  explicity  so 

German 

Democratic 

Republic "Exceptions  have  to  be  discussed 

by  tbe  Committee  and  require 

special  permission  by  the  Minis- 
ter of  Health.” 


New  Zealand "Yes,  with  the  approval  of  the 

National  Committee." 

United  Kingdom  . . "The  question  has  not  arisen.” 


Other  respondents  No 

^Are  there  any  circumstances  under  which  such  dissemination  can  be  car- 
ried out? 

XVII.  Countries  in  which  the  guidelines  are  restricted 
to  recombinant  DNA  activities  or  also  cover 
other  areas  of  genetic  e;<perimentation: 

Recombinant  Other  areas  of 


Country 

DNA 

activities 

genetic 

experimentii 

Australia 

X‘> 

Bulgaria 

X 

Canada  

X'> 

Czechoslovakia . . . 

X 

German 

Democratic 

Republic 

X 

Federal  Republic  of 

Germany 

X 

Finland 

X 

France  

X 

Hungary 

X 

Japan  

X 

Netherlands 

X 

New  Zealand 

X- 

Norway 

X 

South  Africa 

X*' 

Sweden 

X 

Switzerland 

X 

United  Kingdom  . . 

X 

United  States  .... 

X 

U.S.S.R 

X 

^"Al  pn*sen!.  Ihe  lerms  ot  rrlrriMirr  o[  Ihr  \(.ulrni\  C oiiiinilti**- 
only  to  in  vitro  f*xpi*rim(»nls  li  v . itir  use  ol  n**»tri(  fion  rn/\  m«  « .in<l 
An  H(i  hoc  Academy  Committer  \s  .ihout  lo  investif(.ile  m \t\<>  uihmiU 
lion,  with  the  following  terms  of  i elerem  e 

1.  Kxamine  whether,  other  than  !)v  using  the  Iim  fmi(|ur  of  in  wfn»  »» 

combinanl  [)NA  construction,  new  hyhrid  nut  U*k  and  mo)n  iil«  ^ f>« 

produced  that  an*  potentially  dangertius  lo  human''  animals  oi  pUnu 

In  so  doing,  thi*  committee  should  gn  r pai  ticular  alleniHin  to  if><  f<  dh 
ing  possif)ilities: 

~ I he  use  of  mixed  infcM  lions  in\  oK  ing  human  or  aninul  \ w i > nr  ifu 
use  of  bacteria  or  fungi 

— rh(*  introdiKiion  of  foreign  D.NA  into  plants  and  thr  ptiNfuiti  -u 
new  plant  pathogens 

2.  ('onsider  whether  there  are  n’l  tain  i lasM‘s  of  \iral  paifu-*.*  t v . ^ 
polio)  on  whii'h  expi>rimentalion  should  not  he  < arnr<l  out  on  • ' « ')• 
cial  m*(‘d  is  de*monstrated 

*^  'work  with  animal  viruses  and  ( ells 

^'  i.f*.,  (*ell  fusion  will)  appnn  al  ol  National  ( onimilit**' 

^^  ’Olher  closely  relali'd  areas  are  also  * o\  i-r  rd 


Appendix  III  C— Information  on  International  Guidelines  for  Recombinant  DMA  • 327 


Will.  Countrivs  it}  which  the  rrcontbinant  DS'A  ad- 
visory committee  includes  public  representa- 
tives as  w ell  as  scientists: 


(~ountr\ 

Austialia 

Bulgaria 

Canaiia 

C'/t*th()sl()\  akia 
Dt'nmark 
(ii'rrnan 
l)f  mocratir 
Kepublic 

Fetleral  Kepuhlic  of 

(iermanv 

Finland 

FiMiH'e 

Hungary 

Italy  ' 

Japan 

Netherlands 
New  Zealand 

Norv\ay 

South  Africa 

Sw  eden 

Switzerland 

Faiw  an 

I'nited  Kingdom  . . 
I’nited  States  .... 

r.s.s.R 


\es ^ 

-\ 

X 

\ 

\ 

\ 


\ 

\ 

\ 

\ 

X 

X 

X 

X 

X 

X 

X 

X 

X 

X 

X 

X 

X 


Composition  of  D.N A adxisorv  committees  is  as  fol- 


lows: 

Australia 8 scientists 

Canada 5 laymen  ( 1 lawyer,  1 business- 

man, 3 generalists):  6 scientists  (2 
M.D.s,  3 \irologists/cancer  spe- 


cialists, 1 recombinant  DN',A  spe- 
cialist) 


Czechoslovakia ...  6 members  representing  molec- 
ular biology,  genetics,  microbiol- 
ogy-. medicine 

Denmark 9 scientists  and  administrative 

representathes. 


