• TABLE OF CONTENTS
HIDE
 Front Cover
 Half Title
 Table of Contents
 Title Page
 Frontispiece
 Preface
 Acknowledgement
 Introduction
 United States Senate hearing
 An overview of biotechnology (Philip...
 Impact of biotechnology on agriculture:...
 Impact of contemporary biotechnology...
 Impact of biotechnology on biomedical...
 Impact of biotechnology on marine...
 Impact of bioengineering on biotechnology...
 A European perspective on biotechnology...
 The status and future of biotechnology...
 Biotechnology: the university perspective...
 Products and health care delivery...
 Changing role and scope of venture...
 Commercializing new products from...
 Commercializing biotechnology resources:...
 Tomorrow's science policy and biotechnology...
 Sustaining American leadership...
 Closing remarks (Kenneth R....
 Symposium program
 Back Cover






Group Title: Biotechnology: perspectives, policies and issues
Title: Biotechnology
CITATION PAGE IMAGE ZOOMABLE PAGE TEXT
Full Citation
STANDARD VIEW MARC VIEW
Permanent Link: http://ufdc.ufl.edu/UF00053779/00001
 Material Information
Title: Biotechnology perspectives, policies, and issues : an international symposium, held at the University of Florida, Gainesville, Florida, June 1-4, 1986
Physical Description: 247 p. : port. ; 24 cm.
Language: English
Creator: Vasil, I. K
University of Florida -- Institute of Food and Agricultural Sciences
Publisher: Published for the University of Florida Press by University Presses of Florida
Place of Publication: Gainesville
Publication Date: c1987
 Subjects
Subject: Biotechnology -- Congresses   ( lcsh )
Genre: government publication (state, provincial, terriorial, dependent)   ( marcgt )
bibliography   ( marcgt )
conference publication   ( marcgt )
non-fiction   ( marcgt )
 Notes
Bibliography: Includes bibliographies.
Statement of Responsibility: edited by Indra K. Vasil.
General Note: "Sponsored by the Institute of Food and Agricultural Sciences, University of Florida; the IC Institute, The University of Texas at Austin; and the RGK Foundation."
Funding: Florida Historical Agriculture and Rural Life
 Record Information
Bibliographic ID: UF00053779
Volume ID: VID00001
Source Institution: Marston Science Library, George A. Smathers Libraries, University of Florida
Holding Location: Florida Agricultural Experiment Station, Florida Cooperative Extension Service, Florida Department of Agriculture and Consumer Services, and the Engineering and Industrial Experiment Station; Institute for Food and Agricultural Services (IFAS), University of Florida
Rights Management: All rights reserved, Board of Trustees of the University of Florida
Resource Identifier: aleph - 001048460
oclc - 15791738
notis - AFD1505
lccn - 87014268 //r88
isbn - 0813008832

Table of Contents
    Front Cover
        Front Cover 1
        Front Cover 2
    Half Title
        Page i
        Page ii
    Table of Contents
        Page v
        Page vi
    Title Page
        Page iii
        Page iv
    Frontispiece
        Page vii
        Page viii
    Preface
        Page ix
        Page x
    Acknowledgement
        Page xi
        Page xii
    Introduction
        Page 1
        Page 2
        Page 3
        Page 4
        Page 5
        Page 6
        Page 7
        Page 8
        Page 9
        Page 10
        Page 11
        Page 12
    United States Senate hearing
        Page 13
        Page 14
        Page 15
        Page 16
        Page 17
        Page 18
        Page 19
        Page 20
        Page 21
        Page 22
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        Page 25
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        Page 27
        Page 28
        Page 29
        Page 30
        Page 31
        Page 32
        Page 33
        Page 34
    An overview of biotechnology (Philip H. Abelson)
        Page 35
        Page 36
        Page 37
        Page 38
        Page 39
        Page 40
        Page 41
        Page 42
        Page 43
        Page 44
        Page 45
        Page 46
        Page 47
        Page 48
        Page 49
        Page 50
    Impact of biotechnology on agriculture: plants (Peter R. day)
        Page 51
        Page 52
        Page 53
        Page 54
        Page 55
        Page 56
        Page 57
        Page 58
        Page 59
        Page 60
        Page 61
        Page 62
    Impact of contemporary biotechnology on animal science (Roger M. Weppelman)
        Page 63
        Page 64
        Page 65
        Page 66
        Page 67
        Page 68
        Page 69
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        Page 76
        Page 77
        Page 78
        Page 79
        Page 80
        Page 81
        Page 82
    Impact of biotechnology on biomedical science (Thomas W. O' Brien)
        Page 83
        Page 84
        Page 85
        Page 86
        Page 87
        Page 88
        Page 89
        Page 90
        Page 91
        Page 92
        Page 93
        Page 94
        Page 95
    Impact of biotechnology on marine and enviromental science (Rita R. Colwell)
        Page 96
        Page 97
        Page 98
        Page 99
        Page 100
        Page 101
        Page 102
        Page 103
    Impact of bioengineering on biotechnology (Alan Sherman Michaels)
        Page 104
        Page 105
        Page 106
        Page 107
        Page 108
        Page 109
        Page 110
        Page 111
        Page 112
        Page 113
        Page 114
        Page 115
        Page 116
    A European perspective on biotechnology (John Maddox)
        Page 117
        Page 118
        Page 119
        Page 120
        Page 121
        Page 122
        Page 123
        Page 124
        Page 125
        Page 126
    The status and future of biotechnology in developing countries (Yongyuth Yuthavong)
        Page 127
        Page 128
        Page 129
        Page 130
        Page 131
        Page 132
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        Page 137
        Page 138
        Page 139
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        Page 141
        Page 142
        Page 143
    Biotechnology: the university perspective (Robert Barker)
        Page 144
        Page 145
        Page 146
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        Page 148
        Page 149
        Page 150
        Page 151
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        Page 155
        Page 156
    Products and health care delivery (Gary R. Hooper)
        Page 157
        Page 158
        Page 159
        Page 160
        Page 161
        Page 162
        Page 163
        Page 164
        Page 165
        Page 166
    Changing role and scope of venture capital in the commercialization process of biotechnology (Scott A. Bailey)
        Page 167
        Page 168
        Page 169
        Page 170
        Page 171
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        Page 183
        Page 184
        Page 185
        Page 186
        Page 187
    Commercializing new products from plant biotechnology: problems and perspective (Cyrus McKell)
        Page 188
        Page 189
        Page 190
        Page 191
        Page 192
        Page 193
        Page 194
        Page 195
        Page 196
        Page 197
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        Page 200
        Page 201
        Page 202
        Page 203
        Page 204
        Page 205
        Page 206
    Commercializing biotechnology resources: competition and cooperation in global markets (George Kozmetsky)
        Page 207
        Page 208
        Page 209
        Page 210
        Page 211
        Page 212
        Page 213
        Page 214
        Page 215
        Page 216
        Page 217
        Page 218
        Page 219
        Page 220
        Page 221
    Tomorrow's science policy and biotechnology (Orville G. Bentley)
        Page 222
        Page 223
        Page 224
        Page 225
        Page 226
        Page 227
        Page 228
    Sustaining American leadership in biotechnology (Robert Rabin)
        Page 229
        Page 230
        Page 231
        Page 232
        Page 233
        Page 234
    Closing remarks (Kenneth R. Tefertiller)
        Page 235
        Page 236
    Symposium program
        Page 237
        Page 238
        Page 239
        Page 240
        Page 241
        Page 242
        Page 243
        Page 244
        Page 245
        Page 246
        Page 247
    Back Cover
        Page 248
Full Text










'I.














Major breakthroughs achieved in cellular and mo-
lecular biology during the last decade have made it
possible to modify plant and animal cells geneti-
cally. These powerful new techniques offer oppor-
tunities to improve utilization of microbes, animals,
and plants toward human welfare. Current scien-
tific and commercial developments in these areas
have raised important new policy issues concem-
ing the implications and commercialization of
biotechnology.
Participants in this symposium examined the state
of the art of biotechnology and focused on the
issues of university/industry relations, economic
opportunities, and ethical questions in the com-
mercialization of biotechnology and the trans-
ference of these technologies throughout the world.

The symposium provided a forum for exchange
among business, academia, and government on
present and future scientific developments and be-
yond to the implications and the commercialization
of biotechnology and related national and inter-
national policy issues.



~=^-^-






BIOTECHNOLOGY:
PERSPECTIVES,
POLICIES
AND ISSUES











CONTENTS


Page

Preface................... .................. ............... ix
Acknowledgments ......................................... xi


PART I

INTRODUCTION ............................................ 3
UNITED STATES SENATE HEARING........................ 15


PART II
ADVANCES IN BIOTECHNOLOGY:
BREAKTHROUGHS AND BOTTLENECKS

AN OVERVIEW OF BIOTECHNOLOGY
Philip H. Abelson ........................................... 37
IMPACTS OF BIOTECHNOLOGY ON AGRICULTURE: PLANTS
Peter R. Day .............................................. 51
IMPACTS OF CONTEMPORARY BIOTECHNOLOGY ON ANIMAL
SCIENCE
Roger M. Weppelman ...................................... 63


PART III
ADVANCES IN BIOTECHNOLOGY:
ENVIRONMENT, HEALTH AND ENERGY

IMPACTS OF BIOTECHNOLOGY ON BIOMEDICAL SCIENCES
Thomas W. O'Brien ........................................ 85
IMPACTS OF BIOTECHNOLOGY ON MARINE AND
ENVIRONMENTAL SCIENCES
Rita R. Colwell ......................................... 96
IMPACTS OF BIOENGINEERING ON BIOTECHNOLOGY
Alan Sherman Michaels ................... ................ 104











PART IV
GLOBAL, UNIVERSITY AND
INDUSTRY PERSPECTIVES
A EUROPEAN PERSPECTIVE ON BIOTECHNOLOGY
John Maddox ........................................... 119
THE STATUS AND FUTURE OF BIOTECHNOLOGY IN
DEVELOPING COUNTRIES
Yongyuth Yuthavong ....................................... 127
BIOTECHNOLOGY: THE UNIVERSITY PERSPECTIVE
Robert Barker ............... ......... ... ........... 144


PART V
COMMERCIALIZATION ISSUES
PRODUCTS AND HEALTH CARE DELIVERY
Gary R. Hooper ..................................... 159
CHANGING ROLE AND SCOPE OF VENTURE CAPITAL IN THE
COMMERCIALIZATION PROCESS OF BIOTECHNOLOGY
ScottA. Bailey ........................................... 167
COMMERCIALIZING NEW PRODUCTS FROM PLANT
BIOTECHNOLOGY: PROBLEMS AND PERSPECTIVES
Cyrus McKell ............................................ 188

PART VI
SPECIAL LECTURES
COMMERCIALIZING BIOTECHNOLOGY RESOURCES:
COMPETITION AND COOPERATION IN GLOBAL MARKETS
George Kozmetsky ......................................... 209
TOMORROW'S SCIENCE POLICY AND BIOTECHNOLOGY
Orville G. Bentley ......................................... 222
SUSTAINING AMERICAN LEADERSHIP IN BIOTECHNOLOGY
Robert Rabin ............................................ 229
CLOSING REMARKS
Kenneth R. Tefertiller ..................................... 235

APPENDIX
Symposium Program ......................................... 237








BIOTECHNOLOGY:
PERSPECTIVES,
POLICIES
AND ISSUES

An International Symposium
Held at the University of Florida
Gainesville, Florida
June 1-4, 1986

Edited by
Indra K. Vasil
Graduate Research Professor of Botany
University of Florida
Gainesville, Florida
Sponsored by the Institute of Food and Agricultural Sciences,
University of Florida; the IC2 Institute, The University of
Texas at Austin; and the RGK Foundation.
University Presses of Florida
University of Florida Press
Gainesville












UNIVERSITY PRESSES OF FLORIDA is the central agency for scholarly
publishing of the State of Florida's university system, producing books
selected for publication by the faculty editorial committees of Florida's
nine public universities: Florida A&M University (Tallahassee), Florida
Atlantic University (Boca Raton), Florida International University
(Miami), Florida State University (Tallahassee), University of Central
Florida (Orlando), University of Florida (Gainesville), University of North
Florida (Jacksonville), University of South Florida (Tampa), University of
West Florida (Pensacola).

ORDERS for books published by all member presses of University Presses of
Florida should be addressed to University Presses of Florida, 15 NW 15th
Street, Gainesville, FL 32603.

Copyright 1987 by the Board of Regents of Florida


Library of Congress Cataloging in Publication Data
Biotechnology.

1. Biotechnology-Congresses. I. Vasil, I. K. II. University of Florida. Institute of Food
and Agricultural Sciences.
TP248.14.B57 1987 303.4'83 87-14268
ISBN 0-8130-0883-2







































Francis Aloysius Wood
November 17, 1932-August 22, 1985


This volume is dedicated to the memory of the late
Dr. F. Aloysius (Al) Wood,
Dean for Research, Institute of Food & Agricultural Sciences,
University of Florida, Gainesville, Florida,
and
Chairman, Committee on Biotechnology,
National Association of State Universities and Land-Grant Colleges,
for his valuable contributions,
support, and understanding of the
value of biotechnology research and
development in agriculture.











PREFACE


The power, potential and promise of biotechnology has, in less than two
decades, stirred worldwide interest in the application of this technology
and its products for the improvement of plant, animal and human life.
Universities, industries, governments, foundations, and international or-
ganizations are creating new opportunities for research, development, and
applications of biotechnology. Early products of biotechnology, already
available in the market place, are directed toward human and animal
health. On the other hand, there are no major products of the agricultural
(plant) biotechnology which are commercially available, although it is
generally agreed that eventually the economic, social and political impacts
of agricultural biotechnology products will far exceed those of human/
animal biotechnology.
The euphoria of the early days of biotechnology, followed by serious
difficulties in achieving the objectives, has gradually led to formulation of
more realistic prospects and time tables. It has also become clear that
substantial new basic research, particularly in the biology of plants, will
be needed to exploit the full potential of these powerful techniques.
Even the current limited success of biotechnology has brought to atten-
tion important new concerns, problems and challenges. These generally
are not discussed at the scores of technical meetings held each year in the
United States and in other parts of the World. The perception of biotech-
nology in the public mind, and the concern for possible hazards to environ-
ment and life, have fueled considerable debate and discussion in commu-
nities, the Congress and the press. It was the late Dr. F. Aloysius (Al)
Wood, who suggested that an international symposium be organized to
focus attention on the many facets and problems of regulation, funding,
priorities, intellectual rights, industry/university relationships, commer-
cialization, societal and ethical concerns, international concerns and com-
petition, and training. A distinguished international group of individuals
representing academia, industry and governments was thus assembled to
discuss and explore the present status of biotechnology, and to offer a fo-
rum for the exchange of views among academia, industry and govern-
ment. I trust that the record of these discussions presented in this volume
will serve to bring much needed attention to, and encourage discussion of,
these varied and complex but critical aspects of biotechnology research
and development.
The international symposium on "Biotechnology: Perspectives, Policies











and Issues", was held June 1-4, 1986, at the University of Florida, Gaines-
ville, Florida. It included a United States Senate Hearing on Biotechnol-
ogy Commercialization by Senator Lawton Chiles (Democrat, Florida).
This proceedings volume includes most of the material presented in the
hearing as well as in the symposium (the complete program is included in
the Appendix).
In the preparation of these proceedings for publication I have benefited
greatly by the able and expert assistance of Lenie Breeze, who has cheer-
fully carried a heavy burden of work and provided the critical link be-
tween the authors, the press and myself. My gratitude also to Alicia Fry for
editorial assistance in the preparation of the proceedings.

Indra K. Vasil











Acknowledgments


Financial support provided by the following organizations for the inter-
national symposium "Biotechnology: Perspectives, Policies and Issues" is
gratefully acknowledged:
Institute of Food and Agricultural Sciences, University of Florida
The IC2 Institute, The University of Texas at Austin
The RGK Foundation, Austin
Agricultural Division, The Upjohn Company and Asgrow Seed Company
Deloitte Haskins & Sells
E. I. du Pont Nemours & Company
Gainesville Area Chamber of Commerce
Monsanto Company
Purina Mills, Inc.
Talquin Corporation (Progress Center)
Westinghouse Electric Corporation






PART I
INTRODUCTION













Welcome Address


Marshall M. Criser, Jr.
President
University of Florida
Gainesville, Florida
Welcome to Gainesville, the University of Florida and the international
symposium on "Biotechnology: Perspectives, Policies, and Issues." The
University is pleased to host this symposium which is designed as a forum
for leaders in business, government, and the academic world to look be-
yond research and scientific development to the implications and commer-
cialization of biotechnology. The symposium is also designed to look at the
related national policies on university and industrial relations, ethical
questions, and economic opportunities.
The University of Florida is one of the largest and most comprehensive
graduate research institutions in the nation. We are a land grant university
in a state with special needs and with soils and subtropical climate more
like other countries than other states.
The quality, quantity, and importance of the research conducted on this
campus has already been recognized by the Carnegie Commission and by
the Association of American Universities. The significance of the research
has also been recognized by the amount of funding received for research.
Here faculty in medicine, engineering, agriculture, pharmaceuticals,
physics, business, biology, computer sciences, and other areas are already
attracting more than $100 million a year in grants and contracts which
have helped develop Bioglass, the Kalman filter, Gencor, Gatorade, and
the first nuclear-pumped laser. From new varieties of peanuts and soy-
beans to toxin-synthesizing bacterial genes and synthetic insect hormones,
from new technologies in CAD/CAM and robotics to developments in ge-
netic engineering and breast cancer detection, University of Florida fac-
ulty are making major contributions to research.
What often begins as pure research is most likely to be conducted by the
faculty and funded by the federal government. Applied research may be
conducted by the faculty but is more likely to be funded by the private
sector. For all the contributions from research faculty, and for all the sup-
port from the federal government, the real application and impact occurs
when business and industry become involved.
One has to wonder what could be accomplished if the same emphasis
had been placed on biotechnology as has been placed on electronic tech-









M. Criser


nology since the first digital computer was invented by University of Flor-
ida alumnus John Atanasoff. Although we have seen the cloning of plants
from tissues, advances in cellular and molecular biology, embryo rescue
and cell fusion, gene transfer and other genetic engineering, biotechnology
has not received the same attention as electronic technology.
If we are to develop plants and animals that can withstand insects,
diseases, and adverse weather, while producing greater nutrition and other
products, we must dedicate the same kind of human and fiscal resources
that we have given to information storage, retrieval, manipulations, and
exchange.
The average American may be more concerned with losing weight than
having a new grain in his morning cereal, but the possibility of developing
public interest and support for biotechnology is probably better than ever
before in our history.
Following the famine in Africa that was so dramatically depicted in our
newspapers, on television, and through other media, public interest and
concern has been widely expressed-from an outpouring of individual do-
nations, to "Live Aid" and other mega events. In this country, the concern
for the starving in Africa has been translated into helping our own food
producers with "Farm Aid" and to helping our own hungry with "Hands
Across America."
In spite of the criticism that Americans think that they can solve world
hunger by sending a can of food to Ethiopia, most Americans and others
realize that education and agricultural development are the real solutions
to the long-range problems. It will not take much to convince Americans,
and others throughout the world, that biotechnology holds the greatest
promise for the future.
Perhaps we cannot develop a grain, or bean, or bird, or other food
sources that can grow in the desert. But we may be able to develop plants
to help stabilize the deserts or provide edible food sources from minute
water supplies. Is it unthinkable that a plant might be developed that
would pull hydrogen and oxygen from the air to provide its own source of
water?
Creating what was once the unthinkable is the real challenge of biotech-
nology. This is not just for Africa. In this country, with our seeming abun-
dance of food, we need to be concerned with producing food sources
which yield more per acre, require less pesticides, and less fertilizers. We
need crops that are more energy efficient and can be grown successfully
without potential damage to the environment.
Twenty-five years ago, it was unthinkable to have a calculator that
would operate on a battery smaller than a dime. It was unthinkable to








WELCOME ADDRESS


have a portable computer with 640K memory, and now we have portable
calculators and computers to help developments in biotechnology.
It took education, government, and business working together to
achieve the developments in electronic technology, and it will take the
same cooperation for advancements in biotechnology. John Atanasoff may
have been the academician who invented the digital computer, but it took
federal funding. And it still took IBM, Texas Instruments, Apple, and
many others to make that technology available to individuals like you and
me.
Nationally and internationally, we have a public interest that can be
channeled to support biotechnology. One of our very real tasks will be to
direct that interest, capitalize it, and use it for generations to come.
In Florida, the challenges of biotechnology have special meaning be-
cause our environment is different than in other parts of the country. The
soybean that grows well in Virginia, Ohio, or Idaho is not successful in
Florida. However, the varieties of soybeans and peanuts developed by our
faculty in the Institute of Food and Agricultural Sciences are very success-
ful here and in many parts of Africa and Latin America.
Florida's soil and climate have forced us to be concerned about pesti-
cides, plant nutrition, and plant tolerance to heat, drought, salinity, and
other factors. Our efforts to solve our own problems have led to interna-
tional studies in agriculture that have benefited other countries, as well as
our local farmers.
Because of our experiences and needs, because of the comprehensive
nature of our University, and because of the special dedication of our fac-
ulty, the University of Florida has a special commitment to biotechnology.
I am pleased that you are here for this symposium and to explore re-
search and scientific developments, their implications, and the related
concerns about university and industry relations, ethics, and economics.
The State of Florida is also very committed to biotechnology. I have
already mentioned some of the reasons, including our soil and climate.
Another reason, perhaps rather obvious, is money. Agriculture has always
been an important part of the state's economy, but economic growth in
Florida has continued to outpace the national economy, primarily because
of Florida's growth in high-technology manufacturing. Over the past dec-
ade, Florida has ranked seventh in the nation in terms of growth of manu-
facturing employment. Of Florida's top ten manufacturing firms, eight
are in high-tech.
Since 1975, two-thirds of all new manufacturing jobs in Florida have
been in high-tech manufacturing. Since 1975, high-tech employment has
more than doubled. The number of high-tech and high-tech service firms








6 M. Criser
has grown by 54 percent in the past five years. Today more than 27.3
percent of Florida's manufacturing jobs are in high-tech, compared with
18 percent nationwide.
While most of this high-tech growth has been related to machines, agri-
culture is still big business and the state recognizes the potential of biotech-
nology. Agriculture is still the state's second largest business, just behind
tourism. One of every five jobs is in agriculture. Each year, the impact of
agricultural production and processing on sales activities in Florida is
nearly $16 billion. With a ripple effect, the total impact is nearly $43
billion, including the wholesaling and retailing of food but excluding res-
taurants.
Combine the state's strengths in agriculture and high-tech, and you be-
gin to understand our enthusiasm for biotechnology.











