December 27, 1950

Proposals to the Atomic Energy Commission for a Contract in

aid of a Research Project on
CYTOGENETIC EFFECTS OF RADIATIONS ON BACTFRIA

at the Depertment of Genetice, UNIVERSITY OF WISCONSIRZ.

Project Leader: Joshua Lederberg, Ph.D., Associate Professor of Genetics

Scope, status, and motivation of projest:

Support is reouested from the Atomic Energy Commission for the pre-
liminary exploration of a subject which may throw fundanentel light on
the mechaniam of biological action of ionising radistions. The material
to be studied is diploid cells of Escherichis Soli. Work in progress has
shown that this material cen be used to test the significance of recessive
lethal mutations in the bactericidal effects ef chemical mutagens such as
nitrogen sustard or physical agents such as ultra-violet light. In diploid
cella, lethal autetions can be detected wing to their temporary masking
by the unmutated genome; in haploid sells, of coures, lethal mutetions
Feanlt in cell death, end ,re thus not amenable to study.

Work in progress indicates th=t, contrary to expectation, lethal
gutations play an insignificant role in bactartal Killing by nitrogen
mustard or ultraviolet light. FBactariaide by these, and a large number
of other agents, including hylrogen peroxide, formeldehyde, end X-rays
(in a single preliminary experiment) is, however, accompanied by complex

Genetic peorganlzations, not completely understood as yet,  sut including
the production of cells with haploid, rather than diploid constitution.
It is also samurent that these changes are accompanies? by 1! aturhancas
in the division and distribution of tho anelosr bodies tn the treated
pacteria.

The present project concerns the =xtensgion of this genetic and
eytologizsul vork with speciz] reference to the effects of X-rays (and
subsequently, of ether donising ss tiatione). Beeause the vroject is
already under way to some extent, with respect to chemical and ultra-
vielet 2ffects, 2 modest request is made for the firet year, as the major
aim of the project iuring wiis interval is te determine the advisabliity
of a concerted attack upon this problem. It is snticipated thet increased
working epace for rasesroh vill be available to the investigetor in 1952,
if an expeneion of the program in the areaa of interest te the Atomic
Energy Comuissiloa appears desirable, both) the Commission end the investicator.

In this interval ef explerstion, probebly no new personnel will be
requires’, The genetic work will be ccatinved by the investigetor, with
the assistance, partionlarly in cytological aspects, of graduate student
research assistants already in reeidence or possibly of an additional
assist. nt later. Pinancial support will be required primarily for the
purchase of roentgen-dosimetric equipment needed for kinetic studies
on the comparative rates of sterilization of diploid and haploid cells,
and fer additicusl supplied incidental to the project.

The University of Wisconsin hae « number of X-rey machines on the
campus, although none eof these is primarily suited to biologicel research
However, for the present, the rorchase of a new machine for the evrrent
project dees not seem to be justified, and it is hoped that the presant

facilities will be adequate, A radioactive Co source will be avsilable
to a project in ancther research group, and the installation of a detatron
40 anticipated. Basic apparatus and facilities for bacteriological research

have already been established.

Relevant literature:

Lederberg, J. 1947 Gene recombination and linked segregations in
Escherichia eoli. Genetics 32: 505-525.

1949 Aberrant heteresygotes in Eecherichia coli. Proc.
Hat. Acad. Bei. 35:178~-84.

1951 Genetic experiments on beeteria. In "Fifty Years
of Genetics." MacMillan: New York, in press.

 

Delaporte, B. 1949 Cytology of Bacteria. Carnegie Inst. Irbk. 46:
166-170.

Bewcombe, B. B. and Scott, 9. W. 1949 Factors responsible for the

delayed appearance of radiation~induced mutants in Escherichia
coll, Genetics 34:475-492.

Scientific Personnel: In this preliminary period, no new scientific
personnel are contemplated, and the project will be handled directly
by the principal investigator, with the essistance of various graduate
etudents already in residence. If necessary, an sdditional graduate
assistant may be employed during the latter half of the contract tern,
but no specific person ie under consideration,

Proposed budget. (For March 1, 1951 to Februery 29, 1952)

The University of Wisconein will provide basic working feeilitias
for bacteriological research. Because the proposed project interlocks
with other work already in progress, glassware, reagents, and bacteriological
media will also be provided, as well es the services of the principal
investigator. X-ray equipment already on the campus will be used as
required, but fees routinely charged in connection with certain such

services when provided by other departments will be paid from the contract.
fhe Commisaion will be asked to provide for the following:

Hquipment and Supplies; 350

Roentgen meter for calibration of X-ray doses 250

Accessories for phetomicrography 100
Service Fees for X-ray equipment 50
Salary, for 1/2 time graduate assistant for 6 months 600

Total “1000
Overhead, at 5%, ——-80
Grand Tctal 1989

fhe proposed term for the preliminary project in Mareh 1, 1951 to
February 29, 1952. However, if reseach results or other factors inter-
yene which make further action advisable, the investigator may apply
for subsidiary support prior to the expiration of this term. The
probable term of an extended project would be about four years, unless
full-time employees displace our graduate students.
Responsibilities of investigater: The principal iavestigater is in
charge of allaboratory devoted to microbial genetic research, and housing,
at present, three graduate research assistants, ons postdoctoral research
associate and a number of undergraduate assistants. His teaching duties,
aside from those incidental to the laboratory, are minimal, A number of
research vrojects are unier way in the laboratory, dealing primarily with
(a) formal and physiological genetics of Escherichia coli; (b) host-
parasite relationships in lysogenic bacteria; (c) screening for genetic
recombination in Salmonella and other bacteria; (da) cytological studies
of haploid and diploid E. celi, and (eo) mechaniam of action of bactericidal
agents,

The laboratory's program receives part of its support from other federal

agencies as follows:
For 1950-51! Metionel Institutes of Health $4 320
(Genetics of Salmonella)

Chemicel Corps 6,000 (15 mos.)
(Lysogenic Becteria)

Requests anticipated for 1951-52

Betione) Inatitutes of Health 9,000
(Genetics of Braeterta)

eo et eee nee eee oreaeoneenreaeneneane