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Foderal Register / Vol. 46, No. 210 / Friday, October 30, 1981 / Notices 
Appendix A on the baeis that they 
exchange genetic information by known 
physiological processes. The second 
part of Dr. McKay’s proposal involves 
permission for the transfer of a 
recombinant plasmid from 5. faecalis to 
S, iacth. 
This proposal was published in the 
Federal Regteter of August 4, 19S1 (46 FR 
39770), and no comments were received 
on the proposal. 
The RAC recommended approval of 
these requests by a vote of fourteen in 
favor, none opposed, with no 
abstentions. 
I accept this recommendation. 
Streptococcus lactis has been added to. 
Sublist E, Appendix A, as follows: 
"Sublist E. 
“One way transfer of Streptococcus 
mutans or Streptococcus lactis DNA 
into Streptococcus sanguis. " 
A new entry, item 37, is added to 
Appendix E as follows: 
“37. Permission is given to transfer a 
recombinant lactose plasmid from 
Streptococcus faecalis io S. lactis by 
conjugation.” 
G. Cloning of Saccharomyces 
Cerevisiae DNA in Salmonella 
Typhimurium 
Drs. Christopher Marvel emd Edward 
Penhoet of the University of California, 
Berkeley, in a letter dated July 8, 1981, 
requested permission to clone 
Saccharomyces cerevisiae DNA in 
Salmonella typhimurium using the 
plasmid, YEpl3. The principal 
investigators wish to utilize an 
attenuated laboratory strain of 
Salmonella typhimurium to screen for 
the yeast pseudouridine synthetase 
gene. After screening, the selected 
plasmids will be maintained in an E. coli 
host. 
A Federal Register announcement of 
the proposed experiments appeared on 
August 4, 1981 (46 FR 39771). During the 
comment period, no comments were 
received. 
The RAC discussed this proposal at 
the September 10-11, 1981 meeting. As 
the Salmonella strains to be employed 
are attenuated laboratory strains, it was 
suggested that Pi physical containment 
conditions are adequate. By a vote of 
fourteen in favor, none opposed with no 
abstentions, RAC recommended that the 
experiments be permitted at the PI level 
of containment. 
I accept this recommendation and a 
new item, number 38. has been added to 
Appendix £: 
“38. Attenuated laboratory strains of 
Salmonella typhimurium may be used 
under Pi physical containment 
conditions to screen for the 
Saccharomyces cerevisiae 
pseudouridine synthetase gene. The 
plasmid YEpl3 will be employed as the 
vector.” 
H. Request To Utilize Haemophilus 
Parainfluenzae to Clone Moloney 
Murine Leukemia Pro virus 
The RAC at its September 16-11, 1961, 
meeting considered a proposal from Dr. 
James Gautsch of the Sciipps Clinic and 
Research Foundation, La Jolla, 
California, to introduce Moloney MuLV 
provirul DNA and flanking mouse DNA 
into Haemophilus parainfluenzae 
employing a plasmid cloning vector with 
a broad host range in gram negative 
bacteria. The recombinant viral DNA 
would be subsequently used to transfect 
mouse cells in culture. Notice of this 
request appeared in the Federal Register 
of August 4, 1981 (46 FR 39770). During 
the comment period, no responses wele 
received. 
The RAC discussed this request at its 
September 16-11, 1981 meeting. It was 
noted that H. parainfluenzae is not 
considered a pathogen. Also, Moloney- 
MuLV virus can only infect mouse or rat 
cells, and not human cells; the virus is 
classiHed as a low-risk oncogenic virus. 
The RAC by a vote of eleven in favor, 
none opposed, with three abstentions 
recommended approval of the proposed 
experiments at ^e P2 level of physical 
containment. 
I accept this recommendation and text 
has been added to Appendix E of the 
Guidelines, as follows: 
“39. Permission is granted to clone in 
Haemophilus parainfluenzae Moloney 
murine leukemia provirus and mouse 
cellular flanking sequences employing 
the plasmid cloning vector, pRK290, 
under P2 containment conditions.” 
I. Request for Development of a New 
Host- Vector System Based on^ 
Corynebaderium Glutamicum 
A request of Dr. Daniel Liberman, 
Massachusetts Institute of Technology, 
for an evaluation of the containment 
conditions for development of a new 
host-vector system based on a gram 
positive bacterium Corynebacterium 
glutamicum as host was considered by 
the RAC at its September 10-11, 1981 
meeting. A notice of the proposal 
appeared in the Federal Register of 
August 4, 1981. No comments were 
received on the proposal. 
Three possible types of vectors, 
including hybrid plasmids were 
proposed to be used with the new 
cloning host. It was noted during the 
RAC discussion that this organism is a 
nonpathogen and is employed in the 
conunercial production of amino acids. 
A motion to approve the request at the 
Pi level of contaiiunent provided that 
non-conjugative poorly mobilizable 
plasmids are used was passed by a vote 
of eleven in favor, one opposed, with 
one abstention. 
1 accept this recommendation and text 
has been added to Appendix E of the 
Guidelines. 
J. Use of Confugative Plasmids To 
Transfer DNA Between Escherichia 
Coli, Vibrio Cholera, and Vibrio 
Harveyi 
Dr. J. W, Hastings of Harvard 
University requested permission to 
move vibrio harveyi DNA cloned in 
Escherichia coli K-12 to Vibrio cholera 
and then to Vibrio harveyi. The 
investigator hopes to develop a system 
of genetic manipulation in V. harveyi in 
order to clone the luminescence gene(s). 
A notice of this proposal appeared la 
the Federal Register of August 4, 1981 
(46 FR 39771), for public comment 
During the comment period, no 
comments were received. 
The RAC discussed the proposal at 
the September 10-11, 1981 meeting. At 
the meeting, it was noted that the 
investigators hoped to enrich for an 
intermediate “donor” strain (V. cholera) 
which, under standard culture 
conditions, can mobilize plasmid DNA 
into V. harveyi at a significantly higher 
frequency than can be obtained by 
direct mating of V. harveyi with E. coil. 
As Vibrio cholera is a CDC class 2 
organism, RAC felt it was appropriate to 
require use of P2 physical containment 
conditions for those experiments 
utilizing V. cholera as die intermediate 
“donor” strain. Hie other described 
experiments could proceed under PI 
containment conditions. By a vote of ten 
in favor, none opposed, with three 
abstentions, RAC recommended that the 
requests be approved. 
I accept this recommendation. A new 
item, number 41, has been added to 
Appendix E as follows: 
“41. Vibrio harveyi DNA may be 
cloned in Vibrio cholera; plasmids may 
be used to transfer the cloned V. 
harveyi DNA between E. coli K-12, V, 
cholera and V. harveyi. P2 physical 
containment conditions are required for 
those experiments involving V. cholera. 
PI containment conditions may be used 
for other phases of the project.” 
K. Containment Conditions for Cloning 
and Expression of DNA coding for 
Diphtheria Toxin 
At the April 1981 RAC meeting, Dr^ 
John Murphy of Harvard University was 
given permission to clone in E. coli K- 
12, under P4 containment conditions, 
restriction fragments of Corynephage 
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