Welcoming Remarks


Ray Iannucci
Executive Director
Florida High Technology and Industry Council
Office of the Governor
Tallahassee, Florida

Governor Bob Graham asked me to welcome you to Florida and to read
to you his remarks. I am here representing the governor.
The governor is honored that Dr. Kozmetsky and the University of Flori-
da's Institute of Food and Agricultural Sciences have chosen Florida as the
location for an international symposium on biotechnology. You are in an
exciting state and you are at an exciting university. Our state is committed
to forging new frontiers in science and technology. There is interest in our
industry, in our universities, and among our political leaders to support
research to help Florida become a national and international leader in
research.
Only a few decades ago what happened in Florida did not matter very
much. To most of America, we were small, poor, and far away. Today
what happens in Florida sets trends that effect the world. Florida is on the
move and the University of Florida is on the move, as are our other univer-
sities. Florida has cast off the reputation as a "sun and fun" state and
assumed its rightful position as a leader in innovation, higher education,
and research.
As a major economic development goal, the governor has targeted bio-
technology to be a major industry for Florida. This will require a solid
academic foundation, a world class research faculty, top quality facilities,
and instrumentation. Within our university system, the groundwork for
this foundation already exists.
The governor's vision is to build upon this foundation and develop Flor-
ida into an internationally recognized source at the cutting edge of re-
search in biotechnology. This will require commitment, tenacity, and fore-
sight. The challenge before us in Florida is to develop our case to the
legislature and how best to link the creative energies in the laboratories to
the marketplace.
The state must examine its support of basic research. Research partner-
ships must be formed. As the state provides the financing for research and
the scientists embark on a journey of discovery, the state policymakers









8 R. Iannucci

must develop a tolerance for ambiguity and willingness to see our universi-
ties create an inventory of knowledge from which material progress will
inevitably flow. An overwhelming majority of Florida's present industries
are life science based-health care maintenance, citrus farming, food
processing, fruit and vegetable farming, cattle and horse raising, forestry
and commercial fishing. This makes advances in biotechnology all the
more important to Florida's long-term economic well being, for while bio-
technology represents a new industry in its own right, it is also a valuable
resource of innovation that will help assure the future for many of Flori-
da's existing industries.
An investment on the part of this state toward the development of bio-
technology is an investment in keeping Florida competitive with the rest of
the world, and in preserving the existing economic base of this state.











Welcoming Remarks


Charles B. Reed
Chancellor
State University System of Florida
Tallahassee, Florida
This symposium is a serious occasion, and biotechnology is a serious
subject, but like all subjects in which there are elements of mystery, and
even some fear, this subject is one which has often inspired humor. As in,
"What do you get when you cross a saber-toothed tiger with a cobra? I
don't know, but I wouldn't want one admitted as an undergraduate-even
on a football scholarship:' Nor would I recommend letting it out of the lab
for a walk around the campus. And the reason I wouldn't goes to the heart
of our discussions here today. The opportunities we have today, and the
concerns of society about how we use those opportunities, are at the center
of our discussions in this conference.
I am not a bioscientist, but my lack of advanced training in the field
gives me one great advantage-the right to ask dumb questions. Those are
always the hardest to answer. The questions I pose in beginning this con-
ference are these: What do we mean by biotechnology? Is it just gene
splicing? Would it include naturally occurring bacteria which can clean up
an oil spill by digesting petroleum? Do we include the water purification
functions of the sawgrass of the Florida Everglades?
The definition I like, and which I commend to you, is that of the Office
of Science and Technology of the Congress. That definition is as follows:
Biotechnology includes any technique that uses living organisms (or parts
of organisms) to make or modify products, to improve plants or animals,
or to develop microorganisms for specific uses.
The strength of this definition is its inclusiveness, for it permits us to
come together from fields as diverse as molecular biology to that of practi-
cal agriculture, from laboratory work with DNA to the halls of our medi-
cal schools and hospitals, and from the realm of international scientific
cooperation to the role of finance and banking in encouraging and sharing
new technology.
In my brief words of welcome, let me offer one particular word of
encouragement. This is a field which is uniquely positioned for the devel-
opment of public/private partnerships, involving the institutions of the
State University System of Florida. As chancellor, I want to go on record









C. Reed


today as encouraging those partnerships, and promoting them. We need
them, and we are prepared to make them work in Florida.
This is a major, international symposium. I am honored to be here. I am
proud that our State University System, and in particular, our host institu-
tion, the University of Florida, have convened this symposium. And we
are all very gratified to participate jointly with the sponsors: the Institute
of Food and Agricultural Sciences; the IC2 Institute of The University of
Texas at Austin, Dr. George Kozmetsky, Director; and the RGK Founda-
tion.
This symposium has essentially three goals: (1) to examine the state of
the art of biotechnology; (2) to consider university and industry relations;
and (3) to review economic opportunities, ethical considerations, and
technology transfer issues related to the development and application of
biotechnology throughout the world.
Clearly, many-but not all-of the challenges and opportunities which
we face here today flow from our new-found ability to alter the genetic
material in plant and animal cells. It is a simple statement, but it has not
been a simple matter to acquire this new-found scientific competence.
For many thousands of years, the human race's collective body of
knowledge remained virtually static or grew at such a slow rate, that
adapting to change was not a great challenge. Today, coping with social
impacts of technological change is a virtual industry, with popular au-
thors, journalists, and therapists all finding their niche.
The fact is that we are simply creating new knowledge and a great deal
faster than we have ever done before. It is a measure of the velocity of
knowledge in our era that thousands of people around the world have lived
long enough to encompass the arrival of piloted flight, global communica-
tions, and the creation of weapons capable of ending all life-in just one
lifetime.
There are even a few people alive today who were born in the year
1884, the year in which Gregor Mendel, the founder of the science of
genetics, died. If that seems like a long time ago, it was. In the same year,
the French impressionist movement was in full flower, Brahms wrote his
third symphony, and Mark Twain published "Huckleberry Finn."
Yet only 102 years have elapsed since Mendel's death, and in that time,
human beings have learned how to elude barriers of time and space which
had constrained us from our first appearance on earth.
With those lessons, as in the case of every significant technology-in-
cluding the use of fire, the development of tools, and the ability to keep
close track of time-deep social impacts have accompanied the develop-
ment of new knowledge.









WELCOMING REMARKS


Just as the automobile, air conditioners, the airplane, the transistor,
telephone and television systems, and the microprocessor chip have
marked us and the society in which we live, so, too will biotechnology-in
ways we will consider essential and desirable, and in ways we will proba-
bly dislike. We will be marked by this technology, because this is a technol-
ogy which will be developed. We have no option of not going forward, for
if we stop, others will go forward-perhaps with less care, with less cau-
tion, less restraint, and less responsibility.
There are two fundamental reasons for this imperative of development.
First, it will be beneficial and profitable, and could provide one nation
with a critical advantage over all others. And second, the curiosity of the
human race does not allow it not to be developed. Our job is to see that this
new technology is developed responsibly, and used appropriately.
As researchers, business people, and citizens, we have an obligation to
do our work safely, and to produce a quality product. We must recognize
that we are part of society, and that we are indebted to society for provid-
ing us the tools with which to work. We must recognize that we will be
held accountable for the fruits of our work. It is possible that we will find
methods to feed the world and end hunger, protect our natural environ-
ment, cure disease, distribute prosperity widely throughout the world,
and it is possible that we will develop things which are harmful, even
dangerous.
Let me paraphrase a famous understatement of the significance of a new
technology. It goes like this, "Some recent work by leading researchers,
which has been communicated to me in manuscript, leads me to expect
that biotechnology may become a new and important branch of knowl-
edge in the immediate future. Certain aspects of the situation seem to call
for watchfulness and, if necessary, quick action on the part of the adminis-
tration."
All I have done is to substitute the word "biotechnology" for the word
"uranium." The author of this assessment was Albert Einstein. The docu-
ment was his famous handwritten letter to President Roosevelt. The date
was 1939. Let us recognize the potency of biotechnology as we contem-
plate its benefits. Let us assess the risks at every stage, and act accordingly.
May this major international forum serve as a signpost on the road to the
responsible development of this new technology.








UNITED STATES
SENATE HEARING










Excerpts from the
United States Senate Hearing
Conducted by the
Honorable Lawton M. Chiles

Testimony from Dr. David Glass
Vice President
BioTechnica International, Inc.
Boston, Massachusetts
Thank you for this opportunity to present an overview of issues impor-
tant to the commercialization of biotechnology. In presenting this over-
view, I would like to approach it from two different perspectives. The first
is from the perspective of what the federal government can do to help
industry with some of the issues and problems that I and the other speakers
will be discussing today. As we will see, many of these issues are govern-
mental in nature, so it is appropriate for there to be some governmental
role in the responses to these issues.
The second perspective I would like to bring is that of a small company.
You have just noted the importance of small biotechnology companies in
this country in the commercialization of biotechnology products and, very
often, we in the small companies have a slightly different perspective on
things than some of the larger companies who are working in this field. My
company, BioTechnica International, is such a small company. We were
founded five years ago in Cambridge, Massachusetts, and we are a genetic
engineering research company developing products primarily in two ar-
eas-agriculture and dental diagnostics, diagnostic kits for the detection of
human periodontal disease and other dental diseases.
Our other interests in agriculture lie primarily in two areas-the devel-
opment of improved microbial soil inoculants, such as Rhizobium, and the
development of improved crop varieties themselves, primarily through the
use of recombinant DNA techniques. Our first products here will largely
involve herbicide-tolerant varieties of important crop plants. So I hope in
coming from a small company, I can provide the small company perspec-
tive in my remarks this morning.
Although I, and other people, talk about biotechnology as an industry, it
really isn't an industry so much as it is a collection of research tools and
techniques, most of which having been developed in the past 10 or 15









D. Glass


years, and all of which promise to revolutionize the way living organisms
are used for industrial purposes. Particularly, we are all aware of the great
commercial benefit and the great benefit to society that biotechnology is
expected to have in a number of industries, particularly human and ani-
mal health care, crop agriculture, and other areas such as food processing,
chemical processing, and energy generation.
Most of these research tools were invented here in the United States
within the past 10 or 15 years, stemming from basic research done in the
1970s at the nation's universities, largely funded by federal research
grants, and indeed, it was this large influx of federal research funding
which was largely responsible for the United States holding the clear un-
contested lead in biotechnology as the 1970s ended, and that is both in the
basic research area and in the emerging commercialization of biotechnol-
ogy that began around the end of the 1970s decade. However, as we have
moved from the seventies to the eighties and the technology has begun to
move to the marketplace, the United States has seen its lead slowly eroding
to the point today where we can say that we only hold a fragile lead over
our international competitors. This is the first issue that I would like to
raise today, the issue of international competitiveness and how the United
States might maintain its lead. While this subject is often mentioned by
industrial speakers in settings such as these, it is difficult to over-exaggerate
the importance of this issue for the industry and for the country. We cannot
let happen to biotechnology what has happened, as you noted to so many
other areas of technology in the past 20 years, areas that were largely
invented in the United States, but would end up being primarily commer-
cialized by companies in other countries.
I would like to mention two particular manifestations of this, one a
specific case and one a general case, where present governmental policies
either are, or run the danger of, contributing to this erosion of the United
States' lead in the technology.
A specific example is the situation regarding the export of unapproved
pharmaceutical products. Current United States law prohibits a company
to export a pharmaceutical product before it has been approved by the
Food and Drug Administration. Because the FDA review procedures are
so thorough and so lengthy, most drug products receive approvals in for-
eign countries before they receive FDA approval here in the United States.
Companies not wishing to lose a year or two or more of marketing time
will find a way to manufacture these products outside of the country, so
that they can supply markets where they receive regulatory approval.
When this is done, American jobs are lost and American technology is
transferred overseas, neither of which are particularly happy occurrences.









EXCERPTS FROM THE UNITED STATES SENATE HEARING


I might add that this circumstance affects biotechnology companies in
particular because most small biotechnology companies need larger corpo-
rate partners to market pharmaceutical products, and this state of affairs
would prompt them to look for foreign collaborators rather than collabo-
rators in the United States.
As I am sure you know, there have been several bills introduced into
Congress to alleviate this situation, one of which was recently passed in the
Senate about a month ago. This bill would allow certain pharmaceutical
products to be exported to certain countries once they have received regu-
latory approval from those countries. It is very important that this bill be
passed in the House and signed by the President. It is a very important bill
for that sector of the biotechnology community developing products in the
pharmaceutical area. Not only is that the largest sector of the biotechnol-
ogy industry, but it also could well be the one promising the most benefits
to society in terms of novel therapeutic products that treat cancer and
other deadly diseases.
A more general problem regarding U.S. competitiveness is the overall
regulatory structure and climate in the United States. The United States,
of course, is generally known to have perhaps the toughest regulatory re-
gime in the world. Now oftentimes this is absolutely necessary for the
adequate protection of the public health and safety and for the protection
of the environment, but it is important to note that when overly strict
regulations are applied to an emerging technology at an early stage in that
technology's growth, and in addition, at early stages of product develop-
ment, you run the real risk of stifling the development of the industry. I
think this will be most keenly felt in the areas of agricultural biotechnol-
ogy, as I will describe in a moment.
In general, I would like to point to the regulatory climate and the regu-
latory regime as the second major area of importance to the commerciali-
zation of biotechnology, and of course, a speaker later this morning, Mr.
Korwek, will go into this issue in more detail.
As I have said, this situation will primarily affect products in agricul-
tural biotechnology, and this is primarily because of the way that agricul-
tural products are developed in this country. That is not just biotechnology
products, but agricultural products in general. Companies developing ag-
ricultural products such as a new pesticide or a new strain of crop plant
need to do intensive testing on an iterative basis in the field to evaluate
what sometimes are many potential product candidates to narrow it down
to the one or two that are going to be commercially viable. Greenhouse
testing is not a 100 percent accurate indication of what is going to happen
in the field. This is the reason why companies have to do extensive field









D. Glass


testing. As I have said, in an iterative type of way, you take what you learn
the first year and then go back to the field the second year and so on,
repeating the process until you have come down to the candidate that you
know will work. What this implies, at least in biotechnology, is that many
of these early field tests that have been lately proposed and have been the
subject of some controversy, are actually highly preliminary product pro-
totypes. Some of them aren't even product prototypes at all, as much as
they are test cases or preliminary products, so by imposing stringent regu-
latory requirements for field testing on these types of tests runs the risk of
hindering innovation and stifling of development of agricultural products
from biotechnology. This, of course, is not to say that there shouldn't be
any regulation of field testing, and indeed, one of the challenges that faces
us is to come up with a regulatory regime for field testing that will allow
products to be field tested and eventually to allow products to come to the
market while still protecting the environment, and while still insuring the
public that health and safety considerations will be observed.
Currently in Washington, we have what amounts to a logjam right now
with the government not quite knowing how to regulate these initial field
tests. The result being that field tests, at least of microorganisms, are not
getting done. A colleague of mine, Dr. Ralph Hardy, has made a proposal
for a potential solution to this logjam which I would like to briefly mention
here today for possible discussion later on. He has proposed that the fed-
eral government establish publicly-owned, professionally-managed field
testing sites perhaps utilizing existing agricultural testing stations run by
the government. At these stations, companies would come using their own
personnel to conduct the tests, but they would be overseen by personnel
from the public sector who have the responsibility for running the field
site, of course, but also for overseeing the test. In this way, the government
and the public can be assured the tests are done according to the proper
protocols and with the proper monitoring, while still allowing the test to
go on in a timely basis.
Let me close off talking about regulations by simply noting, as I did
earlier, that we are not saying that there should be no regulations. Every-
one in industry believes that there should be regulations in biotechnology,
particularly in the environmental area. All that industry is asking is a
stable, predictable regulatory regime by which we will know in advance
what we need to do to get our products tested and out to the market, what
tests we need to do, how long those tests will take, and what it is going to
cost. Knowing this, we can incorporate all this information into our strate-
gic planning and this will be a desirable result for the industry. I think








EXCERPTS FROM THE UNITED STATES SENATE HEARING


clearly the administration and the Congress are moving towards this, but
sometimes the pace of this movement seems to be a little slow.
Let me move on to some other areas that are of importance to the com-
mercialization of biotechnology. Our next speaker, Congressman MacKay,
will be talking about issues of intellectual property protection. This is
clearly of great importance to the industry because no company will com-
mit the large amounts of time and money to develop a product without
knowing that they will have adequate protection for their products under
the nation's intellectual property laws, primarily the patent system. The
biotechnology industry has been heartened in recent years by two deci-
sions, one the famous 1980 Chakrabarty decision of the Supreme Court
which held that microorganisms could be patented, and more recently the
decision by the Patent Office Board of Appeals in the Hibberd case about
six months ago that plant materials can be patented under the general
patent statutes. They are encouraging and lead us to believe that we will
be able to have adequate patent protection for our products and biotech-
nological inventions; however, let me insert the caveat that until there is
more experience with biotechnology patents in the patent office and in the
courts, more case law, perhaps even more litigation, it will not be clear to
what extent biotechnology patents will be enforceable, what scope will be
available for them, and frankly, whether or not patents will have any
value in such a rapidly moving field of technology as biotechnology. How-
ever, right now we are all placing a lot of stock in the patent system. We
are all spending a lot of money trying to patent our inventions, and for this
reason, the U.S. Patent Office needs to be strong and needs to be able to
deal with these applications as they come in. As I am sure you know, there
is now a considerable backlog of patent applications at the Patent Office,
currently running about a year-and-a-half between initial application and
first office examination, with issuance a year or two after that. The Patent
Office needs more examiners and they need better-trained examiners in the
area of biological sciences. This should also be a responsibility of the gov-
ernment to make sure that the Patent Office is well-equipped to deal with
these issues.
Let me say a few words about potential legislative approaches to the
patent system, knowing that Congressman MacKay will probably spend
some time dealing with these issues. There have been a couple of potential
new areas of legislation that have been mentioned and proposed in Con-
gress. I will say a few words about two in particular. The first is patent
term restoration, whereby a company is compensated for lost marketing
time under its patent while it is waiting for marketing approval from a
federal regulatory agency. Patent term restoration for certain pharmaceu-









D. Glass


tical products was passed two years ago by Congress and there are now
several bills pending that would apply the same type of patent term resto-
ration to agricultural products. Those of us in the agricultural sector
would welcome agricultural patent term restoration.
Another major issue is process patent infringement. This is a situation
where a company holds a U.S. patent to manufacture a product, but does
not hold foreign patents on that process. A competitor infringes the patent
overseas and then comes back to the United States to sell the product.
Current law gives the U.S. company very little recourse against such in-
fringement. There are several laws pending in Congress that would make
this act an infringement, just as if the infringement occurred in the United
States. This type of bill is extremely important for the biotechnology com-
munity, particularly because so many of our patents will be process patents
covering manufacturing processes, rather than product patents, so this too
is a piece of legislation that we would like to see passed.
Another area that is of great importance to the biotechnology industry,
as well I am sure to a great many people in the audience today, is the issue
of basic research and the continued funding for university programs in
molecular biology, genetics, and other related areas. We have already
noted that the biotechnology industry owes its start and owes a great debt
to the large amounts of federal funding and the excellent basic research
conducted in this country in the 1970s. We all continue to be dependent on
future research at American universities as the industry develops. By this I
mean not only basic research by which we learn more about life and life
processes, but also the kinds of applied research sometimes funded by cor-
porations, sometimes funded by the government, which actually lead to
potential product opportunities for small companies and large companies
alike. We are also dependent on a third area of university research which
has become increasingly important in today's regulatory climate, and that
is research in the areas of risk-assessment, particularly environmental or
ecological research which can and will be used to support the kinds of risk-
assessments the companies will need to provide support for the registration
and approval of their products down the line.
All these types of research are important to be done at universities. Some
of the larger companies in our business, the established companies, are of
course, funding a great deal of research and will continue to do so. How-
ever, funding significant amounts of research is far beyond the abilities of
companies such as my own, and I think it is safe to say in this country that
funding of basic research remains the responsibility of the government.
For the biotechnology industry, it remains a very important responsibility
and function of the federal government.









