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VII. MEMBERS OP THE RECOMBINANT DNA MOLE- 
CULE PROGRAM ADVISORY COMMITTEE 
Chairman 
Stetten, DeWltt, Jr., M.D., Ph.D., Deputy Di- 
rector for Science, National Institutes of 
Health. 
Vice Chairman 
Jacobs, Leon, Ph.D., Associate Director for 
Collaborative Research, National Institutes 
of Health. 
Adelberg, Edward A., Ph.D., Professor, De- 
partment of Human Genetics, School of 
Medicine, Yale University. 
Chu, Ernest H. Y., Ph.D., Professor, Depart- 
ment of Human Genetics, Medical School, 
University of Michigan. 
Curtiss, Roy, III, Ph.D., Professor, Depart- 
ment of Microbiology, School of Medicine, 
University of Alabama. 
Darnell, James E., Jr., M.D., Professor, De- 
partment of Molecular Cell Biology, Rocke- 
feller University. 
Hellnskl, Donald B., Ph.D., Professor, De- 
partment of Biology, University of Cali- 
fornia, San Diego. 
Hogness, David S., Ph.D., Professor, Depart- 
ment of Biochemistry, Stanford University. 
Kutter, Elizabeth M., Ph.D., Member of the 
Faculty, In Biophysics, The Evergreen State 
College. 
Littlefield, John W., M.D., Professor & Chair- 
man, Department of Pediatrics, Children’s 
Medical & Surgical Center, Johns_^Hopklns 
Hospital. 
Bedford, Emmette S., Ph.D., LL.D., Ashbel 
Smith Professor of Government and Public 
Affairs, Lyndon B. Johnson School of Pub- 
lic Affairs, University of Texas at Austin. 
Bowe, Wallace P., M.D., Chief, Laboratory of 
Viral Diseases, National Institute of Al- 
lergy & Infectious Diseases, National In- 
stitutes of Health. 
Setlow, Jane K., Ph.D., Biologist, Brook- 
haven National Laboratory. 
Splzlzen, John, Ph.D., Member and Chair- 
man, Department of Microbiology, Scrlpps 
Clinic & Besearch Foundation. 
Szybalskl, Waclaw, D.Sc., Professor of On- 
cology, McArdle Laboratory, University of 
Wisconsin. 
Thomas, Charles A., Jr., Ph JJ., Professor, De- 
partment of Biological Chemistry, Harvard 
Medical BchooL 
Executive Secretary 
Gartland, William J., Jr., PhJ3., Health Sci- 
entist Administrator, National Institute of 
General Medical Sciences, National Insti- 
tutes of Health. 
Liaison Representatives 
Hedrlch, Richard, Ph.D., Coordination Pro- 
gram of Science Technology & Human Val- 
ue, National Endowment for the Humani- 
ties. 
Lewis, Herman W., Ph.D., Division of Biologi- 
cal and Medical Sciences, National Science 
Foundation. 
Nightingale, Elena O., Ph.D., Assembly of 
Life Sciences, National Academy of Sci- 
ences. 
Shepherd, George R., PhJ)., Division of Bio- 
medical and Environmental Research, 
Energy Research and Development Ad- 
ministration. 
Appendix A to Appendix D 
BTATEMENT ON THK T7SE OP BACILLUS SUBTILIS 
IN XECOMBINANT AIOLECULK TECHNOLOGY 
Unquestionably, Escherichia coli Is the 
most well characterized unicellular orgahism. 
Years of basic research have enabled Investi- 
gators to develop a well characterized genetic 
m^p, to obtain detailed knowledge of virulent 
arid temperate bacteriophages, and to explore 
the physiology, genetics, and regulation of 
plasmids. More recently, the development of 
DNA-medlated transformation has permitted 
exogenous fragments or molecules of DNA 
to be Incorporated Into the genome or to 
reside as self-repllcatlng units. The dis- 
covery of transformation of Bacillus subtilis 
by Splzien (1) stimulated the development 
of analternatlve model system. The purpose 
of this report Is to summarize the current 
status of this genetic system and to describe 
the actual and potential vectors and vehicles 
available . for recombinant molecule tech- 
nology. 
A. CURRENT KNOWLEGDE OF THE CHROMOSOMAL 
ARCHITECTURE AND MECHANISMS OF GENETIC 
EXCHANGE IN B. “SUBTILLS" 
Two mechanisms of genetic exchange have 
been utilized to establish the linkage map of 
B. subtilis, DNA-medlated transformation 
(capable of transferring approximately 1% of 
the genome) and transduction with bacterio- 
phage PBSI (capable of transferring 6-8% of 
the chromosome) . Recent detailed genetic 
studies with PBSI by Lepesant-Kejzlorovk et 
al. (2) have resulted In the development of 
a circular genetic map for this organism. 
The current edition of the map (3) contains 
196 loci. Biophysical analyses have estab- 
lished .that the chromosome Is circular (4) 
and replicates bidirectionally (6). 
Transformation with purified fragments of 
DNA Is a highly efiSclent process In B. subtilis 
with frequencies of 1 to 4% usually attained 
for any auxothrophlc or antibiotic resistance 
markers. Frequencies of approximately 10% 
transformation can be achieved with DNA 
prepared from gently lysed L-forms or pro- 
toplasts (6). These large fragments of DNA 
are readily Incorporated by the recipient celL 
Generallzed transduction occurs with bac- 
teriophages SPIO (7), PBSI (8), and SPPl 
(9), while a low frequency of specialized 
transduction has been reported with bac- 
teriophage ^105 (10). 
Although transformation Is most efficient 
In homologous crosses (B. subtilis Into B. 
subtilis) , It has also been possible to exchange 
DNA among closely related species (11). The 
most extensively studied members of the B. 
subtilis genospecles Include B. licheniformis, 
B. pumilus, B. amyloliquef Odens, and B. 
globigii (refer to reference 12 for a review and 
references 13-16 for examples of this heterol- 
ogous exchange) . This exchange occurs even 