German 

Democratic 

Republic 3 geneticists,  1 biochemist,  2 bac- 

teriologists, 2 \ irologists,  1 jurist, 
1 representati\  e of  trade  union 
of  GDR. 


Federal  Republic  of 

Germany 4 experts  w orking  in  the  field  of 

recombinant  DXA  research;  4 ex- 
perts who,  though  not  w'orking 
in  the  field  of  recombinant  DNA 


Finland 

research,  possess  specific  knowl- 
edge in  the  implementation  of 
safety  measures  in  biological  re- 
search work,  particularly  how- 
ever in  microbiology,  cytobiolo- 
g\',  or  hygiene  and,  in  addition,  4 
outstanding  individuals,  for  ex- 
ample from  the  trade  unions,  in- 
dustry, and  the  research-promot- 
ing organizations. 

. . . 27  members:  6 molecular  biol- 

France  

ogy,  3 genetics,  3 microbiology,  1 
virology,  1 plant  physiology,  3 in- 
fectious diseases,  3 epidemiology, 

2 enteric  bacteria,  1 cell  cultures, 

3 public  health,  1 occupational 
health. 

. . . 13  members,  4 observers,  1 sec- 

Hungary 

retary 

. . . Scientists 

Italy 

. . . 8 molecular  biologists,  4 micro- 

Japan  

biologists,  1 civil  servant  (Health 
Ministry). 

. . . (Combines  both  Steering  Com- 

Netherlands  . . , 

mittee  and  Advisory  Group):  7 re- 
combinant DNA  scientists,  7 sci- 
entists in  other  fields,  6 special- 
ists in  medicine  and  biohazards,  2 
lawyers,  2 specialists  in  physical 
containment,  3 public  represent- 
atives. 

. . . 14  scientists  representing  genet- 

New' Zealand.  . . 

ics,  molecular  biology,  bacteriolo- 
gy, virology,  botany,  medicine, 
ethics  and  social  aspects  of  health 
and  health-care.  To  be  added:  a 
committee  composed  of  scientists 
and  representatives  of  industry 
and  trade  unions. 

. . 1 molecular  biologist,  1 microbial 

Norway 

geneticist,  1 virologist,  1 botanist 
(molecular  biologist),  1 human 
geneticist  (medically  qualified). 

. . 3 biochemists,  2 medicine,  1 vet- 

South  Africa  . . . 

erinary  medicine,  1 lawyer,  1 
artist. 

. . One  each  from:  Council  for  Sci- 

Sweden 

entific  and  Industrial  Research, 
Medical  Research  Council,  De- 
partment of  Health,  Department 
of  Agricultural  Technical  Serv- 
ices. Three  from  universities, 
public  and  legal  professions. 

. . No  response 

328  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Switzerland 


United  States  . . 


12  members  representing  medi- 
cine, microbiology,  molecular 
biology,  antibiotics,  industry, 
university  management,  and  7 
governmental  departmental 
assessors. 

Molecular  biology:  6,  Molecular 
Genetics:  5,  Ethics:  3,  Microbiol- 


ogy: 2,  Plant  Genetics:  2,  Law:  2, 
Environmental  Concerns,  Lab- 
oratory Technician,  Infectious 
Diseases,  Occupational  Health, 
Education:  1 each. 

U.S.S.R 8 scientists 

Yugoslavia 3 geneticists 


Appendix  IV 

Planning  Workshop  Participants; 
Other  Contractors  and  Contributors; 

and  Acknowledgments 


Planning  iforkshop  participants 

Philip  Bereano 
I'niversity  of  \\  ashington 
Ralph  Harciv 

E.  I Dll  F’ont  de  Nemours  & Co.,  Inc. 

Patricia  King 

Georgetown  l'ni\ersitv  Law  Center 
Charles  Lewis 

L'.S.  Department  of  .Agriculture 
Herman  Lewis 
.National  Science  Foundation 
Pamela  Lippe 
Friends  of  the  Earth 
\ ictor  McKusick 

Johns  Hopkins  University  .Medical  School 
EJena  ().  Nightingale 
Institute  of  Medicine 
Gilbert  Omenn 

Office  of  Science  and  Technolog\-  Policy 
Donna  Parratt 

Cogressional  Research  Service 
Walter  Shropshire 

Smithsonian  Radiation  Biologi'  Laboratory 
Leroy  Walters 
The  Kennedy  Institute 

