Federal Register / Vol. 46, No. 126 / Wednesday, July 1, 1981 / Notices 
34459 
in all experiments for which HV2 N. crassa 
systems are approved, provided that only 
DNA from Class 1 agents is used. 
In the submission. Dr. Perkins argued 
that present containment levels cannot 
be iustiHed by any demonstrated 
hazard, especially when only Class 1 
agents are involved, and that present 
containment levels impose a serious and 
unnecessary handicap on genetic 
analysis of Neurospora. 
Three letters were received strongly 
supporting this proposal. One 
commentator said that it was his 
experience that contamination by 
airborne Neurospora conidia is not a 
serious problem. Another commentator 
stated; 
. . . Since Neurospora crassa is not a pest 
or pathogen, there is no jusliTication for 
imposing more stringent containment 
conditions for research with Neurospora than 
for investigations with yeast or animal 
viruses. The current Guidelines are seriously 
hindering the use and development of 
Neurospora as a subject for recombinant 
DNA research. 
The RAC recommended approval of 
the proposal by a vote of 10 in favor, 0 
opposed, with 5 abstentions. 
I accept this recommendation, and 
entry 2 of Appendix E has been 
modified to reflect this decision. 
IX. Request to Employ a Conjugative 
Plasmid To Transfer Neurospora Crassa 
DNA 
Dr. Norman Giles of the University of 
Georgia in a letter dated March 18. 1981, 
requested permission to use a 
conjugative plasmid to transfer the qa-2 
gene of Neurospora crassa among E. 
coU K-12 strains. The N. crassa qa-2 
gene would be ligated into a derivative 
(pVK57) of the mobilized plasmid 
RSF2124. 
In support of his proposal. Dr. Giles 
argued that (1) no Neurospora protein 
other than the qa-2 protein (catabolic 
dehydroquinase) is produced in E. coU 
transformed with plasmid pVK57; no 
hybrid or non-functional proteins are 
produced, and (2) the catabolic 
dehydroquinase synthesized by 
Neurospora is identical to that 
synthesized by E. coli, based on 
heatstability, sedimentation, and amino 
acid composition. In addition, fragments 
close to the NHi-terminal end have been 
sequenced and are identical in E. coli 
and Neurospora crassa. 
A Federal Register announcement of 
Dr. Gilles' request appeared on March 
20. 1981 (46 FR 17997). During the thirty 
day comment period, no comiYients were 
received. 
The RAC discussed the request at its 
April 23-24, 1981 meeting. Noting that 
the qa-2 fragment was relatively well- 
defined. RAC recommended approval of 
the proposal by a vote of 17 in favor, 0 
opposed, with no absentions. P2 
containment conditions were specified. 
I accept this recommendation. A new 
entry, number 33. has been added to 
Appendix E as follows: 
A conjugative plasmid may be used to 
transfer among E. coli K-12 strains at P2 
physical containment the qa-2 gene oT 
Neurospora crassa ligated to a mobilizable 
plasmid. 
X. Request To Use An E. Coli Strain 
Containing Mu Phage Insertions 
Dr. Darold Molten of the University of 
California at Riverside, in a letter dated 
March 18. 1981, requested permission to 
utilize the E. coli strain DF214 (or 
derivatives thereof), and plasmid 
vectors (e.g.. pBR322, pBR325) to clone 
rat cDNA. Strain DF214. a K-12 
derivative.contains (1) a Mu phage 
insertion in the phosphoglucose 
isomerase gene, and (2) a Mu lysogen in 
an unknown location. Dr. Molten 
calculates the frequency of Mu lysis and 
transduction in DF214 to be of the order 
of 10-'* to 10-'*. 
The Federal Register of March 20, 1981 
(46 FR 17997) carried an announcement 
of Dr. Molten's proposal. No comments 
were received during the thirty day 
comment period. 
The RAC discussed the proposal at 
the April 23-24, 1981 meeting and 
recommended by a vote of 17 in favor, 0 
opposed, with no absentions. that the 
initial screening of the rat library be 
done at P2 containment; after the clone 
of interest has been purified, it may be 
worked with at Pi containment. 
I accept this recommendation, and a 
new entry, item 34, will be added to 
Appendix E as follows: 
E. coli K-12 strain DF214 (or derivatives 
thereof) and plasmid vectors (e.g., pBR322. 
pBR325) may be used to clone rat cONA 
under P2 conditions. After the clone of 
interest has been purified, it may be worked 
with under Pi containment. 
Additional Announcements of The 
Director, NIH 
Section IV-E-l-b-(3)-(d) of the 
Guidelines gives responsibility to the 
Director, NIM, for “authorizing, under 
procedures specified by the RAG. large- 
scale experiments (i.e., involving more 
than 10 liters of culture) for recombinant 
DNAs that are rigorously characterized 
and free of harmful sequences." 
Accordingly, several requests for 
authorization to culture, on a large- 
scale. recombinant DNA host-vector 
systems have been received and 
reviewed by the NIM. 
I. Professor Barry T. Nall 
On June 16, 1981, the Director, NIM, on 
the recommendation of the RAC, 
approved a request from Professor Barry 
T. Nall, University of Texas Mealth 
Science Center at Mouston, Mouston, 
Texas 77025, for the large-scale culture 
of EKl host-vector systems into which 
have been ligated recombinant DNA 
plasmids containing E. coli DNA and 
Saccharomyces cerevisiae genes for 
cytochrome c. 
The principal investigator is Dr. Nall. 
The work is to be done at the Pl-LS 
level. 
///. Cetus Corporation 
On June 16. 1981, the Director. NIM, on 
the recommendation of the RAC, 
approved requests from Cetus 
Corporation. 600 Bancroft Way, 
Berkeley, California 94710, for large- 
scale culture of Ekl E. coli or NVl 
Bacillus subtilis containing plasmids 
coding for human beta-1 fibroblast 
interferon. 
The request was approved with the 
understanding that Cetus Corporation 
has agreed to permit an observer, 
designated by NIM, to visit the facilities 
if NIM should choose to inspect the site. 
The principal investigators or Drs. 
Michael W. Conrad and Wolfgang M. 
Manisch. The work is to be done at the 
Pl-LS level. 
III. Johns Hopkins University 
On June 16. 1981, the Director, NIH. on 
the recommendation of the RAC, 
approved a request from Professor 
Hamilton O. Smith. Johns Hopkins 
University School of Medicine, 
Baltimore, Maryland 21205, for the large- 
scale culture of E. coli containing 
recombinant plasmids comprised of E. 
coli and Haemophilus haemolyticus 
DNA; the Haemophilus DNA consists of 
two of the genes that constitute the 
Hhall restriction/modification system. 
Professor Smith is the principal 
investigator. The work is to be done at 
the Pl-LS level. 
Dated: June 22. 1981. 
Donald S. Fredrickson, M.D., 
Director, National Institutes of Health. 
Note. — OMB's “Mandatory Information 
Requirements for Federal Assistance Program 
Announcements" (45 FR 39592) requires a 
statement concerning the official government 
programs contained in the Catalog of Federal 
Domestic Assistance. Normally NIIH lists in 
its announcements the number and title of 
affected individual programs for the guidance 
of the public. Because the quidance in this 
notice covers not only vitually every NIH 
program but also essentially every federal 
research program in which DNA recombinant 
molecule techniques could be used, it has 
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