EXCERPTS FROM THE UNITED STATES SENATE HEARING


Let me also just mention that in general, the relationships between in-
dustry and the universities in this country remain important. I believe
some of our later speakers will discuss in more detail some of these issues
concerning such relationships, including licensing agreements and com-
mercial arrangements. We are all dependent on the universities for the
training of our future employees, the scientists, engineers, and managers,
that we will need as our companies grow and develop. Again, I think we
will hear more on this topic from our speakers later this morning.
Another issue of great importance is the general issue of economic Oevel-
opment. That is, how can the government help us to foster a fa, able
climate for the development of biotechnology companies? Let me just say
two brief words about this. In general, one thing that would be very help-
ful would be governmental programs that would help foster a favorable
climate, economic and otherwise, for the development of companies and
for the expansion of companies. In this country, we have seen models in
certain state governments setting up biotechnology programs or biotech-
nology centers designed to provide economic incentives, tax breaks, etc.,
for companies to relocate in these states, expand their facilities into these
states, or set up manufacturing facilities. We have also seen foreign coun-
tries setting up the same types of programs. Particularly some of the Euro-
pean countries are establishing the same sorts of programs and are very
aggressively contacting U.S. companies, primarily small biotechnology
companies, to set up their manufacturing facilities overseas. This of
course, gets back to the international issues I mentioned earlier. It is in the
best interests of the U.S. government not to allow foreign countries to
entice our companies away from us, so the federal government should
consider what types of programs it might be able to do along the lines that
some of our own states have been doing.
The other issue in the area of economic development and capital forma-
tion is what types of financial incentives are appropriate for an industry
such as biotechnology. Here I will only note that the more traditional
means of financial incentives, such as R&D tax credits, are not particularly
useful for companies such as ours who are not paying taxes because we are
not making a profit yet, so the more traditional areas of financial incen-
tives might not be as applicable to our industry as it might be to other
industries. Industry and government together should work towards find-
ing creative, innovative ways to provide incentives for the types of longer
term, higher risk ventures that are characteristic of the biotechnology in-
dustry.
Let me close by discussing one last issue that, in the end, will probably
prove to be more important than all these other issues combined, and that









D. Glass


is the issue of public acceptance of the biotechnology industry and of our
products and of our activities. In many cases, the general public will be the
ultimate consumer of biotechnology products. In other instances, and re-
ally in all instances, the public, through their elected representatives and
through the appointed members of the federal government, have the re-
sponsibility for ensuring the safety of our products and assuring that every-
thing we do is done according to proper procedures. Clearly, there is a lot
to be done in the area of public information, leading toward public ac-
ceptance of what biotechnology does. Several areas are worth pointing out
here.- Certain sectors of the public still have religious or moral concerns
about what we do and about the effects of biotechnology. They will need
to be addressed in the years to come. In addition, there are a whole host of
ethical issues concerning the way the research is carried out, the impact of
the research and its products on the economy, and many other factors. All
of these need to be considered.
Then finally, there are the health and safety concerns and the environ-
mental safety concerns which today are very prevalent in the public, and
which, of course, are of extreme importance as the industry moves from
the laboratory to the marketplace, and particularly as the industry moves
from the laboratory to the field in agricultural field testing.
The solution to all these problems must come through considerable ef-
forts of public education and public information dissemination. This
largely has to come from the industry itself, as we all have to be much
more forthcoming with the public, with the trade press and the lay press,
the popular press, the Congress, and with other members of the public
sector. Industry is committed to doing this and, through our trade associa-
tions, have lately begun such public information activities. But we do ask
that wherever appropriate, and whenever possible, the government and
other members of the public sector help us in this regard, particularly from
the university sector, where a few words from a qualified academic expert
can sometimes be very useful in putting things into a proper perspective.
I hope that in this brief time I have been able to provide an overview of
important issues for commercialization of biotechnology. I am sure that
many of the points I have discussed will be discussed in much greater detail
by the speakers and witnesses who follow. I will be glad, Senator, to an-
swer any questions that you might have either now or later.











Testimony by Dr. Jefferey Burkhardt
University of Florida

I appreciate the opportunity this morning to address you on the issue of
ethics, social values, and the new biotechnologies. The new biotechnolo-
gies, unlike many other techniques or technologies, present us with the
opportunity to assess our values and scientific and institutional commit-
ments before we have plunged head-long into adoption, diffusion, and
commercialization.
Many mechanical technologies are already in place. We are in a position
to assess their impacts, but only after they have occurred. We know, for
example, that the mechanical harvester has had profound social and eco-
nomical effects. Some of these have been for good, some for ill. The
present state of biotechnology allows us to reflect on possible effects before
they occur. Hopefully we might prevent many, if not all, ill effects from
occurring.
The new biotechnologies also present us with the opportunity to reflect
on what our values are-that is, what we as individuals and as a society
find really important. What are our values regarding science, technology,
our institutions, and the environment? What value conflicts do these tech-
nologies present us with or exacerbate? What problems might they present
us with in the future?
My comments today are based on a study I conducted from 1983 to 1985
concerning the ethical and value issues surrounding the development and
applications of the new biotechnologies in agriculture, particularly re-
garding plant improvement. The study was funded by the National Sci-
ence Foundation's Ethics and Values and Science and Technology Pro-
gram. The other investigators and I personally interviewed over 100
scientists and research administrators across the country in both the public
and private sectors. We sought to ascertain exactly what was being done
with biotechnology in plant improvement, as well as what impacts bio-
technology is having and will have on the practice of science and on the
broader society.
My role as ethicist was to frame questions concerning values, interpret
and clarify the answers received from interviewees, and most importantly,
draw from our interviews the logical implications regarding value com-
mitments, institutional changes, and future prospects for biotechnology. I
would like to briefly summarize concerns voiced by scientists I inter-
viewed, as well as some of the implications of these concerns.
There are three distinct, though interconnected, sets of ethical issues
which can be gleaned from discussions with those in biotechnology. These










J. Burkhardt


three ethical problem areas are professional ethics, the changing mission of
public institutions, and responsibilities regarding the environment, future
generations, and the future of science.
The professional ethics problem area includes issues related to the
proper relationship between the practice of science, professional responsi-
bility, and personal integrity. Some of the cases that were raised as being of
major importance to bench scientists were the following. Biotechnology is
"in." Since the passage of the Plant Variety Protection Act, and since the
1980 Chackrabarty decision, plant varieties and novel life forms can be
patented. There is, understandably, a race among scientists to be the first
to introduce a bioengineered product to the public, or in the case of the
private sector, being the first to develop a patentable product for the mar-
ket. This can conceivably cause sloppy science, even dangerous science.
The concern is that long-standing cannons of professional responsibility,
including commitment to careful, rigorous, checked and re-checked scien-
tific results, might give way to fast and loose science. In the long-run,
shoddy science or faulty products would undoubtedly be excised from the
scientific community or from the market. The concern is, however, about
the short-run.
New university/industry relations, such as privately funded biotech in-
stitutes on university campuses, also cause concern. Proprietary rights,
which Congressman MacKay spoke about this morning, of corporate fund-
ing sources to biotechnologies can conceivably impede the free flow of
scientific information, even among public sector researchers, thereby de-
creasing the rate of discovery, replication of results, and so forth. Again
and again, scientists I spoke with voiced difficulties with sharing informa-
tion. If scientists can't legally communicate with each other, progress in
science is hampered.
The potential for conflicts of interest was also repeatedly noted. Scien-
tists and administrators have expressed concern that as private funds are
increasingly available, while individual public sector scientists retain a
university component in salary or research support, situations where the
scientist is serving two masters may arise. More importantly, however, is
the potential for scientists to use information or research results for per-
sonal gain, whatever the funding source. Both scientists and administra-
tors know the danger of violations of employment contracts and role re-
sponsibilities. Certainly the potential for conflicts of interest or contractual
violations existed long before the emergence of biotechnology; however,
the sheer amounts of monies to be made from biotechnological products
and processes have increased the opportunities for such conflicts.
The National Academy of Sciences and the National Association of State









EXCERPTS FROM THE UNITED STATES SENATE HEARING


Universities and Land Grant Colleges have attempted to prevent such
problems through legal mechanisms, making as specific as possible the
nature of public/private agreements, detailing what all parties can expect.
How individual scientists or administrators follow the letter or the spirit of
the law remains to be seen. Even with some legal instruments available or
in place, those involved in biotechnology continue to express concern
about honesty and conflicts. Some go so far as to blame the potential for
dishonesty or conflicts on the system. This leads me to the second problem
area.
The administrators I talked with expressed concerns about ethics and
values in their institutions. Among the situations cited were the following.
Funding cut-backs for public research or at least greater competition for
scarce resources have led public research administrators to seek private
assistance and also to look to shorter term results for R&D projects. Private
firms have looked to the university as a source of scientific talent with labs
and research assistants already there. Many administrators raise the ques-
tion of whether new institutional arrangements such as university/industry
contracts or privately supported biotech institutes on university campuses
may too drastically shift the historical division of labor between public and
private research agendas, perhaps threatening the kinds of research tradi-
tionally done in the public sector.
Questions were raised about the public sector's ability to serve a broader
constituency such as small farmers, farm labor, backyard gardeners and
the like. While most scientists and administrators believe that these tradi-
tional groups will be served over the long-term through biotechnology, the
short-term effects are what worry them. Interestingly, many private re-
search administrators express similar worries.
One specific problem raised in regard to the public sector was that some
programs or departments which, because of their discipline, cannot re-
ceive research support related to biotechnology might suffer. Among them,
for instance, social sciences, such as rural sociology and agricultural eco-
nomics, and perhaps even agricultural engineering. Since these programs
or departments have historically served a broad-based constituency, in-
cluding rural communities and small farms, de-emphasis of these pro-
grams in the overall research agenda may have negative social effects.
The issue here is the mission of public research. Most of the scientists I
talked with, and administrators as well, recognize that the mission of a
public institution will change as client groups change. In Florida, for ex-
ample, the rural/urban shift means food and agricultural research must
now pay closer attention to the needs of urbanites. Biotechnology can assist
them in development of techniques and products serving urbanites. How-









26 J. Burkhardt
ever, in the minds of many scientists, administrators, and client groups,
this research should not lead the way toward the more rapid decline in
rural clientele by undercutting research favoring small farms or rural com-
munities.
The last problem area is more amorphous, but it was raised indepen-
dently by scientists and administrators that I talked with. The last issue is
what we will leave posterity. There is an overarching sense that with any
new scientific discovery, potential exists for tremendous beneficial effects.
There is also a sense that ill effects are also possible. Most people were
aware of Rifkin's lawsuits raising the spectra of the potential long-term
environmental harms which could result from the release of bioengineered
organisms into the environment. Many felt that whatever the merits of
such cases, environmental effects cannot be lightly regarded. But interest-
ingly, part of the problem people noted was that neither the public, nor
those involved in biotechnology, were fully aware of the risks or lack of
them associated with biotechnology. To my mind, this raises a question
about the extent to which those involved in risk-assessment or decision-
making about biotech might be ethically obligated to inform the public
and others about those risks or lack of them. If we have adequate risk-
assessment, then, let us inform those concerned about biotechnology so as
to allow us to continue sound research in this area. If we don't have ade-
quate risk-assessment, more or better is probably necessary.
One final issue was raised. This is the issue of what sort of science we
will leave future generations. The new biotechnologies are the result of the
long-standing historical trend toward mechanistic reductionism, the re-
view that all phenomena can be explained only in terms of the complex
working of increasingly smaller and smaller parts. No one has disputed,
nor can dispute, the profound successes that this philosophy of science has
had across the board in terms of products, processes, and the amelioration
of the human condition. Nevertheless, concern was raised. If we become so
enamored with biotechnology as to systematically ignore more holistic sci-
ences or methodologies, or if we commit resources in such a way as to
destroy the base for broader research, or if we nearsightedly come to view
all facets of human life only in terms of the most microbiological processes,
we may be doing posterity a disservice. We may be blatantly violating
ethical obligations to the future.
I have spoken on three problem areas where ethical and value issues
have, and will, continue to emerge in regard to biotechnology. In my judg-
ment, this implies that further research is necessary on the ethical and
value dimensions of biotechnology, not only in agricultural resources, but
also in medicine, pharmaceuticals, and the like. A more complete aware-









EXCERPTS FROM THE UNITED STATES SENATE HEARING


ness of what we as individuals and we as a society think is important, and
what conflicts among various value systems exist, can only aid in decision-
making. Individual research by philosophers and social scientists, but
more importantly, interdisciplinary and joint research is clearly implied.
Every sponsor of biotechnology research and development projects should
perhaps have a values-or ethics component built in. Many of the risks or
impacts associated with biotech are simply not known at this time, but
without broad and forward-looking perspectives introduced into the very
core of our research, we may come to know only too late the full conse-
quences.
Even with the proliferation of research on ethical and value implica-
tions, additional actions may be necessary. Education for values awareness
is another need. Decisions are made by individuals. Many ethical and
value problems are undoubtedly resolvable in the consciences of individual
decision-makers. The presumption is, however, that scientists and deci-
sion-makers know how to see and how to address an ethical or value prob-
lem when they fall upon one. Here at the University of Florida and at
universities across the country, we have begun to address the mistake in
this presumption. Courses are offered where future scientists and decision-
makers can explore ethical problems they will face in their professional
lives. In these courses, future leaders are provided with logical, concep-
tual, and analytical skills appropriate to perceiving, facing up to, and
resolving where possible, ethical issues such as conflict of interest, scien-
tific honesty, and the goals of our institutions, including the future of sci-
ence.
Despite their importance, additional research and college courses in sci-
entific and professional ethics are not enough. There are thousands of sci-
entists and administrators who cannot wait for future research results or
for present students to join the ranks. The value and ethical issues involv-
ing biotechnology are presently here. This is why presently existing means
may be used to draw attention to and help address ethics-in colleges of
agriculture, in particular.
Extension may be the vehicle for providing this necessary service. Exten-
sion programs focusing on professional ethics for private sector scientists,
for example, may help fill the gap in ethics awareness. Programs on mana-
gerial ethics for public and private administrators might serve a similar
purpose. In general, values and ethics awareness programs are necessary
both to explore, as well as to explain, the benefits and risks of biotechnol-
ogy. These would enable more scientists and administrators to better assess
the complete aspects of biotechnology.
Others here today are, of course, better qualified than I am to speak









J. Burkhardt


about existing proposed legislation. I will, however, offer three modest
suggestions related to the concerns that have been raised regarding profes-
sional ethics, institutional missions, and our responsibility to posterity. (1)
Continued public funding for other kinds of research, including social sci-
ence and more holistic methodologies, may help address impacts before
they occur. (2) Continued risk-assessment utilizing the best results, not
only from environmental and ecological sciences, but also from the social
sciences, may give the broader perspective necessary for wise decisions
regarding biotechnology. (3) In a different vein, biotechnological review
committees at either national or university levels which include an ethics
and values component may both protect against shoddy research, as well
as provide greater legitimacy for well-conceived, soundly carried out bio-
technological research and development.
In summation, then, scientists and administrators I talked with believed
that the new biotechnologies are having, and will have, great social bene-
fits. They believe that the new biotechnologies may also carry great social
risks. I agree with both points. Concerns have been expressed about (1) the
impacts on professional values; (2) the mission of public research; (3) the
environment; (4) future science; and (5) effects on future generations. Fur-
ther research on these impacts is necessary, as is continued and improved
environmental risk-assessment. Education of both professionals and the
public as to the prospects and promises of biotechnology is also necessary.
Awareness about values and ethical conflicts brought about by the biotech-
nology revolution is also necessary. Future legislation should support the
work of social sciences and humanists in regard to this area of concern.
Where possible, future legislation should use, as a guide in decision-mak-
ing, the results of such research. In doing so, we may be able to guarantee,
as far as possible, that the broadest range of human and environmental
values will be considered and respected.










Testimony from Congressman Buddy MacKay

I would like to first thank Dr. Kozmetsky and the other persons who are
sponsoring this conference, and I would like to thank Dr. Glass. I wish my
colleagues could have heard that presentation. I think it would be certain
that the Congress, when it acts, would act in a knowledgeable way. My
comments will not be as broad as Dr. Glass', although I have to say that
even when you talk about a "narrow" part of this expanding issue, you are
talking about incredibly broad matters. President Lincoln, in the begin-
ning of a very famous speech, said, "If we could first determine where we
are and whether we are attending, we could better decide what to do and
how to do it." From my standpoint that threshold question is one that has
been inadequately addressed and I would like to spend some time talking
about that.
The spring issue of Foreign Affairs magazine has a major article by the
economist, Peter Drucker, in which he poses the thesis that we are in a
changed world economy. I would like to read three quotes just from the
first page. They set forth the thesis of my remarks today. He says,

The talk today is of a changing world economy. I
wish to argue that the world economy is not changing.
It has already changed its foundations and its structure
and in all probability the change is irreversible. It may
be a long time before economic theorists accept that
there have been fundamental changes and longer still
before they adapt their theories to account for them.
Above all, they will surely be most reluctant to accept
that it is the world economy in control rather than the
microeconomics of the nation's state on which most eco-
nomic theory still exclusively focuses. Practitioners,
whether in government or in business, cannot wait un-
til there is a new theory. They have to act and their
actions will be more likely to succeed the more they are
based on the new realities of the change in the world
economy.

That provides emphasis for the thesis of my remarks. The emerging issue
for the rest of this century is the need to compete in the world-wide mar-
ketplace. Dr. Glass touched on that in his overview. The new economy is a
world economy. It is an economy characterized by a quickening pace of
change, ceaseless innovation, and unprecedented flows of capital. The en-









30 B. MacKay
ergy source fueling the new economy is not oil or coal, but knowledge and
ingenuity. Raw material is not steel, but human brainpower. I will come
back to the energy source in a minute.
Against that background, it is clear that intellectual property rights as
an issue has changed from a dry, esoteric issue to a central issue, one of the
key issues determining whether America and American science will be
competitive in the world marketplace.
Coming back to the energy source, once again quoting President Lin-
coln, he made this statement that I think is remarkable, "The patent sys-
tem adds the fuel of self-interest to the fire of genius."
We are accustomed to that idea and America has a good system for
protecting intellectual property rights. It is somewhat in turmoil today as
Dr. Glass pointed out, and I think he pointed out in a thumbnail manner
the risks that are faced in our system by innovators, inventors, and small
firms particularly. A small firm has got to set aside a portion of its budget
for litigation and this system as it seeks to catch up with this wholly new
idea that life forms and living organisms can be patented and that people
can have property rights in that, part of it is going to be worked out in the
legislative branch in the Congress, part of it is going to be worked out in
the courts. Be that as it may, and given the turmoil and accepting as an
excellent thumbnail sketch what Dr. Glass has said about the places where
the questions remain to be resolved, internationally there is not even agree-
ment that there should be property rights and ideas. There is not even
agreement internationally that there are intellectual property rights much
less property rights and life forms and living organisms.
Some countries believe, and have the tradition, that ideas are the com-
mon property of mankind. The internationally recognized standard is that
foreign intellectual property rights should be treated the same as domestic
intellectual property rights, so some Third World nations have systemati-
cally weakened the laws protecting domestic international property rights.
They think weak laws will help them, so American firms and inventors
find themselves confronting an almost hopeless situation internationally-
inadequate laws defining the rights to be protected, inadequate lengths of
term for patents, some governments requiring licensing as a condition to
patent protection, inadequate anti-counterfeiting laws, and the absence of
the consensus in the international community on the desirability of en-
forcement. Clearly, things are not going to improve until the Third World
is convinced of the need to protect intellectual property rights and is will-
ing to enforce protection. Why is that important? It is a key element in the
emerging issue of competitiveness in America. Seventy-five percent of
American jobs are now vulnerable to foreign competition. If we are going









EXCERPTS FROM THE UNITED STATES SENATE HEARING


to avoid mounting domestic pressures for protectionism, government in
America is going to have to move aggressively to assure that America can
compete in the emerging world economy.
So the question is, what can the United States, which has the most at
stake, do to strengthen international protection of intellectual property
rights. Conceptually, we must make competitiveness a key element in
American foreign policy. What does that mean? It could mean these things
as opening ideas.