Other  contractors  and  contributors 

Betsy  .Amin-.Arsala 
Richard  J.  .Auchus 

Massachusetts  Institute  of  Technologx' 

Fred  Bergmann 
National  Institutes  of  Health 
Charles  L.  Cooney 

Massachusetts  Institute  of  Technologx' 
Richard  Curtin 
Robert  A.  Cuzick 

Massachusetts  Institute  of  Technology 
Roslyn  Dauber 
■Arnold  L.  Demain 

Massachusetts  Institute  of  Technology 
Richard  B.  Emmitt 

F.  Eberstadt  &,  Co. 

Emanuel  Epstein 

Unh  ersity  of  California,  Davis 
Robert  F.  Fleischaker 
Massachusetts  Institute  of  Technology 


Odelia  Funke 
George  E.  Garrison 
F.  Eberstadt  & Co. 

Reinaldo  F.  Gomez 

iMassachusetts  Institute  of  Technology 
John  Hamilton 
Neal  F.  Jensen 
Scott  A.  King 
F.  Eberstadt  & Co. 

Harvey  Lodish 

Massachusetts  Institute  of  Technology 
L.  D.  Nyhart 

Massachusetts  Institute  of  Technology 
ChoKyun  Rha 

Massachusetts  Institute  of  Technology 
William  Scanlon 
■Andrew  Schmitz 

University  of  California,  Berkeley 
Michael  J^  Shodell 
The  Sterling-Hobe  Corp. 

David  Tse 

Massachusetts  Institute  of  Technology 
James  Welsh 
Montana  State  University 
George  Whiteside 

Massachusetts  Institute  of  Technology 
Bernard  Wolnak  and  Associates 
Nancy  Woods 
OTA  intern 

Acknowledgments 

A large  number  of  individuals  provided  valuable 
advice  and  assistance  to  OTA  during  this  assessment. 
In  particular,  we  would  like  to  thank  the  following 
people: 

John  Adams 

Pharmaceutical  Manufacturers  Association 
Rupert  Amann 
Colorado  State  University 
William  Amon,  Jr. 

Cetus  Corp. 

Daniel  Azarnoff 
Searle  Laboratories 
A.  L.  Barr 

West  Virginia  University 
K.  J.  Betteridge 

Animal  Diseases  Research  Institute 


329 


330  • Impacts  of  Applied  Genetics— Micro-Organisms,  Plants,  and  Animals 


Jerome  Birnbaum 

Merck,  Sharp  & Dohme  Research  Laboratories 
Gerald  Bjorge 

U.S.  Patent  and  Trademark  Office 
Hugh  Bollinger 
Plant  Resources  Institute 

G.  Eric  Bradford 
University  of  California 

Robert  Brackett 
Parke,  Davis  & Co. 

Robert  Byrnes 
Genentech,  Inc. 

Daniel  Callahan 
The  Hastings  Center 
Alexander  M.  Capron 

President’s  Commission  for  the  Study  of  Ethical 
Problems  in  Medicine  and  Biomedical  and 
Behavioral  Research 
Robert  Church 
University  of  Calgary 

H.  Wallis  Clark 
University  of  California 

Gail  Cooper 

Environmental  Protection  Agency 
Joseph  P.  Dailey 
Revlon  Health  Care  Group 
Frank  Dickinson 
Agricultural  Research  Center 
Donald  R.  Dunner 

Finnegan,  Henderson,  Farabow,  Garrett  &, 
Dunner 

Roger  B.  Dworkin 
Indiana  University  School  of  Law 
Richard  P.  Elander 
Bristol-Myers  Co. 

Peter  Elsden 

Colorado  State  University 
Haim  Erder 

University  of  Pennsylvania 
James  F.  Evans 

Pennsylvania  Embryo  Transfer  Service 
Kenneth  Evans 

Plant  Variety  Protection  Office 
Richard  Faust 
Hoffman-La  Roche,  Inc. 

Herman  Finke 
Sterling  Systems 
Robert  H.  Foote 
Cornell  University 
Orrie  M.  Friedman 
Collaborative  Research,  Inc. 

William  J.  Gartland,  Jr. 

National  Institutes  of  Health 
Kenneth  Goertzen 
Seed  Research,  Inc. 


Michael  Goldberg 
Food  and  Drug  Administration 
Maxwell  Gordon 
Bristol  Laboratories 
Lorance  L.  Greenlee 
Keil  and  Witherspoon 
Ralph  Hardy 

E.  I.  Du  Pont  de  Nemours  and  Co.,  Inc. 
W.  C.  D.  Hare 

Animal  Diseases  Research  Institute 
Paul  Harvey 

U.S.  Department  of  Agriculture 
Harold  W.  Hawk 
Agricultural  Research  Center 
Richard  L.  Hinman 
Pfizer,  Inc. 