Withhold most favored nation status for any country that refuses to
enact and enforce strong.laws regarding intellectual property rights.
Re-think the negotiating strategy for bilateral agreements.
Give Third World nations a stake in intellectual property rights by
providing technical and financial aid if they will pass strong laws
protecting intellectual property rights.
Resist compulsory licensing as a condition to patent protection.
Expand the GAP agreement to prohibit counterfeiting and also to
cover intellectual property rights.

These are only beginning ideas. What else can we do? One of the things
that is happening in the House is the holding of joint hearings between the
Foreign Affairs Committee and the Science and Technology Committee in
which we are trying to look at this functionally for the first time in the
Congress. It may be that as the issue emerges, the Congress is going to have
to re-structure itself just so we can speak functionally about the problem.
The focus of the hearings is science and technology in foreign policy.
When you think about the emerging issue of economic competitiveness in
the same light as we now think about military competitiveness, you can see
that we have tremendous muscle and that we can enforce an order on the
world if we will only ourselves begin to think functionally about what our
problem is.
What else can we do? I don't know the answer to that. I am not sure it
has been thought through. I believe this hearing this morning and the
symposium that will follow will provide us with an excellent starting
point, and it is my pleasure to be a part of this.











Testimony from Dr. Donald R. Price
Vice President for Research
University of Florida

Biotechnology has become a major topic of discussion for the past sev-
eral years among university research administrators. The issues surround-
ing this new research field are numerous and many are extremely impor-
tant. The debate among and between the universities and private industry
has been rather fierce at times, but nearly always healthy and construc-
tive. I am of the opinion that the differences and more controversial prob-
lems are diminishing. A standard practice, or norm, is emerging that bet-
ter defines the issues and agreement is thus more easily attained.
The issues I am referring to are: (1) ownership rights to intellectual
property generated from a research grant or contract from an industry to
university researchers; (2) publication of research results without undue
delays and restrictions; (3) ownership rights to data; (4) confidentiality
requirements between universities and industry; and (5) patent licensing
agreements to commercialize results of university research.
In no technological field have the issues been more controversial than in
biotechnology. This is probably to be expected given the interest and eco-
nomic potential of new discoveries. There are numerous new start-up
companies all competing for survival. Because they are new and small,
they need the research base and support available at the universities. One
or two timely discoveries can make a company successful. Therefore, the
new companies are quite aggressive about control of patents and licensing
arrangements.
Let me proceed with a more detailed illustration of one issue that is
often at the forefront of disputes between industry and universities. Many,
in fact most, corporate-level officers in a company hold to the position that
if the industry provides funding for a research project on a university cam-
pus, then any discoveries (patents) that result from the research should
belong to the sponsor company. The universities take the position that if a
faculty researcher makes such a discovery, it belongs to the university. The
universities have very large investments in research faculty in the form of
laboratory space and equipment, libraries, graduate students, technicians,
etc. A faculty member usually develops expertise in a field over a period of
several years and it would be unfair for a private industry to come along
and reap all the benefits of that investment with one small research grant
or contract.
The university should, for this reason and others, retain ownership and
control of patents. As a public institution, we are obligated to get the










EXCERPTS FROM THE UNITED STATES SENATE HEARING


research from the lab out to benefit the public, if we can. We have an
arrangement to share any income that might result from a patent directly
with the inventors. Our policy is to share 50 percent of the first $100,000,
40 percent of the next $100,000, and 30 percent of all income over
$200,000 directly with inventors.
Now, to be fair with the industry that sponsors the research, we provide
in the research contract for the industry to have the first option to an
exclusive license to any patent resulting from the sponsored research pro-
ject. Usually when a potential sponsor fully understands our practice and
the rationale behind it, we have had very few disputes.
I believe the cooperation between industry and universities is good and
it is necessary to bring about the greatest benefits to the public. I hope the
federal government will continue to encourage this cooperation by contin-
uing the tax credits for industry support of research on university cam-
puses. This is a good investment in the future and needed to keep us com-
petitive with foreign countries.
Universities and industries can team up to help maintain our edge in
science and technology among the strong foreign competition. Biotechnol-
ogy will likely play a major role in this worldwide arena in the years
ahead.








PART II
ADVANCES IN
BIOTECHNOLOGY:
BREAKTHROUGHS AND
BOTTLENECKS










An Overview of Biotechnology

Philip H. Abelson
American Association for the Advancement of Science
Washington, D.C.

In this presentation, some of the progress in biotechnology will be evalu-
ated. The use of DNA technology in producing medical therapeutic and
diagnostic agents and the applications of monoclonal antibodies and tissue
culture will be discussed. The agricultural importance of rhizobia and
mycorrhyzae will be mentioned briefly.


DNA TECHNOLOGY

Therapeutic agents derived from the application of recombinant DNA
techniques have received much attention. Eventually, they will undoubt-
edly play an important role in medicine. Today, however, only three have
been approved by the U.S. Food and Drug Administration (FDA) for ther-
apeutic use. They are human insulin, growth hormone, and alpha inter-
feron. The histories of these pharmaceuticals provide perspective on time
tables and problems in creating proteins for therapy.


INSULIN

For more than 50 years, insulin extracted from the pancreases of cattle
or swine has been used in treatment of human diabetes. However, animal-
derived insulin is not identical to the human product, and in some in-
stances this may lead to unwanted reactions. In addition, supplies of beef
and pork insulin are limited, and a future scarcity has been envisioned.
Insulin is a relatively small and simple protein whose structure is well
known. Isolation of the gene giving rise to it was comparatively easy, and
human insulin was one of the first proteins to be produced by recombinant
DNA procedures. Escherichia coli were used in the process; the initial
creation of the bacterial product occurred in 1979. Insulin with a proper
human composition was an attractive product, with every reason for
speedy production, clinical testing, and FDA approval. Nevertheless, it
took seven years for human insulin, named Humulin, to achieve a signifi-
cant role in the management of diabetes.









P. Abelson


The full story of what happened has not been detailed. However, ru-
mors are that problems were encountered in the use of E. coli. The insulin
gene was well expressed, but the protein formed was retained within the
bacterial cells. Complete separation of the product from the host of other
E. coli proteins was absolutely essential to avoid pyrogenic activity, but
this was not easy. The initial sales price of Humulin was much greater than
that of animal insulin. Apparently, the production problems have been
overcome, possibly by using another organism such as yeast as an expres-
sion system. In any event, Humulin is now broadly accepted. At least half
of new diabetic patients in the United States are being treated with it.
Humulin now sells for less than animal insulin.
In summary, experience with insulin demonstrates that important thera-
peutic agents can be produced by recombinant DNA technology. Costs can
be competitive. The time required to go from laboratory to widespread use
can be many years.


GROWTH HORMONE

The second therapeutic recombinant DNA drug to be approved by the
FDA, a growth hormone for humans, also has a long history and favorable
circumstances for its acceptance. Genentech first produced its version of a
human growth hormone in 1979. Clinical trials began in 1981. At that
time, the human growth hormone being used for therapy was derived by
extraction of the pituitaries of human cadavers. The extracts were pooled
before final processing. Ultimately, concern was expressed that the prod-
uct contained harmful viruses, and in early 1985, the FDA cancelled ap-
proval of the human-derived growth hormone. This obviously had an ex-
pediting effect on the approval of the Genentech product which was
granted in October 1985. Some 4,000 children who would otherwise be
destined to be adult dwarfs are now being treated. It is estimated that
ultimately a total of the order of 10,000 to 15,000 children will receive the
drug. Costs are high. They range between $4,000 and $6,000 per patient
per year.
Patients treated with the Genentech drug have responded well. How-
ever, the protein is not identical to the human hormone. Other small com-
panies have created the human variety by recombinant DNA technology,
and their products are now being tested clinically. Genentech has found it
prudent to create its own new comparable product and is subjecting it to
clinical trials. Clinical trials are expensive. Costs quoted range from $25
million to $60 million.









AN OVERVIEW OF BIOTECHNOLOGY


Following clinical trials, applications for approval must be submitted to
the FDA. If some of the companies are not successful, they will suffer a
crippling financial loss. If all the companies are successful, the market will
be fragmented and probably competitive low prices will ensue. In that
case, nobody wins. The possible out is a non-therapeutic and unethical use
of the hormone in normal people in an attempt to increase their statural
growth.


THERAPEUTIC AGENTS FOR CANCER

The third recombinant DNA therapeutic drug approved by the FDA is
alpha interferon. The 1985 Biogen annual report stated that a Biogen-
produced alpha interferon was licensed in Ireland. On June 4, 1986, the
FDA announced approval of alpha interferon for use in treatment of hairy
cell leukemia. This is a rare form of cancer that affects 2,000 to 3,000
Americans. In contrast, about five million Americans have a history of
cancer. Treatment of the hairy cell leukemia with alpha interferon has
been quite successful. Remission of the disease has been noted in 75 to 90
percent of the more than 2,000 patients tested. But comparable effects
have not been seen in the treatment of major forms of cancer.
A substantial number of recombinant DNA therapeutic drugs are cur-
rently undergoing clinical trials. Besides alpha interferon, these include
beta and gamma interferons, interleukin-2, tissue necrotic factor, colony-
stimulating factors, tissue-type plasminogen activator, and a number of
others.
Cancer is such a dreaded disease that developments promising therapy
for it draw front-page attention. Recombinant DNA technology has re-
peatedly received great publicity and the consequent interest of financial
people. The first big wave of financial support for the emerging biotech-
nology companies was largely based on the thesis that interferons would
cure cancer. A large number of companies, including Genentech, Cetus,
and Biogen, cloned the genes and have produced interferons. A paucity of
news about success of the ongoing clinical trials indicates that the inter-
ferons have not lived up to the earlier high expectations. Rumor has it that
there have been serious side effects. One way of gauging what has hap-
pened, or is happening, is to examine annual reports of public companies
such as Biogen, Cetus, and Genentech. The reports of Cetus and Genen-
tech contain considerable detail. Genentech mentions studies in which a
combination of alpha and gamma interferons is employed. It also men-
tions the use of interferons combined with tumor necrosis factor. Another









P Abelson


clinical study involves interferons plus conventional chemotherapeutic
agents. The Cetus annual report for 1985 mentions clinical studies being
performed on beta interferon. It states that the product has been tolerated
well by patients and several responses were observed during the Phase I
testing, even though trials were designed to test safety and not efficacy.
The Cetus report put more emphasis on interleukin-2 as a cancer therapeu-
tic agent. It said that interleukin-2 is tolerated well by patients and that
early responses have been seen with Kaposi's sarcoma, chronic leukemia,
renal cell carcinoma, colon carcinoma, and others in Phase I studies. A
clinical study at the National Institutes of Health (Rosenberg, 1985) in-
volving 25 patients has testified to efficacy of interleukin-2. Cetus, how-
ever, is hedging its bets by developing a number of other therapeutic
agents for cancer, including tumor necrosis factor.
From the foregoing, it is evident that interferons alone are not magical
silver bullets for cancer. However, it is likely that recombinant DNA tech-
nology will produce additional useful therapeutic agents and that combi-
nations of them may prove fairly efficacious in some, but not all, forms of
cancer.


TISSUE-TYPE PLASMINOGEN ACTIVATOR

From the standpoint of successful treatment of patients, tissue-type plas-
minogen activator holds promise of becoming one of the most important
products of recombinant DNA technology. Heart attack is the leading
cause of death in the United States. An estimated 1.5 million Americans
will experience a heart attack this year, and 550,000 of them will die. To
function properly, heart muscle requires an adequate supply of blood.
When a clot forms in a coronary artery and blocks the flow of blood, a
heart attack may follow. If blood flow can be restored within an hour or
so, irreversible tissue damage can be stopped. Tissue-type plasminogen
activator is a potent natural clot-dissolving agent which is present in small
amounts in the circulatory system and acts to initiate normal clot-dissolv-
ing activities. However, when a massive clot forms, the body does not
provide a sufficient amount of the enzyme to dissolve the clot. Tissue-type
plasminogen activator is a protein of molecular weight 60,000. At least ten
companies have cloned the gene for it, and the product is undergoing
extensive clinical trials (Crawford, 1986). The product is proving superior
to an existing agent, a streptokinase. Prospects are said to be excellent for
early approval of the product by the FDA. Approvals will likely come one
at a time, with delays in between.









AN OVERVIEW OF BIOTECHNOLOGY


Some general remarks about recombinant DNA technology follow.
First, once a protein or its gene has been isolated, techniques are broadly
available for expressing the gene. The early dependence on E. coli has
vanished. Individual companies have available as many as ten expression
systems, including bacteria, yeast, molds, and mammalian tissue. A year
ago, a postdoctoral student told me, "Anybody can clone a gene." His
statement reflected the situation in some developed countries, but not in
most countries. An important bottleneck is availability of restriction en-
zymes and other substances including tagged biochemicals. Another state-
ment made to me was, "Genentech has cloned every gene that manage-
ment thinks might be of possible interest." That is not to say that
everything has been done, or even most of it. Rather, the statement is made
to indicate that as opportunities and needs arise, the necessary technology
will be quickly applied, provided a financial or other incentive is present.


PROTEIN ENGINEERING

For several years, the possibilities of changing the content of a gene have
been widely recognized. Progress has been made in this, some of which has
been reported in the literature (Villafranca et al., 1983). The procedure
thus far has been to modify a gene slightly so that when the gene is ex-
pressed, one of the amino acids has been replaced by another. Experiments
have shown that resultant enzyme activity may be enhanced, unchanged,
or decreased. Coincident with this, the folding of the protein and its shape
may be changed. Substantial progress is being achieved in understanding
the mechanisms, and top-notch crystallographers and extensive computer
calculations are involved. Already practical applications are emerging,
and many more can be expected. In its 1985 annual report, Cetus discloses
production of what it calls muteins of natural proteins. Cetus has found
that both the biological activity of the protein and the ease of production
can be enhanced by changing some of the amino acids. In particular, an
improved interleukin-2 was achieved by this procedure. Obviously, a new
chemical form is patentable, and this could be an additional crucial ad-
vantage. In the annual report of Genencor, genetic engineering of subtili-
sin is mentioned. This protein, which is a protease, has been engineered
and expressed in as many as 80 variants. An amino acid at one point in the
molecule has been replaced by all of the other 19 amino acids. Genencor
has been exploring the properties of subtilisin with a view to using it as a
protease in detergents. They have found that by replacing amino acids in









P Abelson


the natural enzyme, activity of the product can be enhanced. Other
changes noted include tolerance of pH, temperature, and oxidizing condi-
tions.
The combination of expertise permitting design of protein structures
with enhanced performance and the advantages of patents guarantee ex-
tensive and successful applications of genetic engineering to create new
non-natural proteins. When used in vitro, there should be only advan-
tages. However, when applied as therapeutic agents, some may have anti-
genic properties.


VACCINES

A major world public health problem is the lack of vaccines to immu-
nize large populations in the less developed countries. As a result, life
expectancies are 10 years and more below those of the developed countries,
and many people suffer debilitating diseases. In part, the lack of vaccines
results from inability to purchase them. In part, the problem is due to the
fact that needed vaccines have not been produced for important tropical
diseases.
Several U.S. companies have conducted research and development
aimed at producing hepatitis B vaccine. A recent announcement (Bialy,
1986) states that human trials of an antimalarial vaccine have begun at the
Walter Reed Army Hospital.
However, commercial biotechnology companies can be expected to give
a very low priority to producing vaccines under present-day circum-
stances. The financial incentives are, if anything, negative. Past experi-
ence is that virtually all vaccines can have serious side effects. Incidence of
these may be only one in 100,000 or less. But present-day damage suits
often result in awards of a million dollars or more. In consequence, many
makers of conventional vaccines have stopped producing them. To induce
competent companies in the United States to create and produce new vac-
cines will require mechanisms to eliminate threats of huge damage suits
and to provide financial incentives.
Under these circumstances, the developing countries should join cooper-
atively to create and produce vaccines to meet their needs. Following pro-
duction of these agents there would be practical problems to face, for
example, avoiding spoilage where refrigeration is non-existent and sanita-
tion is poor.
A recent development at the Wadsworth Center for Laboratories and
Research, New York State Department of Health in Albany, could be very









AN OVERVIEW OF BIOTECHNOLOGY


helpful (Perkus et al., 1985). Smallpox was eliminated through global use
of vaccinia virus. A dried preparation could be transported safely and
administered under primitive conditions. The New York workers have
found that other antigens can be expressed when vaccination is performed.
Using gene splicing, they incorporated additional antigens into the vacci-
nia virus. One successful formulation includes antigens of hepatitis B virus,
herpes simplex virus, and influenza virus hemagglutinin. The use of vacci-
nia virus is not completely without hazard. In one out of 300,000 in-
stances, very serious side effects occur. The New York group has entered
into collaboration with a French pharmaceutical company.
Prospects are good for development of many vaccines for veterinary
applications. Regulatory and legal problems may be encountered when
attenuated viruses are employed in vaccines. However, antigens produced
by recombinant DNA techniques should encounter less resistance. Several
years ago, Science awarded the Newcomb Cleveland prize for an out-
standing article on successful development of a protein-based vaccine for
one form of foot-and-mouth disease (Kleid et al., 1981). Annual reports of
the major biotechnical companies do not mention animal vaccines. How-
ever, a number of vaccines have been developed, and they are described by
Weppelman (this volume). In addition, at the University of Florida, work
is proceeding on the important tick-borne diseases anaplasmosis, babesio-
sis, and heartwater. Millions of cattle world-wide are at risk of these dis-
eases. The University of Florida is in a unique position to conduct research
on tropical diseases. It has an active center for tropical animal health,
expertise in biotechnology, and collaborative links with the Caribbean and
Africa.


DNA PROBES

An increasingly important application of DNA technology is its use in
diagnostic probes. These have been employed for some time, but recent
improvements in techniques have greatly increased their usefulness and
sensitivity. The technique applied in the past has been to lyse cells and to
absorb their DNA on a support (such as filter paper) in single-stranded
form. Subsequently, the strands are exposed to radioactively-tagged DNA.
If the two DNAs match, the radioactivity is retained. Otherwise, it is
washed away. Such a technique can be used to detect the presence of DNA
from any source, including viruses. The DNA-probe techniques are likely
to prove of special value in detecting viral plant diseases.









P Abelson


Cetus has recently obtained patents on a variant of the DNA probe that
appears to be highly sensitive and does not require the use of radioactive
isotopes. It thus is applicable in physicians' offices and other places where
radioactive isotopes are not available. Instead of radioactivity, biotin is
incorporated in the DNA probe. Biotin changes color when it interacts
with streptavidin. Cetus has also issued a press release mentioning a DNA
amplification procedure which they say achieves million-fold amplifica-
tion in target DNA. In consequence of these various developments, Cetus
has said that it has an enormously sensitive procedure for detecting DNA.


MONOCLONAL ANTIBODIES

Monoclonal antibodies already have been widely applied as diagnostic
aids and are beginning to have therapeutic applications.


DIAGNOSTIC AIDS

From the standpoint of improvement in the practice of medicine, mono-
clonal antibodies used as diagnostic aids have thus far had much more
impact than has recombinant DNA. As of May, 1986, the FDA had ap-
proved 140 diagnostic kits. These kits are used for in vitro tests, and hence
applicants need only demonstrate efficacy. The number of approvals has
been growing exponentially. In early 1983, the total number was about 20.
In late 1984, the number was 55. In part, the large figure results from
development by a number of companies of tests for the same organism, for
example, Chlamydia. Monoclonal antibodies are easily produced. Once
the relevant hybridoma is created, it can be immortal. Samples of it in-
jected into abdominal cavities of mice result in the production of fluids
containing antibodies for as many as 10,000 tests from each mouse. In the
future, the bottleneck will be marketing rather than production.
The two outstanding pioneers in commercialization of monoclonal anti-
bodies were Hybritech and Genetic Systems. These two started out as
small biotechnology companies. They succeeded in rapid development of
diagnostic aids. A pregnancy test by Hybritech was rapidly accepted. The
test detected pregnancy 10 days after conception. Genetic Systems (No-
winski et al., 1983) earlier produced diagnostic tests for venereal disease
that are quick and very useful. The two companies have developed many
other tests. A measure of their success and of the value of their products is









AN OVERVIEW OF BIOTECHNOLOGY


that both companies have become subsidiaries of major pharmaceutical
firms. In each case, the consideration involved was about $300 million.