Peter  Barton  Hutt 
Covington  & Burling 

E.  Keith  Inskeep 

West  Virginia  University 
Irving  Johnson 
Lilly  Research  Laboratories 
Charles  Kiddy 

Agricultural  Research  Center 
Thomas  D.  Kiley 
Genentech,  Inc. 

Carole  Kitti 

National  Science  Foundation 
Duane  C.  Kraemer 
Texas  A&.M  University 
Sheldon  Krimsky 
Tufts  University 
W.  W.  Lani[)eter 
I, elm  und  Versuchtsgut 
Earl  Lasley 

Monsanto  Farmers  Hybrid 

F.  Douglas  Lawrason 
Schering  Cor[). 

Bernard  Leese 

Plant  V'ariety  i’rotection  Office 
Stanley  Leiho 

Oak  Ridge  National  Laboratory 
Morris  Levin 

Environmental  Protection  Agency 
Herman  Lcnvis 
National  Science  Foundation 
Peter  Lihassi 

V'erner,  Lipfert,  Bernhard  N.  .\lacl’heiM)n 
Paul  J.  Luckern 
U.S.  Department  of  Justice 
Clement  Markert 
Yale  University 
Ralph  R.  Maun?r 

U.S.  Meat  Animal  Besearch  ( enter 


Appendix  IV— Planning  Workshop  Participants,  Other  Contractors  and  Contributors,  and  Acknowledgments  • 331 


Robert  McKinnell 
l'ni\ersitv  of  Minnesota 

K.ilw  arcl  Mearns,  Jr. 

( ast*  W estern  Reserve  l'ni\  ersity  School 
of  .Medicine 
Man  S .Michaels 
Stanford  I'niversity 
Kli/aheth  Milewski 
National  Institutes  of  Health 
Henry  I .Miller 

food  and  Drug  \dministration 
Paul  .Miller 
\V  R (irace 
A \ \alhando\ 

I'nixersity  of  Illinois 
(’laude  H.  Nash 

Smith  Kline  & I'rench  Laboratories 
IX)rothy  .Nelkin 
Resources  for  the  Future 
Gordon  Niswender 
(Colorado  State  I'niversity 
Klena  Dttolenghi-Nightengale 
National  Academy  of  .Medicine 
David  Padwa 
.Agrigenetics.  Inc. 

Seth  Pauker 

National  Institute  of  Occupational  Safety 
(!t  Health 
J.  B.  Peters 

V\  est  A'irginia  University 
Nancy  Pfund 

Stanford  University  School  of  Medicine 
James  Punch 
The  L'pjohn  Co. 

Neils  Reimers 
Stanford  I'niversity 
Ira  Ringler 
•Abbott  laboratories 
Roman  Saliwanchik 
The  L'pjohn  Co. 

Robert  B.  Samuels 
Beckman  Instruments 
George  E.  Seidel,  Jr. 

Colorado  State  L'niversity 
Sarah  .M.  Seidel 
Colorado  State  University 


Thomas  J.  Se.xton 
U S.  Department  of  Agriculture 
Brian  F.  Shea 
Ralph  Silber 
Stanford  University 
Elizabeth  L.  Singh 
Charles  G.  Smith 
Revlon  Health  Group 
•Animal  Diseases  Research  Institute 
Davor  Solter 

VVistar  Institute  of  Anatomy  and  Biology 
Mark  Sorrells 
Cornell  University 
G.  F.  Sprague 
University  of  Illinois 
Richard  Staples 
Cornell  University 
Gerald  G.  Still 

LIS.  Department  of  Agriculture 
Charles  VV.  Stuber 
North  Carolina  State  University 
Bernard  Talbot 
National  Institutes  of  Health 
Rene  Tegtmeyer 

U.S.  Patent  and  Trademark  Office 
Clair  E.  Terrill 

U.S.  Department  of  Agriculture 
Robin  Tervit 

Colorado  State  University 
Stephen  Turner 

Bethesda  Research  Laboratories,  Inc. 

L.  D.  Van  V'leck 
Cornell  University 

Robert  Walton 
VV.  R.  Grace 
Charles  Weiner 

Massachusetts  Institute  of  Technology 
Ray  W.  Wright,  Jr. 

Washington  State  University 
Susan  Wright 
University  of  Michigan 
Oskar  R.  Zaborsky 
National  Science  Foundation 

M.  S.  Zuber 
University  of  Missouri 


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