THERAPEUTIC POTENTIAL

Those engaged in monoclonal antibody research are looking to frontiers
beyond diagnostic aids. Recently this has resulted in the FDA approval of
Orthoclone OKT*3, produced by the Ortho Pharmaceutical Corporation.
The drug has proved useful in treatment of short-term episodes of kidney
transplant rejection. Research teams are also developing therapeutic anti-
bodies for intractable bacterial infections and are seeking agents for diag-
nosis and treatment of cancers.
Some 60,000 patients in hospitals die each year as a result of opportunis-
tic infections by organisms that do not respond well to antibiotics. These
organisms, Klebsiella, Pseudomonas, and E. coli, produce potent endotox-
ins. These toxins can be neutralized by antibodies. However, mice cannot
be used to produce them. To avoid antigenic effects, the therapeutic anti-
bodies must be produced from human cell cultures. This is being done.
Monoclonal antibodies have been heralded as potentially important in
the diagnosis and treatment of cancers. Tumor cells produce antigens on
their surfaces. These can be used to create corresponding antibodies.
When such antibodies are injected into animals or humans, they tend to
bind to the tumor cells. This may interfere with the cells' metabolism.
Clinical trials are in progress testing this approach. Another procedure is
to tag the antibody with a radioactive isotope of yttrium. This combina-
tion can be employed to detect metastases of a cancer. Hopes have also
been raised about using the radiation from the isotope as a treatment mo-
dality. Another approach that has been tried is to couple a toxin such as
ricin to the antibody in the hope that the toxin will destroy the tumor cell.
These hopes have been around for several years, but conspicuous success
has not been evident. Apparently, the agent which includes ricin and anti-
body is not sufficiently specific in its binding, and it affects normal cells as
well as tumors. A procedure is now being investigated by Cetus in which
only the toxic part of the ricin is attached to the antibody. This approach
may prove more effective and less toxic to normal cells.









P. Abelson


PLANT BIOTECHNOLOGY

Technology has been applied to agriculture and the preparation and
preservation of food for a long time. Great improvements in yield and
quality have occurred. In many respects, in the short-term, the new tech-
nologies will enhance rather than supplant the old. For example, conven-
tional methods of plant selection, breeding, and field testing will not be
abandoned. In comparison to our knowledge about human physiology and
biochemistry, we are ignorant about the plant world. We are especially
lacking in knowledge about the role of the microbial life that exists in the
soil surrounding the roots of plants. We talk about altering the biochemis-
try of plants, but until recently have known little of the complex interac-
tions that control how genes are turned off or turned on.


TISSUE CULTURE

The big recent development in plant biology is tissue culture. It is per-
mitting the cloning of selected and virus-free plants. It permits more rapid
improvement of many plants than could otherwise be achieved. Originally
only a few dicotyledonous plants could be propagated in tissue culture, but
the number is increasing rapidly. Culture of monocotyledonous plants
from single cells has not proceeded well, but Winston Brill of Agracetus
believes that eventually all plants will be propagated through tissue cul-
ture. Such an achievement would open the road to large-scale enhance-
ment of the capabilities of plants.
Tissue culture is a necessary step in manipulations designed to select or
achieve an improved plant. By exposing tissue cultures to adverse condi-
tions such as salinity or pesticides, one can select those specimens most
capable of meeting the adverse condition.


GENETIC ENGINEERING OF PLANTS

Tissue culture also is involved in the introduction of DNA into cells to
change their genetic inheritance. Calgene has succeeded in introducing a
gene that gives tobacco some resistance to the herbicide glyphosate. Gly-
phosate, also known under the trade name "Round-up," is a successful
product of Monsanto. The chemical blocks a pathway in plants in the
synthesis of aromatic amino acids. The gene that Calgene has introduced









AN OVERVIEW OF BIOTECHNOLOGY


enables the plant containing it to continue synthesis of the aromatic acid in
the presence of Round-up.
In the experiments described to date, the resistance to Round-up is
somewhat limited (Marx, 1985). An additional enhancement of resistance
would be necessary to achieve an excellent plant that could thrive in the
presence of heavy applications of Round-up.
, A group at Monsanto has engineered glyphosate-resistant petunia plants
by inducing them to make 20 to 40 times the EPSP enzyme that is the
target of glyphosate (Marx, 1985). This was done by attaching petunia
EPSP synthase gene to a viral-regulating sequence that is a promoter of
gene expression and then transferring the hybrid gene into the plants.
Recently, I learned of another instance of a successful introduction of a
gene into a plant. My informant was Professor Lawrence Bogorad of Har-
vard University. He often collaborates with Belgian investigators and was
in Belgium recently to see results of a trial of incorporation of a toxin gene
into tobacco. The gene was derived from Bacillus thuringiensis, and a
toxin elaborated by this gene is known to be highly toxic to caterpillars
while not affecting other forms of life. The Belgian scientists had not only
incorporated the gene into the genome of the plant, but they had managed
to incorporate it in such a way that the gene was highly expressed. In
perhaps an overstatement, but a colorful one, Bogorad said, "The caterpil-
lars took one bite out of the plant and fell over dead. The control plants
nearby were reduced to shreds."
The superior expression of the B. thuringiensis gene in tobacco contrasts
with the situation that usually occurs in humans. In all our cells we have
the same DNA, yet only a fraction of the genes are turned on, and those
that are turned on change during development. In one case of the early
work on recombinant DNA, a great amount of information was available
about E. coli. It was possible to use that information to achieve enormous
expression of a foreign gene by including it in a plasmid that carried func-
tions essential to the growth of E. coli. In experiments with protoplasts,
only a small fraction incorporate DNA from the medium into their genome
and that DNA may or may not be expressed. However, investigators are
learning how better to incorporate genes and to get them better expressed.
In any event, the simple culture of plant cells is having and will have
major world-wide benefits. The technique is simple, the chemicals needed
are readily available, and scientists in the developing countries can obtain
and use them. Each terrain in each country presents a different circum-
stance, and the varieties of plants that can be selected and grown by tissue
culture are innumerable. Active programs in tissue culture are being car-









P. Abelson


ried out in many developing countries, including Brazil, Colombia, Costa
Rica, Cuba, Mexico, China, India, and Thailand.


RHIZOBIA AND MYCORRHYZAE

Plant biologists have long been aware of the role of rhizobia in nitrogen
fixation and of beneficial effects of mycorrhyzae in capturing phosphate.
Recent work has led to more systematic and effective use of these microor-
ganisms. The results are particularly applicable to developing countries
that are chronically short of hard currencies to pay for fertilizers. Rhizobia
naturally present in a soil are not necessarily optimum for the legume that
has been planted. On a recent visit to Brazil, I was told of a problem
encountered when farmers attempted to grow soybeans in a region of the
Cerrado. Nitrogen fixation was poor. The problem was traced to an Acti-
nomyces that secreted an antibiotic injurious to the rhizobia. Through
selection, a variety of rhizobia was obtained that is resistant to the antibi-
otic, and yields of soybeans improved. Considerable research is being con-
ducted in other developing countries to discover improved varieties of rhi-
zobia. Examples are India and Kenya. In Kenya, the effort has proceeded
to the point where 10,000 farmers have been provided with rhizobium
inoculants.
Mycorrhyzae associated with plants can have very important beneficial
roles. Benefits can include enhanced efficiency of uptake of phosphate,
drought tolerance, broader pH tolerance, and resistance to certain patho-
gens. These fungi attach to plant roots and send out hyphae that increase
by, for example, a factor of ten the volume of soil tapped by the plant.
Many woody plants have ectomycorrhyzal fungi as their symbionts. Tech-
niques are available to grow the mycelia of these fungi in culture, and they
are commercially available in the United States. Millions of pine seedlings
are already being treated. Benefits include enhanced performance of seed-
lings in the nursery and better survival and growth in the field.
The symbionts of most food crops and some trees invade the roots, re-
sulting in combinations called vesicular-arbuscular mycorrhyzae (VAM).
Until just recently, inocula were not commercially available. However,
Native Plants, Inc.(NPI) of Salt Lake City, Utah, has announced that it
can provide inocula. The NPI product is in the form of spores. Recently I
visited the laboratory of a subsidiary of NPI called Bio-Planta located in
Campinas, Brazil. There I saw a demonstration of the efficacy of VAM in
promoting the growth of lemon tree seedlings. The controls were only
about half as tall as the treated specimens, and the controls did not look










AN OVERVIEW OF BIOTECHNOLOGY


healthy. The scientist who showed me the seedlings said that he had tested
80 different isolates of VAM. Of these, six performed very well. The re-
mainder were not much better than controls. Evidently much is to be
gained in the health of plants and their growth by application of rhizobia
and mycorrhyzae. In addition, the optimum symbiont for a given crop is
probably a function of local conditions. Ultimately, it may be possible to
engineer better symbionts, but it is also desirable to understand better the
treasures that nature has already provided.


PERSPECTIVE AND CONCLUSIONS

Much of the future of biotechnology will be shaped by legal and regula-
tory considerations. In turn, the regulatory environment will be shaped by
the tides of public opinion. It is probable that when there is a clear-cut
obvious benefit, such as cure for a disease, the public will applaud. How-
ever, the public is easily made apprehensive, and proposals to release engi-
neered microorganisms into the environment are likely to encounter tough
going.
Probably the biggest factor shaping the future of biotechnology will be
the patent situation. An enormous number of patents have been applied
for and doing so has become highly fashionable. The 1984 report of
Genentech stated that they had received 100 patents and had 2,000 appli-
cations pending. The 1985 report of Cetus mentioned 1,000 applications.
Cetus press releases indicate that they have been successful in quite a few
applications. Annual reports of other companies mention patents. It is
likely that in coming years a considerable troop of lawyers will dine well
on biotechnology.
Tremendous progress is being made in the application of fundamental
knowledge. But because of the need for caution when preparing and ad-
ministering therapeutics, the time required will be long for full demonstra-
tion of the power of recombinant DNA technology. Gene engineering is
here, and it holds great promise. Monoclonal antibodies are already im-
proving the practice of medicine. Tissue culture has been sufficiently suc-
cessful that there is no doubt that it will have a great impact on world
agriculture. The study and use of soil microorganisms will likewise make a
major contribution. Those engaged in biotechnology are active in an area
where great things have happened and will continue to happen.









P. Abelson


REFERENCES

Bialy, H. (1986). Cloned malaria vaccine enters the clinic. Biotechnology
4: 384.
Crawford, M. (1986). Biotech market changing rapidly. Science 231: 12-
14.
Kleid, D.G., Yansura, D., Small, B., Dowbenko, D., Moore, D.M.,
Grubman, M.J., McKercher, P.D., Morgan, D.O., Robertson,
B.H., and Bachrach, H.L. (1981). Cloned viral protein vaccine for
foot-and-mouth disease: Responses in cattle and swine. Science
214:1125-1129.
Marx, J.L. (1985). Plant gene transfer becomes a fertile field. Science 230:
1148-1150.
Nowinski, R.C., Tam, M.R., Goldstein, L.S., Kuo, C.C., Corey, L.,
Stamm, W.E., Handsfield, H.H., Knapp, J.S., and Holmes, K.K.
(1983). Monoclonal antibodies for diagnosis of infectious diseases in
humans. Science 219: 637-644.
Perkus, M.E., Piccini, A., Lipinskas, B.R., and Paoletti, E. (1985). Re-
combinant virus: Immunization against multiple pathogens. Sci-
ence 229: 981-984.
Rosenberg, S. (1985). Observations on the systemic administration of the
autologous lymphokine-activated killer cells and recombinant in-
terleukin-2 to patients with metastatic cancer. New England J.
Med. 313: 1485-1492.
Villafranca, J.E., Howell, E.E., Voet, D.H., Strobel, M.S., Ogden, R.C.,
Abelson, J.N., and Kraut, J. (1983). Directed mutagenesis of dihy-
drofolate reductase. Science 222: 782-788.










Impacts of Biotechnology on Agriculture:
Plants

Peter R. Day
Plant Breeding Institute
Cambridge, England


BACKGROUND

This paper is concerned with plant molecular biology and agricultural
crop plants. Although the first topic is too young to have had a major
impact on the second, it is such a vigorous and rapidly-developing science
that many people expect that this will occur soon. Depending on the nat-
ure of their interests, they want to be sure not to miss new opportunities to
benefit from applications, and are anxious to monitor progress in the field
and even influence the directions this may take. Even the most sober en-
thusiasts, and I count myself among them, recognize that the impact of
molecular biology on our understanding of genetics and development
could well have profound implications for those whose job it is to shape
new materials for food and fibre. The early pipe dreams of nitrogen-fixing
cereals, and entirely new ranges of crops and farm animals, have been put
on the shelf for the time being, as many practicing scientists grapple with
the realities.
During the years since the Second World War, the development of agro-
nomic crops has made tremendous strides. In the developed world nearly
all of the major crops, except perhaps for soybean, have shown a steady
increase in yield which, in the last few years, has helped to generate, in
North America and the European Community, embarrassing surpluses.
There is no shortage of food in the developed world, and the difficulties of
poorer nations are often seen as primarily due to the problems of transport-
ing grain to where it is needed and of paying for it. Speaking as a plant
breeder, there are two major challenges facing us at the end of the eighties.
These are, to provide the crop varieties needed to supply food and raw
materials for industry in the developed world, and to assist the less devel-
oped nations to become self-sufficient in agriculture by increasing produc-
tivity without increasing their dependence on chemical inputs. No matter
which part of the world he works in, the breeder has to attend to three
requirements: to increase yield, to improve the quality of the harvested









52 P. Day
crop, and to reduce production costs. This last includes resistance to pests
and diseases, and environmental stress, relieving the need for applied pes-
ticides and increasing the productivity of land areas and climatic zones not
suited to growth and development of presently available crop varieties.
Breeding crop plants is a very successful operation that relies on good
organization and management to handle large numbers of plants, sus-
tained hard work, and the exercise of judgement born of experience. Suc-
cessful breeders will use every trick and method they can find if they will
help them to carry out their tasks more efficiently (Day, 1985). They are
generally too busy to write many research reports or to spend time theoriz-
ing. If biotechnology can help them, they will be the first to want it, but
they are understandably skeptical of the claims made by many of its expo-
nents. It is important to be aware of this gap. It stems from the nature of
the day-to-day work and interests of breeders and molecular biologists.
The two disciplines use different terms. They involve work at either end of
the spectrum extending from very large plant populations to cells, organ-
elles, and molecules. A constructive dialogue does not necessarily occur
automatically. Biotechnology provides a method of painstakingly disas-
sembling the individual component parts of an organism and putting them
together again. Anyone who has ever assembled a radio or similar electri-
cal circuit with the expectation that it will work will know that it is neces-
sary to have a circuit diagram, showing the components needed, how they
should be laid out, and how they are wired together. Although the analogy
quickly breaks down, since a living organism is an assembly of very com-
plicated self-replicating circuits that have taken millions of years to evolve,
plant molecular biology is still very much at the stage of elementary tinker-
ing. This consists of taking things apart to see if and how they work when
put back together and inserted, more or less at random, into a functioning
organism such as a bacterium or yeast. All molecular biologists recognize
that a very great deal more has to be done to improve our understanding of
how plants work before we can be in a position to introduce the major
changes that will no doubt be possible sometime in the future.


CURRENT ACHIEVEMENTS IN PLANT
MOLECULAR BIOLOGY

This is neither the time nor the place for a comprehensive review of
progress; however, we note that for crop plants like tobacco, potato, to-
mato, sunflower and oilseed rape (canola), the introduction of a foreign
gene by transformation, using vectors based on the crowngall bacterium,









IMPACTS OF BIOTECHNOLOGY ON AGRICULTURE: PLANTS 53

Agrobacterium tumefaciens, is now almost commonplace. All of these di-
cotyledons are natural hosts for Agrobacterium. The frequency of trans-
formation, and the stability of transformants in subsequent generations,
are both sufficiently high to be potentially useful to plant breeders. But
important problems remain. As I shall mention later, there is no control at
present over the sites of integration of introduced DNA or of the numbers
of copies which become integrated. This means that each transformed
plant is potentially unique in regards to its fitness and field performance.
For cereals and many other monocotyledons there are still major diffi-
culties in effecting transformation. Until this can be done routinely, we
will not be able to exploit the benefits of gene transfer in wheat, barley,
maize, rice, and sorghum. Although it is possible to introduce DNA to
protoplasts, using the method of electroporation, regeneration of plants
from cereal protoplasts is proving to be extremely difficult technically.
Microinjection of fertilized wheat ovules appears to be a promising
method, since the tissue is programmed to develop into a seedling, but
work to date has been unsuccessful. Lorz (personal communication) re-
cently claimed to have introduced a bacterial gene for antibiotic resistance
into rye by injecting a DNA preparation into shoot meristems. However,
this, like earlier claims of Soyfer et al. (1976) for transformation of barley,
needs confirmation.
Sidestepping the issue of cereal transformation, we may ask what valu-
able single genes are available for introduction. If the gene product can be
identified, there are several methods for recovering cDNAs prepared from
the messenger RNA which specifies the gene product, which can then be
used to probe for the gene itself in a DNA library of the organism in a
bacterium. Another method, called transposon tagging, depends on Bar-
bara McClintock's discovery of jumping genes. Namely, that if you can
identify a mutation in a character that is of interest, that is, the result of
the gene controlling the character being inactivated by the insertion of a
small DNA sequence called a transposon, then it is possible, by detecting
the base sequence of the transposon DNA, to identify the gene by a series of
probing steps. As a result of using these and other methods, a steadily-
increasing number of plant genes have been identified and characterized.
Of particular interest in this list are genes involved in the determination of
self-incompatibility in Brassica oleracea (Nasrallah et al., 1985) and Nico-
tiana alata (Anderson et al., 1986). The structure of these genes should
help to unravel the mystery of plant cell recognition systems that not only
regulate pollen-tube growth in the style, but plant interactions with para-
sites.
Self-incompatibility is being used very effectively to prevent self-fertil-









54 P. Day
ization for the commercial production of hybrid seed of kale, another bras-
sica. The work of Nasrallah et al. (1985) offers the promise of probes to
detect the incompatibility genotypes of seedlings and the eventual possibil-
ity of directed changes by transformation in other important brassicas such
as oilseed rape, where the potential for hybrid seed production is very
much greater. Genes for herbicide resistance are likely to have early practi-
cal applications since they allow the plant breeder the opportunity to con-
fer resistance to useful herbicides to which his crop is sensitive. This in-
volves a role reversal, since until now the herbicide chemist has looked for
chemicals that exploit the differential sensitivities of the crop he wishes to
protect, and the weeds he wants to kill. There are also encouraging signs
that we are beginning to understand the mechanism whereby genes are
controlled during development, although there is still much to be found
out about the nature of the signals which turn the controls on and off. An
example of this is the promoter sequence controlling the element attached
to the storage protein of beans, Phaseolus vulgaris (Sengupta-Gopalan et
al., 1985). When this gene and its promoter were introduced into tobacco,
and transformed plants flowered and set seed, the introduced gene was
only expressed at the time of seed formation. Even though the protein it
formed was inappropriate for tobacco, the promoter attached to the gene
evidently responded to signals that mark the onset of seed development
which, it would seem, beans and tobacco have in common. There is con-
siderable interest in introducing genetic information from microorganisms
into higher plants. One example is the gene for a toxic polypeptide, the
delta endotoxin of Bacillus thuringiensis, which has now been introduced
into tobacco plants, conferring resistance to tobacco horn worm and other
caterpillars. An alternative use for the same gene has been its transfer to a
strain of Pseudomonas fluorescens isolated from corn roots (See Watrud et
al. in Halvorsen et al., 1985). Applied as a dressing to corn seed, the engi-
neered bacterium colonizes the plant root surface and, in greenhouse tests,
provides protection against the larvae of corn ear worm in the soil. Some
work with virus genes has shown also that if a cloned cDNA of the gene
specifying the coat protein of the tobacco mosaic virus particle is intro-
duced into tobacco, it may confer resistance either by delaying disease
development or by preventing virus infection by an as yet undetermined
mechanism (Abel et al., 1986). Many plant viruses have single-stranded
RNA genomes. There is interest in transforming plants with genes which
are transcribed to produce an antisense single-strand RNA complementary
to the viral genome. Here the idea is that the two complementary RNAs,
one of viral origin and the other now of plant origin, will interact to pre-









IMPACTS OF BIOTECHNOLOGY ON AGRICULTURE: PLANTS 55

vent transcription and multiplication of the viral genome (Palukaitis and
Zaitlin, 1984, see also Baulcombe, 1986).
One important use of plant molecular biology in plant breeding is the
production of specific probes, which can be used to detect moderately
large blocks of genetic information in the breeder's plants. One of the best-
known examples is the dot blot or sap spot test for detecting the presence of
viroids and viruses in plants. The reagent used is a cDNA prepared to part
of the RNA genome of the pathogen. This is then labelled in such a way
that when it hybridizes to complementary RNA, and thus becomes bound
to sap spots squeezed out and baked onto a cellulose nitrate membrane,
these can be revealed when the excess unbound reagent has been removed
by washing. The same method can be used to discriminate among differ-
ent cytoplasmic male sterile lines of corn, which have characteristic differ-
ences in their mitochondrial DNA (Flavell et al., 1983) to identify blocks of
terminal heterochromatin in rye lines used as parents for producing the
wheat/rye hybrid triticale and for detecting a chromosome arm from rye,
present in a number of European wheat varieties, which has a deleterious
effect on baking quality (Hutchinson et al., 1985).
An interesting new development with great promise makes use of the
fact that when DNA is extracted from a plant and treated with a restric-
tion enzyme, it is cut into small pieces of characteristic size. If the re-
stricted DNA is separated by electrophoresis on an agarose gel, and
stained, it is usually revealed as a smear composed of many thousands of
fragments of different length, which have migrated to different distances
in the gel. However, by using a labelled DNA probe, for example, pre-
pared by cloning a cDNA prepared from the plant's messenger RNA, or a
fragment of DNA present in low copy number, and treating the gel so that
only those fragments that bind to the probe are revealed (this is called a
Southern blot), a much simpler banding pattern emerges. When different
individuals within a population are compared, it is frequently possible to
show that there are significant differences in the banding patterns. This is
because the sizes of the individual fragments produced by the restriction
enzyme vary, perhaps as the result of the deletion or addition of bases.
These differences, known as restriction fragment length polymorphisms
(RFLPs), can be used as markers in order to identify those parts of chromo-
some arms from which they are derived. RFLPs detected in the DNA from
blood or semen samples provide a rigorous method of proving identity in
forensic science or of familial relationship (Jeffreys et al., 1985). For plant
breeding, RFLPs show promise of providing a convenient and ready-made
system of markers to develop linkage maps for any crop plant without the
need for extensive genetic studies such as those carried out in corn and









56 P. Day
tomato (Burr et al., 1983; Bernatzky and Tanksley, 1986). Thus, once char-
acters that are of interest to breeders have been associated with RFLP
markers, it should be possible to select only those progeny in segregating
families which have the maximal number of characters the breeder seeks.
This could reduce the number of backcrosses needed to introduce new
characters from an alien wild species and also reduce the number of gener-
ations needed in pedigree selection programmes. Since the markers are co-
dominant they may also be used to identify true breeding homozygotes. At
the present time the method is laborious. It will be much more widely used
when machines are developed to enable restriction digests, electrophoresis,
Southern blotting, and gel recording to be carried out automatically.
Once a gene has been isolated and cloned, it is of course possible to
modify its structure by altering the base sequence in places. By this means,
it is possible to direct changes in the amino acid sequence of a peptide,
with a view to changing the properties of the gene product. This may well
be useful in breeding programmes concerned with improving quality. An
example is breeding for improved baking quality in wheat, where wheat
breeders seek to maximize endosperm protein glutenin sub-units with ter-
minal sulphur-containing amino acids that, through disulphide-bonds,
contribute to the cross-linking that occurs between these proteins in the
development of dough required for bread-making (Day et al., 1986). Fig-
ure 1 summarizes the principal interactions that are likely between con-
ventional plant breeding and biotechnology.


CURRENT SCIENTIFIC ISSUES

Plant molecular biology is clearly a scientific discipline which has pro-
found long-term implications for plant breeding. However, I have alluded
to the difficulties in the way of achieving our expectations. One problem
that tends to be neglected is the need to establish a productive dialogue
between the individuals involved in the two activities. At the Plant Breed-
ing Institute, we believe this is best handled by developing the technology
that lies between them. At the moment this is largely concerned with the
plant tissue culture needed to raise newly-transformed plants for evalua-
tion and testing in collaboration with the plant breeders. The tissue cultur-
ist works with protoplasts, callus tissues, meristems, and embryos to select
and regenerate potentially useful transformed plants and must therefore
communicate effectively with the molecular biologists on whom he de-
pends for genetic constructs. He is thus aware of the breeder's require-









IMPACTS OF BIOTECHNOLOGY ONAGRICULTURE: PLANTS 57


1.


4. Test new lines
(breeding)


3.






9. Gene insertion
into tester stocks
(biotechnology, breeding)
T


8.


Current va
(agronom


Cross to adapted
varieties
(breeding)


5. Germplasm, landraces,
alien species (breeding,
genetics, conservation)

rieties
y)

2. Identify useful
characters
(breeding)
/ I


d



6. Identify genes
(genetics, physiology,
biotechnology)


7. Gene isolation
S(biotechnology)

Gene modification
in vitro
(biotechnology)


Figure 1. Breeding cycle showing the technologies used now (solid lines)
and that are likely to be used during the next decade (dotted lines). Modi-
fied from Austin (1986).


ments and time-scale, but is also familiar with those of the molecular biol-
ogist. This work of technology transfer will be crucially important for
success.
It is also clear that, for the foreseeable future, the application of bio-
technology to shape new forms of crop plants will continue to depend very
heavily on conventional plant breeding methods. Once new and interest-
ing material begins to flow in their direction, plant breeders will evaluate
it and incorporate some of it in their breeding programmes. There are


Q









58 P. Day

many reasons why they will be cautious. For example, at the present time
each product of transformation using the Agrobacterium system is poten-
tially different from every other. This is because as yet there appears to be
no control over the sites of integration of introduced DNA or the number
of copies introduced. In a plant breeding context, therefore, the effect of
integration site will have to be examined. Even if the molecular biologist
has appeared to 'gild the lily' by adding one or more new characters to an
established and widely-grown crop plant variety, the new line will need to
be tested for stability of the introduced characters) over several years and
many trial sites, to make quite sure that transformation has not introduced
an unanticipated defect which could render it valueless under certain cir-
cumstances. Plant breeders are used to uncovering the Achilles' heels in the
most cherished products of their selection programmes.
Perhaps one of the most important scientific and social issues concerns
framing sensible legislation to allow the introduction of products of genetic
engineering into agriculture outside the containment of the laboratory and
glasshouse, which is at present mandatory for all such materials (see
Halvorsen et al., 1985; Teich et al., 1985). The difficulties encountered in
arranging field tests of Ice-bacteria and of corn seeds treated with Pseudo-
monas fluorescens engineered to produce the delta endotoxin of Bacillus
thuringiensis kurstaki are well known. The companies interested in com-
mercializing these two microorganisms have spent millions of dollars, and
thousands of man-hours, attempting to proceed through a thickening tan-
gle of regulations and lawsuits. There can be no doubt that society must
insist on safeguards to protect the environment. However, such safeguards
have to be tempered by the knowledge that no-one can guarantee, with
complete certainty, that any given released organism, whether or not ge-
netically engineered, will have no adverse effects whatsoever. Answers
must be provided to certain questions before field tests and environmental
release are allowed. In my view, the requirement to produce, in an appli-
cation for an experimental use permit, more than a thousand pages of
evidence to show that an engineered organism is safe and harmless, would
be absurd if it is to become the norm. Unless procedures are simplified,
and sensible tests established, we will deny ourselves important benefits in
a technology that may well greatly reduce our dependence on agrochemi-
cals, which in the long run could be far more damaging both to us and the
environment than products of genetic engineering.









IMPACTS OF BIOTECHNOLOGY ONAGRICULTURE: PLANTS 59

SPECIFIC PRODUCTS AND TIME FRAMES

I may disappoint some readers by saying that I believe the time frames
for biotechnology, at least during its early years, will probably be little
different from those of conventional breeding. In our pedigree selection
breeding programme for winter wheat at the Plant Breeding Institute, it
takes from 10-12 years from the time a cross is made until the variety has
reached the Recommended List and is available in quantity for farmers to
buy and plant in their fields. By using the method of single-seed descent, it
is possible to speed up the production of homozygous lines from F3 to F6,
by cramming three generations into one year. However, since very little
selection can be practiced under these conditions, the method at best only
cuts two years off the total time. By using single-seed descent, our breeders
could expect to have identified a candidate variety for submission to Na-
tional List tests within five or six years. In Britain, National List tests take
two years, and are followed by a further year of testing for Recommended
List status. During this three-year period, seed stocks are built up to pro-
duce the 100 ton or so of basic seed needed for distribution to growers, who
will raise certified seed for sale to farmers. Let us suppose that, as a result
of genetic engineering, a plant breeder is presented with the equivalent of
his best current variety that is either already in commerce or is about to go
into the Recommended List, which now has a new engineered character,
perhaps either herbicide or disease resistance. The first question would be,
"Does the new form have sufficient merit to be a new variety in its own
right, with all or most of the attributes of the original variety, plus the
added character of resistance?" Even if the answer appears to be yes, the
breeder will need two years of trials to satisfy himself on that point, and
before the variety reaches a Recommended List, three more years of trials
will be necessary. By that time, the original variety will already have been
on the market for at least four or five years and may be outmoded by other
developments from conventional breeding. New varieties that the engi-
neered variety must compete with, might for example, have better quality
or resistance to other diseases that are now more important. The breeder
will have anticipated that this is likely to happen, and if he likes the mate-
rial from the molecular biologist, will have introduced it into his own
breeding programme to recombine the new character with the other fea-
tures he is working towards. However, the product of genetic engineering
may well take up to eight years to reach the farmer's field by following this
route. It follows from all of this that it might be more profitable in the
short-term to seek other ways of genetically engineering crop plants. For
example, why not grow your crop in the field and introduce some benign









P. Day


systemic virus, which carries a package of genes that have the effect of
promoting growth, altering the storage components in the endosperm of
the seed, and which confer resistance to a range of fungal, bacterial, and
insect pests. A step in this direction has already been taken by work to
develop virus vectors. Brisson et al., (1984) showed that a bacterial gene
for drug resistance, introduced into the genome of cauliflower mosaic vi-
rus,-was expressed in turnip plants systemically infected with the engi-
neered virus. I won't go into the question of which of these two kinds of
genetic engineering and applications would be the easiest to protect from
the point of view of patenting. I don't think either of them would be
particularly easy, although I suspect that engineered plants would more
easily satisfy regulatory authorities than engineered viruses. Other oppor-
tunities for genetic engineering may well involve the production of arable
crops which have entirely new properties from those we have at present.
Not many years ago, we were intrigued by the success of chemists reshap-
ing soybean protein into fibres that simulated the muscle fibres of meat.
Perhaps one day we will be able to engineer tubers from a potato-like
plant, which have a high content of both protein and fibre, to produce an
entirely new textured form of vegetable to replace the meat in our diet.
New developments of this kind may take longer but there will likely be no
alternative but to wait for them.


CONCLUSIONS

The problem with any discussion on the impact of plant molecular biol-
ogy on crop plant agriculture is that it soon degenerates into the realm of
science fiction. I hope I have indicated for the general reader my enthusi-
asm for, and high expectations of, the very exciting work now going on in
this field. Almost every week, in the journals that pour into libraries
around the world, there are new discoveries and new vistas opening up
which will give us cause to ask, "Is progress faster than we thought? Are
the barriers to what is possible more easily surmountable than we
thought?" I believe this will be the case, but I also believe that it would be
foolish to neglect, in the meantime, the tried and tested technology on
which we now depend for our food and fibre.


REFERENCES

Abel, P.P., Nelson, R.S., De, B., Hoffmann, N., Rogers, S.G., Fraley, R.T.









IMPACTS OF BIOTECHNOLOGY ONAGRICULTURE: PLANTS 61
and Beachy, R.N. (1986). Delay of disease development in
transgenic plants that express the tobacco mosaic virus coat protein
gene. Science 232: 738-743.
Anderson, M.A., Cornish, E.C., Mau, S.L., Williams, E.G., Hoggart, R.,
Atkinson, A., Bonig, E., Grego, B., Simpson, R., Roche, P.J., Ha-
ley, J. D., Penschow, J.D., Niall, H.D., Tregear, G.W., Coghlan,
J.P., Crawford, R.J. and Clarke, A.E. (1986). Cloning of cDNA for
a stylar glycoprotein associated with expression of self-incompati-
bility in Nicotiana alata. Nature 321: 38-44.
Austin, R.B. (1986). Molecular Biology and Crop Improvement. Cam-
bridge University Press, p. 114.
Burr, B., Evola, S.V. and Burr, F.A. (1983). The application of restriction
fragment length polymorphism to plant breeding. In Genetic Engi-
neering 5 (J. K. Setlow and A. Hollaender, eds), pp. 45-59. Plenum
Press, New York.
Baulcombe, D.C. (1986). The use of recombinant DNA techniques in the
production of virus resistant plants. In Biotechnology and Crop
Improvement and Protection (P.R. Day, ed.), pp. 13-19. British
Crop Protection Council, Thornton Heath.
Bernatzky, R. and Tanksley, S.D. (1986). Toward a saturated linkage map
in tomato based on isozymes and random cDNA sequences. Genet-
ics 112: 887-898.
Brisson, N., Paszkowski, J., Penswick, J.R., Gronenborn, B., Potrykus, I.
and Hohn, T. (1984). Expression of a bacterial gene in plants by
using a viral vector. Nature 310: 511-516.
Day, P.R. (1985). Crop improvement: breeding and genetic engineering.
In Technology in the 1990s: Agriculture and Food (K. Blaxter and
L. Fowden, eds.), pp. 47-54. Royal Society, London.
Day, P.R., Bingham, J., Payne, P.I. and Thompson, R.D. (1986). The way
ahead: wheat breeding for quality improvement. In Chemistry and
Physics of Baking (J.M.V. Blanshard, ed.), Royal Society of Chem-
istry, London (in press).
Flavell, R.B., Kemble, R.J., Gunn, R.E., Abbott, A. and Baulcombe, D.
(1983). Applications of molecular biology in plant breeding: the
detection of genetic variation and viral pathogens. In Better Crops
for Food, Ciba Foundation Symposium 97, pp. 198-209. Pitman,
London.
Halvorsen, H.O., Pramer, D. and Rogul, M. (1985). Engineered Orga-
nisms in the Environment: Scientific Issues. American Society for
Microbiology, Washington.
Hutchinson, J., Abbott, A., O'Dell, M. and Flavell, R.B. (1985). A rapid









62 P. Day
screening technique for the detection of repeated DNA sequences in
plant tissues. Theoret. Appl. Genet. 69: 329-333.
Jeffreys, A.J., Wilson, J. and Thein, S.L. (1985). Individual-specific 'fin-
gerprints' of human DNA. Nature 316: 76-79.
Nasrallah, J.B., Kao, T.H., Goldberg, M.L. and Nasrallah, M.E. (1985).
A cDNA clone encoding an S-locus specific glycoprotein from Bras-
sica oleracea. Nature 318: 263-267.
Palukaitis, P. and Zaitlin, M. (1984). A model to explain the 'cross-protec-
tion' phenomenon shown by plant viruses and viroids. In Plant
Microbe Interactions (T. Kosuge and E. Nester, eds.), pp. 420-429.
Macmillan, New York.
Sengupta-Gopalan, C., Reichert, N.A., Barker, R.F., Hall, T.C. and
Kemp, J.D. (1985). Developmentally regulated expression of the
bean b-phaseolin gene in tobacco seed. Proc. Nat. Acad. Sci. USA
82: 3320-3324.
Soyfer, V.N., Kartel, N.A., Chehalin, N.M., Titov, Y.B., Ceincinis, K.K.
and Turbin, N.V. (1976). Genetic modification of the waxy charac-
ter in barley after an injection of wild-type exogenous DNA. Analy-
sis of the second seed generation. Mutation Research 36: 303-310.
Teich, A.H., Levin, M.A. and Pace, J.H. (eds.) (1985). Biotechnology and
the Environment: Risk and Regulation p. 201. Amer. Assoc. Adv.
Science., Washington, D.C.










Impacts of Contemporary
Biotechnology on Animal Science

Roger M. Weppelman
Merck, Sharp & Dohme Research Laboratories
Rahway, New Jersey


INTRODUCTION

The term "Biotechnology" is not easily defined. By the more inclusive
definitions, animal science itself qualifies as one of the oldest (and most
successful) of the biotechnologies. Rather than attempting to define "Bio-
technology," I will simply state at the onset that the scope of this manu-
script will be restricted to those aspects of contemporary animal science
which have evolved from one or more of the following three papers in the
scientific literature:
1. Chung and Cohen (1974). This paper describes the use of a restriction
enzyme to create a functional genetic element bearing genes from
two species of bacteria and thereby marks the beginning of genetic
engineering.
2. Kohler and Milstein (1975). The authors fused a normal mouse spleen
cell secreting a single type of antibody with an immortal mouse
myeloma cell to produce the first hybridoma, which continued to
secrete antibody and was immortal. This was the start of hybridoma
technology and monoclonal antibodies.
3. Palmiter et al. (1982). These authors describe the creation of
transgenic mice. Strictly speaking, this was not the first creation of
transgenic mice, but because the gene used was beautifully engi-
neered, because the technique was elegant and widely applicable,
and because the results were truly dramatic, this paper marks the
beginning of transgenic animals.


GENETIC ENGINEERING

Restriction enzymes are the basis for genetic engineering because of
their unique ability to cleave double stranded DNA in such a way that the
pieces can be readily rejoined (Maniatis et al., 1982; Rodriquez and Tait,
1983). Hybrid genes can be engineered by cleaving two unrelated chromo-









64 R. Weppelman
somes with a restriction enzyme and then rejoining pieces of one chromo-
some with those of another. Insertion of a properly constructed hybrid
gene into a suitable organism will enable the organism to produce the
protein specified by the hybrid gene. If the protein is an enzyme, then the
organism's metabolism might be altered to produce new metabolic prod-
ucts, or perhaps old products in better yields. In a conceptual sense, ge-
netic engineering has the potential to yield the following classes of prod-
ucts: (1) recombinant genes; (2) organisms containing recombinant genes;
(3) proteins made from recombinant genes; and (4) the products of en-
zymes produced from recombinant genes.
The product class comprised of recombinant genes will probably find
application in correcting genetic defects in humans but is unlikely to be
widely used by the animal industry. The second product class, organisms
bearing recombinant genes, is discussed below under Vaccines and
Transgenic Animals. A third possible product of this class is rumen micro-
organisms engineered to improve the efficiency of fermentation. The final
class listed above, the products of enzymes specified by recombinant genes,
could be extremely important since many products currently marketed for
the animal industry are produced by microorganisms during fermentation.
Among these are the coccidiostats and rumen additives monensin, salino-
mycin and lasalocid, the anthelmentic ivermectin, and numerous antibac-
terials, vitamins and essential amino acids. Insertion of the appropriate
genes into the production microorganisms could lead to more efficient fer-
mentations or to the production of improved derivatives of the original
drug.
The type of potential product which has caused the most excitement to-
date is the class comprised of proteins synthesized from recombinant DNA.
Before genetic engineering, most of these proteins were available in
amounts which were adequate for only the smallest of laboratory experi-
ments. These proteins are now available in amounts sufficient for large-
scale studies of their activities and they have the potential to become avail-
able in the quantities needed to satisfy the needs of the world's animal
industry. Three general types of proteins produced by recombinant DNA
technology are discussed below.


HORMONES

The ability of bovine pituitary hormones to stimulate dairy cattle to
produce more milk was suggested in 1937 by the observation that extracts
prepared from bovine pituitaries enhanced milk yield (Asimov and Kouze,









IMPACTS OF CONTEMPORARYBIOTECHNOLOGY-ANIMAL SCIENCE 65

1937). This effect was intensively studied in Britain as a way of increasing
milk supply during World War II (Young, 1947). Subsequently, the active
component within the pituitary extracts was shown to be bovine growth
hormone (Brumby and Hancock, 1955). This observation remained only a
curiosity until the gene was cloned and the hormone was produced in
quantity by microorganisms. A recent publication by Bauman et al. (1985)
demonstrated that methionyl bovine growth hormone, made by E. coli,
increased milk production over a 188 day period by 23 to 41 percent,
depending on the daily dose. Just as importantly, the efficiency with which
feed was converted to milk improved up to a maximum of about 10 per-
cent. These data indicate that bovine growth hormone could enable the
dairy industry to satisfy the current demand for milk with an estimated 30
percent fewer cattle and at a considerably reduced cost in feed (Animal
Pharm, November 14, 1985, p. 15).
It should be noted that the increases in milk production induced by
growth hormone did not lead to net losses of body weight. Indeed the
cattle gained weight while being treated with growth hormone and the
gains were comparable to those of untreated cattle during the same period.
This observation should put to rest concerns that growth hormone will
strain dairy cattle by causing them to produce excessive milk (Biotechnol-
ogy News, April 18, 1986, p. 1).
A very novel alternative use of bovine growth hormone to stimulate milk
production was recently patented by Bauman and Sejrsen (U.S. Patent No.
4,521,409). These inventors discovered that treatment of dairy cattle dur-
ing adolescence stimulates mammary development and increases milk out-
put after their first calf. Whether treating with additional growth hor-
mone during lactation will further enhance milk production remains to be
determined (Genetic Technology News, July 1985, p.3).
The role of growth hormones in regulating growth has been recognized
for years and for most species reductions in circulating growth hormone
levels have been unambiguously associated with reduced growth. How-
ever, until very recently it was questionable whether giving additional
growth hormone to "normally" growing animals would improve either
their growth or utilization of feed. The answers for the important stock
animals have been mixed. Increasing growth hormone levels in chickens
three- to ten-fold did not affect growth rate (Souza et al., 1984). When
growth hormone purified from ovine pituitaries was given to growing
wether lambs, growth was improved slightly but not significantly and effi-
ciency was improved significantly by about five percent (Muir et al.,
1983). In a more recent experiment (Johnsson and Hart, 1985), treatment
of growing female lambs with growth hormone from bovine pituitaries









66 R. Weppelman

significantly stimulated both gain and efficiency by 22 and 12 percent,
respectively. These authors noted that the hormone caused a 50 percent
increase in fleece weight. Dramatic effects of growth hormones have also
been reported in swine. When Baile et al. (1983) treated growing pigs with
daily doses of recombinant human growth hormone, growth was stimu-
lated up to 10 percent, but efficiency was not affected. Chung et al. (1985)
reported that daily doses of growth hormone from porcine pituitaries not
only stimulated growth by about 10 percent but also improved the effi-
ciency of feed utilization by four percent. The only undesirable effect
noted was an increase in the fat content in certain cuts of meat. Porcine
growth hormone has been cloned and has been reported to be under devel-
opment as a commercial product (Animal Pharm, November 29, 1985, p.
18).
Two problems must be solved before any of the hormones discussed
above become an accepted tool of the animal industry. The first is that the
cost of treatment must be a relatively small fraction of the economic bene-
fit afforded by treatment. This is largely a matter of improving processes
for production and purification of hormones and the problems are no dif-
ferent in principle from those which have been solved for every antibiotic
marketed to-date. The second problem is more challenging. In all the ex-
periments above, the hormones were given as daily injections for extended
periods of time, a regimen which would probably not be acceptable to the
animal industry. What is needed is an implantable depot which releases an
appropriate quantity of hormone every day for extended periods. Several
slow release technologies are available, among which are the osmotic
pumps of the type pioneered by Alza Corporation (Palo Alto, CA), which
release hormone at a rate depending on the osmotic pressure on the vessel
containing the hormone. Also available are two types of bioerodable poly-
mer. In one type, hormone is embedded in a matrix and slowly released as
the matrix is hydrolyzed by the enzymes of the animal. In the second type,
hormone is encapsulated by bioerodable polymer and released when the
capsule ruptures. A condition approaching slow release can be achieved by
using many small capsules which have walls of different thickness. In spite
of the availability of these and doubtlessly other approaches, the develop-
ment of a convenient method to assure the controlled release of active
hormone for the extended time periods needed by the animal industry is
not a trivial problem and appears to be the only remaining obstacle to the
commercialization of the growth hormones.








IMPACTS OF CONTEMPORARY BIOTECHNOLOGY-ANIMAL SCIENCE 67

VACCINES

Bachrach (1985) reviews more than 20 potential antiviral vaccines for
animal health, all of which are based on protein antigens produced by
recombinant DNA technology. He also describes an antiprotozoal vaccine
against coccidiosis in poultry and several antibacterial vaccines. The ma-
jority of these vaccines would be impossible were it not for the ability of
recombinant DNA technology to produce the antigens in the quantities
required. In the remaining cases, traditional vaccines are either available
or potentially available but vaccines based on recombinant DNA offer the
promise of greater safety, enhanced efficacy, or reduced cost.
The vaccines described above are all of the subunitt type" which con-
tain the antigen needed to induce immunity as their only active compo-
nent. An alternative way of inducing immunity is to vaccinate with atten-
uated organisms, which are usually derived from the pathogen of interest
and can induce immunity against it, but which lack its virulence. The
Sabin polio vaccine is a classic example of a vaccine consisting of living
organisms whose virulence has been attenuated. Recombinant DNA tech-
nology can also play important roles in the development of such living
vaccines. Traditionally, attenuated organisms have been isolated by select-
ing avirulent mutants from a population of virulent parental organisms.
Even when mutagens were employed to increase the frequency of attenu-
ated mutants, the mutants were usually quite rare vis-a-vis the virulent
parental organism and their isolation required a well designed selection
and considerable persistence. Even after the attenuated mutants had been
isolated, much effort was needed to demonstrate that the mutants did not
revert to virulence, since a vaccine which could occasionally cause the
disease is obviously unacceptable. Recombinant DNA technology allows
one to introduce mutations of virtually any type into specific regions of the
genome. Thus, it is fairly simple to place a deletion, which will not revert,
into a gene known to be required by a particular pathogen for virulence.
In addition to being faster and more convenient than the traditional ap-
proach, the recombinant approach offers an important safety advantage
in that both the type of mutation and its location are known. By the tradi-
tional approach, neither the type nor location of the mutation conferring a
virulence is known, nor is it known whether mutagenesis has introduced
undesirable mutations elsewhere in the organism's genome.
The pseudorabies vaccine for swine, which has generated considerable
controversy because it is the first vaccine containing engineered organisms
to be approved by the USDA (Chemical & Engineering News, April 14,
1986, p. 4; April 21, 1986, p. 7; April 28, 1986, p. 18;New York Times,









68 R. Weppelman

April 13, 1986), provides an example of the use of recombinant DNA tech-
nology to attenuate a pathogen. In this particular case, the gene encoding
an enzyme which allows the virus to attack nerve tissue was deleted (Bio-
technology News, April 18, 1986, p. 1). Because the mutation responsible
for attenuation is a deletion, reversion to virulence is virtually impossible.


INTERFERONS AND INTERLEUKINS

Viral interference, which refers to the relative resistance of a cell al-
ready infected with virus to infection by a second virus, was first noted in
the 1930s (Hoskins, 1935; Findlay and MacCullum, 1937). The term "In-
terferon" was coined two decades later by Isaacs and Lindenmann (1957)
who demonstrated that the resistance was due to soluble factors produced
by infected cells. These factors were subsequently shown to be proteins
and three general types of interferon have been identified in humans: al-
pha, beta and gamma. The three classes are chemically distinct and tend
to be produced by different cell types (Friedman and Vogel, 1983): alpha
interferon by leucocytes; beta interferon by fibroblasts; and gamma inter-
feron by lymphocytes. Genes for all three have been cloned and, while
there appear to be only single genes for beta and gamma interferon, there
are a minimum of 14 genes for alpha interferon which vary 15 to 30 per-
cent in amino acid sequence (Friedman and Vogel, 1983; Derynck, 1983).
Whether these are alleles or "pseudogenes" (i.e., DNA sequences which
are not expressed) is not clear.
The biological actions of the interferons are broadly similar. In addition
to being antiviral, they inhibit replication of both normal and tumor cells
(Brouty-Boy6, 1980). All three classes modulate the immune response in
ways which are difficult to predict (Levy and Riley, 1983; Epstein and
Epstein, 1983; DeMaeyer-Guignard and DeMaeyer, 1985). Whether the
response is augmented or attenuated depends on the antigen, the immune
response measured, and the treatment schedule (i.e., was interferon given
before, after, or at the same time as antigen).
In view of the wide range of activities, it is not surprising that the inter-
ferons have rather serious side effects. According to Scott (1983), "It has
become apparent that interferon itself is not innocuous; indeed it was
never reasonable to consider that it would be." All interferons appear to be
pyrogenic and induce in humans a collection of side effects which can best
be described as "flu" (Scott, 1983). Growing children congenitally infected
with virus stopped gaining weight when treated with alpha interferon and









IMPACTS OF CONTEMPORARY BIOTECHNOLOGY-ANIMAL SCIENCE 69

both alpha and gamma interferons have caused weight loss in cancer pa-
tients. Whether such severe side effects will also occur in stock animals
remains to be seen.
The majority of animal health research on the interferons has been di-
rected to bovine shipping fever. This flu-like disease of the respiratory
tract, which appears to be caused by the stress associated with the transfer
of feeder calves to feed lots, costs the U.S. cattle industry an estimated
$300 to $700 million per year (Klausner, 1984). Shipping fever is also
called bovine respiratory disease complex to emphasize that a rather large
number of different pathogens, bacterial as well as viral, are commonly
involved. The start of large trials to test the efficacy of human alpha inter-
feron, bovine interferon, and an engineered consensus alpha interferon has
been announced (Klausner, 1984; Genetic Technology News, April, 1985,
p. 8). Publication of the results of these trials appears to be pending.
Interleukins 1 and 2, which are proteins that modulate the immune
response, might also be efficacious against shipping fever and other dis-
eases of stock animals. Interleukin 1, which is produced by macrophages
and certain other types of cells, stimulates T lymphocytes to produce inter-
leukin 2. Interleukin 2 in turn causes T lymphocytes to proliferate (Lach-
man and Maizel, 1980). Since T lymphocytes play several important roles
in stimulating the responses to infection, both interleukins could be thera-
peutic for a variety of diseases. Both interleukins could also be useful adju-
vants in vaccinations since they might increase the immune response if
given simultaneously with antigen. The adjuvant effect of interleukin 2
has been shown in mice by Wood et al. (1983). A gene for murine inter-
leukin 1 (Lomedico et al., 1984) and two genes for human interleukin 1
(Mosley et al., 1985) have been cloned. Bovine interleukin 2 (Genetic
Technology News, April 1986, p. 3) and a modified human interleukin 2
(Animal Pharm, October 18, 1985, p. 10) have also been cloned.
It should be cautioned that interleukin 1 has many actions in addition to
stimulating T lymphocytes to produce interleukin 2. Among these are py-
rogenicity, stimulation of various inflammatory processes, and enhance-
ment of muscle protein degradation (Gery and Lepe-Zuniga, 1984). Be-
cause of these side effects, interleukin 1, even if efficacious against various
diseases, will probably find only limited application in animal health.
Whether interleukin 2 shares these liabilities remains to be determined.









R. Weppelman


APPLICATION OF RESTRICTION FRAGMENT
LENGTH POLYMORPHISM (RFLP) ANALYSIS TO THE
BREEDING OF STOCK ANIMALS

The potential products of recombinant DNA technology discussed else-
where in this paper either contain recombinant genes or are the direct
products of recombinant genes. RFLP analysis will create no unique prod-
ucts. It has, however, the potential to accelerate conventional breeding
programs and ultimately to lead to a full understanding of the genetics of
stock animals.
Because the gene and gene products responsible for the traits valued in
stock animals are not known, breeding has always been completely empiri-
cal. One cannot tell if a particular cross has been successful until the prog-
eny are tested which in some cases can take considerable time. If the de-
sired result is a dairy bull which will sire cows that produce superior
quantities of milk, then five or more years might elapse before the success
of the cross can be determined. RFLP analysis has the potential to shorten
this time considerably. In the example of the dairy bull, one might be able
to collect his cells by amniocentesis, subject his DNA to RFLP analysis,
and thereby determine before his birth his potential as a sire of dairy cat-
tle.
RFLP analysis (Botstein et al., 1980) is based on the extremely specific
endonuclease activity of restriction enzymes. Because of their specificity,
restriction enzymes cleave DNA at a relatively small number of sites to
yield fairly long fragments. The fragments can be separated electrophoret-
ically by their size and then detected with homologous probes which have
been cloned from a genetic library. For most regions of the chromosome,
all members of a species will yield fragments of the same length. There
are, however, chromosomal regions for which members of a species are
polymorphic. Individuals which are homozygous for these regions will
yield only short or only long fragments while heterozygous individuals will
yield both short and long fragments. One can imagine that the "long"
allele lacks a cleavage site present in the short allele or alternatively that
the long allele has the same two cleavage sites but with an additional
length of DNA inserted between them. Botstein et al. (1980) have esti-
mated that 150 polymorphic sites, if appropriately located, would suffice
to analyze the entire human genome.
The utility of the polymorphic sites for animal breeding is illustrated by
returning to the example of the dairy bull. If both of the bull's parents are
heterozygous ("short plus long") for a particular site, then the progeny will
be either heterozygous "short plus long" like the parents, homozygous








IMPACTS OF CONTEMPORARY BIOTECHNOLOGY-ANIMAL SCIENCE 71

"long" or homozygous "short." The expected ratio in percent of the three
types of progeny is 50:25:25 respectively. If one knew that superior milk
production was associated with the short allele then RFLP analysis would
permit one to select the 25 percent of the male progeny which are homozy-
gous for the short allele and therefore likely to be homozygous for the genes
for superior milk production.
The major obstacle to applying RFLP analysis to stock animal breeding
is that one must know which polymorphic loci are associated with a partic-
ular trait. This involves the use of different restriction enzymes and probes
to identify a large set of polymorphic loci. These loci must then be applied
to individual animals whose pedigrees and performance traits are well-
documented to develop correlations between the loci and performance. A
substantial investment of effort would be needed but there appear to be no
major obstacles to developing lists of polymorphisms associated with par-
ticular traits. The major advantage of this approach is that it requires no
prior knowledge of the genes and gene products responsible for the traits
and it is in fact a powerful tool for identifying those genes and their prod-
ucts. The power of RFLP analysis is proven by its success in partially
mapping the human gene for Huntington's disease and in identifying carri-
ers of the gene (Gusella et al., 1983). These accomplishments are all the
more remarkable in view of the fact that absolutely nothing is known
about the primary genetic defect responsible for this neurodegenerative
disease.


HYBRIDOMA TECHNOLOGY

Hybridoma technology is based on the fusion of immortal tumor cells
with normal antibody producing cells to generate hybridomas which com-
bine in single cells the properties of antibody production and immortality.
Because hybridomas are immortal, they can produce antibody in quanti-
ties and for periods of time which are virtually unlimited. In contrast,
traditional (or polyclonall") antibodies can be obtained only in quantities
limited by the size of the animal immunized and for time periods dictated
by the animal's life span. Because all cells of a hybridoma are descendants
of a single antibody producing cell, all antibodies produced by a particular
hybridoma are "monoclonal" and are exact copies of each other: they
share a common amino acid sequence and thus bind to the same site of the
antigen with identical affinities. In contrast, polyclonal antibodies to a
particular antigen are summations of many monoclonal responses and are
thus heterogeneous. An important consequence of the homogeneity of









72 R. Weppelman

monoclonal antibodies is that monoclonals can be treated as chemical re-
agents and readily linked to other molecules to increase their utility.
The initial impact of monoclonal antibodies on animal science is in diag-
nostics and Table 1 presents a partial listing of potential diagnostics, all
based on monoclonal antibodies which have been described in the scien-
tific literature during the past two years. This list is restricted to diagnos-
tics for infections and does not include those for endocrine status, embry-
onic sex, or feed contaminants. In addition, Animal Pharm Review
(January 3, 1986, p. 14) described 24 veterinary diagnostics which were
introduced during 1985. The majority of these are based on monoclonal
antibodies and are designed to diagnose either infections or reproductive
status.

TABLE 1: Potential Diagnostics for Infectious Diseases, 1984-1986.


Organism (disease)


Reference


COMPANION ANIMALS
Dirofilaria immitis (canine heart worm)
Parvovirus (canine parvo infections)
STOCK ANIMALS
Mareks disease virus (Mareks disease
of poultry)
Avian leucosis virus (poultry leucosis)
Leptospira interrogans (bovine mastitis)

Brucella abortus (spontaneous abortions
of cattle)
Blue tongue and epizootic haemorrhagic
disease viruses (bovine B.T. & E.H.D.)
Bovine enteric cornavirus (calf scours)
Escherichia coli K99 (calf, lamb and
piglet scours)

Trichinella spiralis (swine trichinosis)
All Salmonella (contaminant of many
animal products, especially those
from poultry)


Weil et al. (1984)
Teramoto et al. (1984)

Silva and Lee (1984)

Boer and Osterhaus (1985 a,b)
Stevens et al. (1985)
Ainsworth et al. (1985)
Quinn et al. (1984)

Jochim and Jones (1984)

Crouch et al. (1984)
Holley et al. (1984)
Mills and Tietze (1984)
Morris et al. (1985)
Gamble (1984)
Mattingly (1984)


A publication by Teramato et al. (1984) describing the development of a









IMPACTS OF CONTEMPORARY BIOTECHNOLOGY-ANIMAL SCIENCE 73

diagnostic for parvovirus in dog feces illustrates the utility of monoclonal
antibodies. The authors selected two different monoclonal antibodies
which bound to different regions of the virus hemagglutination protein so
that a single antigen could simultaneously bind both types of antibody.
One of the pair served as the "capture antibody" and was bound to the
wells of microtiter plates. The second served as the "signal antibody" and
was covalently linked to the enzyme horseradish peroxidase, which can be
detected by its ability to turn a colorless substrate into an intensely colored
product. The assay consisted of three steps. First, a fecal suspension was
added to the wells containing the capture antibody, incubated to permit
any hemagglutination protein present to bind, and then removed. In this
step, the capture antibody is used to purify antigen from the fecal sample.
Second, the signal antibody was then added and incubated to permit it to
bind to hemagglutination protein which had been immobilized by the cap-
ture antibody in the first step. Unbound signal antibody was then re-
moved. Third, substrate for horseradish peroxidase was added and, after a
short incubation, color changes were noted. Those wells which turned
color contained fecal samples which were positive for parvovirus. One can
envision this assay supplied as a three component kit consisting of a solu-
tion containing signal antibody, a solution of substrate for horseradish
peroxidase, and a microtiter plate whose wells had been pre-coated with
capture antibody. Such a kit could be employed and interpreted by anyone
who could read and follow instructions. Ultimately, kits of this type will
place extremely sophisticated diagnostic tools into the hands of the average
veterinary practitioner and stockman. This will result in a reduction in the
time which lapses until appropriate therapy is initiated and should, as a
consequence, decrease losses due to mortality and morbidity. Prompt and
accurate diagnosis should also cause decreases in the inappropriate and
prophylactic uses of drugs.
Monoclonal antibodies might also find application as vaccines and other
types of biological effectors. The topography of a binding site of an anti-
body is complimentary to the topography of the antigenic site to which it
binds. It follows then that the binding site of a second antibody, which is
directed against the binding site of the first antibody, will topographically
resemble the antigen to which the first antibody binds. Second antibodies
of this type are called "anti-idiotypic" or "antiparatopic" antibodies. This
method is similar to the lost-wax process used by sculptures. A sculpted
wax object, analogous to the antigen, is used to form a ceramic mold,
analogous to the first antibody, which in turn is used to form replicas of the
object, which are analogous to the anti-idiotypic antibody. Bachrach









74 R. Weppelman
(1985) reviews several anti-idiotypic antibodies which have shown poten-
tial as vaccines.
At least in one case, monoclonal antibodies have been shown to be effec-
tive prophylactic agents (Sherman et al., 1983). Certain strains of E. coli
can colonize the intestine of neonatal calves and cause fatal diarrhea. All
of these strains have pili which are hair-like surface structures that bind to
the intestine and anchor the bacteria against peristaltic flow. If calves are
treated with monoclonal antibodies directed against pili immediately be-
fore infection, the severity of the resulting diarrhea is markedly reduced,
presumably because the antibodies prevent anchoring of the bacteria and
thereby reduce colonization.


TRANSGENIC ANIMALS

The term "Transgenic Animals" has been defined as those animals that
have stably integrated into their germ line foreign DNA which had been
introduced experimentally (Palmiter and Brinster, 1985). By this defini-
tion, the first transgenic animals were created in 1973 when the virus SV40
was introduced into the germ line of mice by injecting SV40 DNA into the
blastocoel of early embryos (Jaenisch and Mintz, 1974). However, the ex-
periment which brought transgenic animals to the forefront of scientific
imagination and public interest was the experiment by Palmiter et al.
(1982) in which mice were made transgenic with the rat growth hormone
gene. The transgenic mice not only bore the rat growth hormone gene in
their germ line but at least in some cases expressed it at high levels and
grew to a truly impressive size. The extensions of this technology to animal
production are obvious: genes could be introduced into the germ line of
breeding stock which would confer virtually any desirable trait on the
progeny including disease resistance, rapid growth, fecundity, and effi-
cient utilization of feed. Because of the germ line inheritance of the traits,
new traits could be added at the rate of a few per generation until one
obtained an animal totally customized for its intended use.
At this point, it is instructive to compare the traditional approach to
animal breeding with the transgenic approach. By the traditional ap-
proach, breeders have used the gene pool of the species, including rare
mutations, to create animals bearing the characteristics they desired. By
the transgenic approach, breeders will be able to use the collective gene
pool of all living organisms as well as beneficial mutations which they have
created to achieve the same end. Those who oppose transgenic animal
breeding because it affronts the genetic integrity of the species should note









IMPACTS OF CONTEMPORARY BIOTECHNOLOGY-ANIMAL SCIENCE 75

the qualitative similarity between transgenic and traditional breeding and
they should at the same time remember that contemporary stock animals
are the products of gene pools which have been intensively manipulated
since the dawn of domestication.
Two different approaches have been employed to produce transgenic
animals. The Palmiter/Brinster approach involves direct intervention
shortly after the egg is penetrated by sperm. At this time, the sperm's head
rounds up to form the male pronucleus which contains the genetic contri-
bution of the male to the offspring. At approximately the same time, meio-
sis is completed in the egg and the female pronucleus forms. The two
pronuclei then fuse to form a diploid nucleus which is the first nucleus of
the offspring. By the original Palmiter/Brinster approach, the recombi-
nant DNA was microinjected into the male pronucleus immediately before
fusion. Brinster et al. (1985) have subsequently shown that injection into
the female pronucleus is nearly as efficient but injection into the cytoplasm
surrounding the two pronuclei or injection into one of the two diploid
nuclei immediately after the first division is considerably less efficient.
That the DNA must be injected into the pronuclei gives rise to the only
apparent limitation of the Palmiter/Brinster approach to creating
transgenic animals: the pronuclei must be visualized to be microinjected.
The pronuclei of sheep ova could be seen only by special microscopy and
the pronuclei of swine ova were visible only after the egg's cytoplasm had
been stratified by centrifugation (Hammer et al., 1985b). These authors
found that integration of a human growth hormone gene was about 10
percent efficient in pigs but only about one percent efficient in sheep.
The second approach to producing transgenic animals employs re-
troviruses, which can be integrated into the genome of an infected cell and
inherited by progeny of that cell as a stable genetic trait. This property can
be exploited by exchanging a portion of the viral genome for the gene of
interest as was done by Jahner et al. (1985). These researchers infected
mouse embryos at the four to eight cell stage with a construct of the re-
trovirus Moloney murine leukemia virus bearing the E. coli gene for the
enzyme Xanthine (guanine) phorphoriboryl transferase. Of seven mice
which were derived from infected embryos, one male was transgenic for
the bacterial gene and his progeny were crossed to produce a strain of mice
homozygous for this locus.
In a similar experiment, Souza et al. (1984) replaced the Src gene of the
avian retrovirus, Rous Sarcomca virus, with the gene for chicken growth
hormone and then infected nine-day-old chick embryos with the recombi-
nant virus. After hatching, 50 percent of the chicks had circulating GH
levels three- to ten-fold higher than normal but did not grow more rapidly.








76 R. Weppelman
Integration into the germ line was not tested and probably did not occur
given the advanced age of the embryos when infected.
The retroviral approach to producing transgenic animals is clearly effec-
tive and in many ways more convenient than microinjection. However,
any large-scale application of the retroviral approach to producing
transgenic stock animals would result in the dissemination of genes from
viruses which as a group are associated with such diseases as AIDS and
cancer and will certainly be deferred until legitimate safety concerns have
been addressed.
A potential technical problem with transgenic animals arises from the
possibility that expression of a particular gene might be very desirable
during one phase of an animal's life but very damaging during another. As
an example, female mice transgenic for the growth hormone gene are usu-
ally infertile (Hammer et al., 1985a). This general problem doubtlessly
will be solved by including regulatory elements in the gene which put the
gene under the control of the animal's own regulatory systems or which
enable the gene to be turned on by simply adding an inducer to the ani-
mal's diet. An example of the latter is the growth hormone gene construct
used by Palmiter and Brinster to produce transgenic mice. In this construct
the growth hormone gene was placed under the control of the regulatory
element for the metallothionine gene. Because the metallothionine gene is
normally induced by zinc, growth hormone expression in the transgenic
mice could be regulated by simply adjusting the level of zinc in their diet.


SUMMARY


THE PRESENT

Biotechnology has the potential to supply the animal industry with im-
proved and novel diagnostics, vaccines, production improvers, therapeu-
tics and even stock animals. In the case of diagnostics and vaccines, this
potential has been partially realized. In the case of the growth hormones as
production improvers, realization awaits only the development of appro-
priate supporting technology by formulation scientists. And in the cases of
transgenic animals and therapeutics like the interferons and interleukins,
more research is needed to define problems and potential.









IMPACTS OF CONTEMPORARY BIOTECHNOLOGY-ANIMAL SCIENCE 77

THE FUTURE

It is indeed a tribute to recombinant DNA technology that it has, in
slightly more than a single decade, seriously depleted science's storehouse
of interesting genes to manipulate. As an example, the interferons took
centuries to be discovered, decades to be characterized, and only a few
years to be cloned. Even though the pace of science has accelerated mark-
edly since the discovery of interferons fifty years ago, it still takes far
longer to discover an important effect and to characterize the proteins and
genes responsible than it does to clone those genes. Thus, contemporary
biotechnology has given us more power over life's processes but it has not
removed the major obstacle to exercising that power which remains, as
always, our incomplete understanding of those processes. Increased under-
standing will come only from research of the most basic type and the
majority of this research must be supported by public monies and done at
universities and other nonprofit institutions. It cannot and will not be done
by the corporate sector simply because there is in this world no stock-
holder, financial analyst, or C.E.O. who has the patience to support dec-
ades of expensive research whose potential cannot even be defined. Thus,
the future progress of biotechnology will depend in a very direct way on
the extent to which public funds support basic research and the traditional
centers of basic research like universities and nonprofit institutes.
The related question concerns the type of training future scientists must
have so that they will be able to perform this basic research and exploit its
results. There have been serious comments that conventional biological
disciplines will be swallowed by the combined discipline of biochemistry/
molecular biology and equally serious comments that biochemistry/molec-
ular biology will be swallowed by the conventional disciplines. In either
case, the point is made that the biological sciences are converging at a rate
unthinkable even five years ago, and the future scientist must have inter-
disciplinary training which will enable him to conceptualize a research
problem at any intellectual level from intact animal to nitrogenous base
pair. It is a clear and difficult challenge to the university system to produce
scientists who have this breadth of training.


ACKNOWLEDGMENT

The author gratefully acknowledges the helpful comments of Dr. Dale
Bauman (Cornell University), Drs. J. Egerton, L. Gordon, H. Hafs, G.
Koo and J. Schmidt (MSDRL), and Mr. G. Barringer (MSD AGVET). The









78 R. Weppelman

inclusive literature searches of Allyson Houck and Shaun Holland of
MSDRL Information Services and the capable typing of June Zabel of
MSDRL are also gratefully acknowledged.


REFERENCES

Ainsworth, A.J., T.L. Lester and G. Capley. (1985). Monoclonal antibod-
ies to Leptospira interrogans servovar pomona. Canad. J. Comp.
Med. 49: 202-204.
Asimov, G.J. and N.K. Krouze. (1937). The lactogenic preparations from
the anterior pituitary and the increase of milk yield of cows. J.
Dairy Sci. 20: 289-298.
Bachrach, H.L. (1985). New approaches to vaccines. Adv. Vet. Sci.
Comp. Med. 30: 1-38.
Baile, C.A., M.A. Della-Ferra and C.L. McLaughlin. (1983). Perform-
ance and carcass quality of swine injected daily with bacterially-
synthesized human growth hormone. Growth 47: 225-236.
Bauman, D.E., P.J. Eppard, M.J. DeGecter and G.M. Lanza. (1985).
Responses of high-producing dairy cows to long-term treatment
with pituitary somatotrapin and recombinant somatotrapin. J.
Dairy Sci. 68: 1352-1362.
Boer, G.F. de, A.D. M.E. Osterhaus. (1985a). Application of monoclonal
antibodies in the detection of ALV-gs antigen in the Dutch avian
leucosis eradication scheme. Tijds. Diergen. 110: 842-844.
Boer, G.F. de, A.D. M.E. Osterhaus. (1985b). Application of monoclonal
antibodies in the avian leukosis virus gs-antigen elisa. Avian Path.
14: 39-55.
Botstein, D., R.L. White, M. Skolnick and R.W. Davis. (1980). Construc-
tion of a genetic linkage map in man using restriction fragment
length polymorphisms. Amer. J. Human Genet. 32: 314-331.
Brinster, R.L., H.Y. Chen, M.E. Trumbauer, M.K. Yagle and R.D. Palmi-
ter. (1985). Factors affecting the efficiency of introducing foreign
DNA into mice by microinjecting eggs. Proc. Nat. Acad. Sci. USA
82: 4438-4442.
Brouty-B6ye, D. (1980). Inhibitory effects of interferons on cell multipli-
cation. Lymphokine Rep. 1: 99-112.
Brumby, P.J. and J. Hancock. (1955). The galactopoietic role of growth
hormone in dairy cattle. New Zealand J. Sci. Tech. 36A: 447-436.
Chung, A.C.Y. and S.N. Cohen. (1974). Genomic construction between
bacterial species in vitro: Replication and expression of Staphylo-









IMPACTS OF CONTEMPORARY BIOTECHNOLOGY-ANIMAL SCIENCE 79

coccus plasmid gene in Escherichia coli. Proc. Nat. Acad. Sci. USA
71: 1030-1038.
Chung, C.S., T.D. Etherton and J.P. Wiggins. (1985). Stimulation of
swine growth by procine growth hormone. J. Animal Sci. 60: 118-
130.
Crouch, C.F., T.J.G. Raybould and S.D. Acres. (1984). Monoclonal anti-
body capture enzyme-linked immunosorbent assay for detection of
bovine enteric cornavirus. J. Clin. Micro. 19: 388-393.
DeMaeyer-Guignard, J. and E. DeMaeyer. (1985). Immunomodulation by
interferons: recent developments. Interferon 6: 69-91.
Derynck, R. (1983). More about interferon cloning. Interferon 5: 181-203.
Epstein, C.J. and L.B. Epstein. (1983). Genetic control of the response to
interferon in man and mouse. Lymphokines 8: 277-301.
Findlay, G.M. and F.O. MacCallum. (1937). An interference phenomenon
in relation to yellow fever and other viruses. J. Path. Bact. 44: 405-
424.
Friedman, R.M. and S.N. Vogel. (1983). Interferons with special emphasis
on the immune system. Adv. Immun. 34: 97-140.
Gamble, H.R. (1984). Application of hybridoma technology to the devel-
opment of a diagnostic test for swine trichinosis. In Hybridoma
Technology in Agriculture and Veterinary Research pp. 274-281.
Gery, I. and J.L. Lepe-Zuniga. (1984). Interleukin 1: uniqueness of its
production and spectrum of activities. Lymphokines 9: 109-125.
Gusella, J.F., N.S. Wexler, P.M. Coneally, S.L. Naylor, M.A. Anderson,
R.E. Tanzi, P.C. Watkins, K. Ottina, M.R. Wallace, A.Y. Saka-
guchi, A.B. Young, I. Shoulson, E. Bonilla and J.B. Martin. (1983).
A Polymophic DNA marker genetically linked to Huntington's dis-
ease. Nature 306: 234-238.
Hammer, R.E., R.L. Brinster and R.D. Palmiter. (1985a). Use of gene
transfer to increase animal growth. Cold Spring Harbor Symp.
Quant. Biol. 50: 379-387.
Hammer, R.E., V.G. Pursel, C.E. Roxroad Jr., R.J. Wall, D.J. Bolt, K.M.
Ebert, R.D. Palmiter and R.L. Brinster. (1985b). Production of
transgenic rabbits, sheep and pigs by microinjection. Nature 315:
680-683.
Holley, D.L., S.D. Allen, B.B. Barnett. (1984). Enzyme-linked immuno-
sorbent assay, using monoclonal antibody, to detect enterotoxic Es-
cherichia coli K99 antigen in feces of dairy calves. Amer. J. Vet.
Res. 45: 2613-2616.
Hoskins, M. (1935). A protective action of neurotropic against viscerotro-









80 R. Weppelman

pic yellow fever virus in macacus rhesus. Amer. J. Trop. Med. Hyg.
15: 675-680.
Isaacs, A. and J. Lindenmann. (1957). Virus interference I: The inter-
feron. Proc. Royal Soc. B147: 258-267.
Jaenisch, R. and B. Mintz. (1974). Simian virus 40 DNA sequences in DNA
of healthy adult mice derived from preimplantation blastocysts in-
jected with viral DNA. Proc. Nat. Acad. Sci. USA 71: 1250-1254.
Jihner, D.K. Haase, R. Mulligan and R. Jaenisch. (1985). Insertion of the
bacterial gpt gene into the germ line of mice by retroviral infection.
Proc. Nat. Acad. Sci. USA 82: 6927-6931.
Jochim, M.M. and S.C. Jones. (1984). Identification of B.T. and EHD
viruses by immunofluorescence with monoclonal antibodies. Proc.
Ann. Meet. American Association of Veterinary Laboratory Diag-
nosticians 26: 277-286.
Johnsson, I.D. and I.C. Hart. (1985). The effects of exogenous bovine
growth hormone and bromocryptine on growth, body develop-
ment, fleece weight and plasma concentrations of growth hor-
mone, insulin and prolactin in female lambs. Animal Production
41: 207-217.
Klausner, A. (1984). IML introduces interferon product for cattle. Bio/
Technology October 1984, 841.
Kohler, G. and C. Milstein. (1975). Continuous cultures of fused cells se-
creting antibody of predefined specificity. Nature 256: 495-497.
Lachman, L.B. and A.L. Maizel. (1980). Human immunoregulatory mol-
ecules: interleukin 1, interleukin 2 and B-cell growth factor. Con-
temp. Topics Molec. Immun. 9: 147-167.
Levy, H.B. and F.L. Riley. (1983). A comparison of immune modulating
effects of interferon and interferon inducers. Lymphokines 8: 303-
322.
Lomedico, P.T., V. Gubler, C.P. Hellman, M. Dukovitch, J.G. Giri, Y-CE.
Pan and K. Collier. (1984). Cloning and expression of murine inter-
leukin 1 cDNA in E. coli. Nature 312: 458-462.
Maniatis, T., E.F. Fritsch and J. Sambrook. (1982). Molecular Cloning, A
Laboratory Manual Cold Spring Harbor Laboratory, Cold Spring
Harbor, NY.
Mattingly, J.A. (1984). An enzyme immunoassay for the detection of all
Salmonella using a combination of a myeloma protein and a hybri-
doma antibody. J. Immun. Meth. 73: 147-156.
Mills, K.W. and K.L. Tietze. (1984). Monoclonal antibody enzyme-linked
immunosorbent assay for identification of K99-positive Escherichia
coli isolates from calves. i. Clin. Micro. 19: 498-501.









IMPACTS OF CONTEMPORARY BIOTECHNOLOGY-ANIMAL SCIENCE 81

Morris, J.A., C.J. Thorns and C. Boarer. (1985). Evaluation of a mono-
clonal antibody to the K99 fimbrial adhesin produced by Escheri-
chia coli enterotoxigenic for calves, lambs and piglets. Res. Veter.
Sci. 39: 75-79.
Mosley, B., C.J. March, A. Larsen, D.P. Cerretti, G. Braedt, V. Price, S.
Gillis, C.S. Henney, S. Kronheim, K. Grabstein, P.J. Conlon, T.P.
Hopp and D. Cosman. (1985). The cloning and expression of two
distinct human interleukin-1 (IL-1) cDNAs. In: The physiological,
metabolic and immunologic actions of interleukin-1 Alan R. Liss,
NY. pp. 521-532.
Muir, L.A., S. Wien, P.F. Duquette, E.L. Rickes and E.H. Cordes. (1983).
Effects of exogenous growth hormone and diethylstilbestrol on
growth and carcass composition of growing lambs. J. Animal Sci.
56: 1315-1323.
Palmiter, R.D. and R.L. Brinster. (1985). Transgenic mice. Cell 41: 343-
345.
Palmiter, R.D., R.L. Brinster, R.E. Hammer, M.E. Trumbauer, M.G. Ro-
senfeld, N.C. Birnberg and R.M. Evans. (1982). Dramatic growth
of mice that develop from eggs microinjected with metallothionein-
growth hormone fusion genes. Nature 300: 611-615.
Quinn, R., A.M. Campbell and A.P. Phillips. (1984). A monoclonal anti-
body specific for the A antigen of Brucella spp. J. Gen. Micro. 130:
2285-2289.
Rodriquez, R.L. and R.C. Tait. (1983). Recombinant DNA Techniques:
An Introduction published by: Addison-Wesley Publishing Co.,
Reading, MA.
Scott, G.M. (1983). The toxic effects of interferon in man. Interferon 5:
85-114.
Sherman, D.M., S.D. Acres, P.L. Sadowski, J.A. Springer, B. Bray, T.J.G.
Raybould and C.C. Muscoplat. (1983). Protection of calves against
fatal enteric colibacillosis by orally administered Escherichia coli
K99-specific monoclonal antibody. Infection and Immunity 42:
653-658.
Silva, R.F. and L.F. Lee. (1984). Monoclonal antibody-mediated immuno-
precipitation of proteins from cells infected with Marek's disease
virus or turkey herpes virus. Virology 136: 307-320.
Souza, L.M., T.C. Boone, D. Murdock, K. Langley, J. Wypych, D. Fen-
ton, S. Johnson, P.H. Lai, R. Everett, R-Y. Hsu and R. Bosselman.
(1984). Application of recombinant DNA technologies to studies on
chicken growth hormone. J. Exp. Zool. 232: 465-473.
Stevens, A.E., S.A. Headlam, D.G. Pritchard, C.J. Thorns and J.A. Mor-









82 R. Weppelman

ris. (1985). Monoclonal antibodies for diagnosis of infection with
Leptospira interrogans servovor hardjo by immunofluorescence.
Veter. Rec. 116: 593-594.
Sutherland, S. (1985). An enzyme linked immunosorbent assay for detec-
tion of Brucella abortus in cattle using monoclonal antibodies.
Aust. Veter. 1. 62: 264-268.
Teramoto, Y.A., M.M. Mildbrand, J. Carlson, J.K. Collins and S. Win-
ston. (1984). Comparison of enzyme-linked immunosorbent assay,
DNA hybridization, hemagglutination, and electron microscopy
for detection of canine parvovirus infections. J. Clin. Micro. 20:
373-378.
Weil, G.J., M.S. Malane, K.G. Powers and L.S. Blair. (1984). Monoclonal
antibody-based assay for parasite antigenemia in Dirofilaria im-
mitis-infected dogs. Fed. Proc. 43: 1667.
Wood, D.D., M.J. Staruch, PL. Durette, W.V. Melvin III and B.K. Gra-
ham. (1983). In: Interleukins, Lymphokines and Cytokines Aca-
demic Press, NY. pp. 691-696.
Young, F.G. (1947). Experimental stimulation (galactopoiesis) of lacta-
tion. Brit. Med. Bull. 5: 155-160.








PART III
ADVANCES IN
BIOTECHNOLOGY:
ENVIRONMENT, HEALTH
AND ENERGY













Impacts of Biotechnology
on Biomedical Sciences

Thomas W. O'Brien
Department of Biochemistry and Molecular Biology
University of Florida
Gainesville, Florida

Researchers working in the biomedical sciences are well aware of the
tremendous positive impact that biotechnology has had in this area. As
illustrated in Figure 1, much of the biotechnology related research leading
to medical advances involves recombinant DNA approaches, the produc-
tion and utilization of monoclonal antibodies, and most recently, protein
engineering. A most fortunate aspect of this research is that its very prod-
ucts have a direct positive feedback on research capabilities, providing
new tools to carry out this kind of research. The rapid expansion of recom-
binant DNA methodologies following the introduction of DNA restriction



RESEARCH


APPROACHES PRODUCTS MEDICAL ADVANCES

Recombinant DNA Diagnostics Molecular Basis
Monoclonal Antibodies Therapeutics of Disease
Protein Engineering Gene Therapy Human Genes
Mapping
Structure
Regulation

Figure 1. Impacts of biotechnology on the biomedical sciences. Medical
advances and medical products of biotechnology are shown in relation to
biotechnology research, emphasizing the possible feedback of the inter-
mediate products of biotechnology on research in this area.









T. O'Brien


enzymes is a good example of this process whereby early advances and
products led to the development of new vectors and new research tools.
This, in turn, speeds the discovery process.
Important medical advances are being made as a result of this rapid
enhancement of biomedical research through biotechnology. With each
advance comes increased understanding of the molecular basis of disease.
Along these lines, encouraging progress is being made in the study of the
human genome. Increased numbers of human genes are being identified
and localized at high resolution on individual chromosomes. Finally, given
knowledge of their localization and detailed structure, we will be in a
better position to understand their regulation. Despite the optimism of the
moment, it should be emphasized that we are only in the beginning stages
of what promises to be a long and interesting endeavor.
The major products of biotechnology in the biomedical area fall into the
broad categories of diagnostics and therapeutics. We shall consider these in
turn, ending with the topic of gene therapy.


DIAGNOSTICS

The biotechnology-derived diagnostics are of two main kinds, mono-
clonal antibodies and DNA probes. Several companies, including Cento-
cor, Cetus, Damon Biotech and Monoclonal Antibodies, among others,
have major involvements in the production of monoclonal antibodies for
diagnostic purposes. Over 150 different monoclonal antibodies are on the
market now. Mainly used for diagnosis of various infectious diseases, they
are also being used to detect various cancers and for pregnancy testing.
Because of their specificity and the constancy of their properties, mono-
clonal antibodies are ideally suited for several diagnostic applications. One
of the real advantages of monoclonal antibodies, in addition to those dis-
cribed by Weppelman (this volume), is the fact that they are indeed de-
rived from an individual clone. Thus, for example, hybridomas producing
antibodies against cancerous cells can be cloned and screened to identify
those producing antibodies against cell surface determinants that are
found only on the cancerous cells, and not on normal cells. Such mono-
clonal antibodies, specific for different kinds of cancer cells, are now being
produced not only for the detection and identification of different kinds of
cancerous cells, but also for the localization of specific tumors. A recent
example of the latter application involves the noninvasive imaging of ovar-
ian carcinoma using monoclonal antibodies produced against the carci-
noma-specific determinants by Centocor. Such antibodies can be tagged




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