77372 
Federal Register / Vol. 45, No. 227 / Friday, November 21, 1980 / Notices 
DEPARTMENT OF HEALTH AND 
HUMAN SERVICES 
National Institutes of Health 
Recombinant DNA Research; Actions 
Under Guidelines 
agency: National Institutes of Health, 
PHS, HHS. 
ACTION: Notice of Actions under NIH 
Guidelines for Research Involving 
Recombinant DNA Molecules. 
SUMMARY: This notice sets forth actions 
taken by the Director, NIH, under the 
January 1980 NIH Guidelines for 
Research Involving Recombinant DNA 
Molecules (45 FR 6724). 
EFFECTIVE DATE: November 21, 1981. 
FOR FURTHER INFORMATION CONTACT. 
Additional information can be obtained 
from Dr. William J. Gartland, Office of 
Recombinant DNA Activities (ORDA), 
National Institutes of Health, Bethesda, 
Maryland 20205. (301) 496-6051. 
SUPPLEMENTARY INFORMATION: I am 
promulgating today several actions 
under the NIH Guidelines for Research 
Involving Recombinant DNA Molecules. 
These were reviewed and recommended 
for approval by the Recombinant DNA 
Advisory Committee (RAC) at its 
meeting on September 25-26, 1980. In 
accordance with Section IV-E-l-b of 
the NIH Guidelines, I find that these 
actions comply with the Guidelines and 
present no significant risk to health or 
the environment. 
This announcement provides 
background information on the actions 
including an analysis of correspondence 
concerning these actions received up to 
Octoiber 31, 1980. 
Immediately following this 
announcement, there appears in the 
Federal Register a copy of revised NIH 
Guidelines for Research Involving 
Recombinant DNA Molecules. These 
revised Guidelines differ from the 
previous version of the Guidelines 
promulgated on January 29, 1980 (45 FR 
6724) by incorporating within them: 
changes in the Guidelines which were 
recommended at the RAC meeting of 
March 6-7, 1980, and promulgated on 
April 14, 1980 (45 FR 25366): changes in 
the Guidelines which were 
recommended at the RAC meeting of 
June 5-6, 1980, and promulgated on July 
29, 1980 (45 FR 50524); and changes in 
the Guidelines which were 
recommended at the RAC meeting of 
September 25-26, 1980 and which are 
discussed in this announcement. 
I. Proposal To Introduce Genes Cloned 
in E. Coli K-12 into Arabidopsis Plants 
Through the Use of Agrobacterium 
Tumefaciens Carrying an E. Coli/Ti 
Hybrid Plasmid Vector 
In a letter dated March 21, 1980, Dr. 
Donald J. Merlo of the University of 
Missouri-Columbia, requested an 
evaluation of the containment 
appropriate for the following protocol:' 
A. A hybrid vector, constructed from 
the E. coli plasmid pBR325 and the 
origin of replication and transfer genes 
of Agrobacterium tumefaciens plasmid 
Ti, will be cloned in E. coli K-12. 
B. Arabidopsis DNA will be 
introduced into the E. coli I Ti hybrid 
plasmid and clone in E. coli K-12. 
C. The thiamine gene of E. coli will 
bed introduced into the E. coli I Ti vector 
carrying Arabidopsis DNA and cloned 
in E. coli K-12. 
D. The hybrid plasmid into which 
Arabidopsis DNA and the thiamine gene 
have been ligated will be transformed 
into Agrobacterium tumefaciens. 
E. Agrobacterium tumefaciens will be 
used to introduce the E. coli/ Ti plasmid 
vector carrying the E. coli Thiamine 
gene and Arabidopsis DNA into 
Arabidopsis plants. 
The RAC discussed Dr. Merlo's 
proposed protocol at the June 5-6, 1980 
meeting. The RAC stated that the first 
three steps of the proposed experiment 
are covered by Section III-O of the 
Guidelines. It was agreed that steps D 
and E are covered by Appendix E and 
that P3 containment is indicated. 
However, as the proposed inserted 
sequences were well characterized, and 
as Arabidopsis DNA will be returned to 
Arabidopsis, the RAC suggested that 
containment for steps D and E could be 
lowered to P2. 
Since this had not been published in 
the Federal Register for comment prior 
to the June 5-6, 1980, RAC meeting, this 
proposed lowering would, however, 
have to be published in the Federal 
Register for thirty days of comment and 
re-considered at the September 25-26, 
1980 meeting. 
In a letter dated July 22, 1980, Dr. 
Merlo described the proposed 
experiments in greater detail and 
requested that the containment 
conditions for steps D and E be lowered 
at least to P2 and preferably to PL 
Dr. Merlo’s proposal was published in 
the August 21, 1980, Federal Register (45 
FR 55924). No comments were received 
during the thirty day comment period. 
The RAC, at its September 25-26, 1980 
meeting, once again discussed this issue. 
A motion to approve the experiments at 
the P2 level of containment failed by a 
vote of 4 in favor, 9 opposed, with 3 
abstentions. Then, by a vote of 12 in 
favor, 2 opposed, with 3 abstentions, the 
RAC recommended approval of the 
proposed experiments under PI 
containment. 
I accept this recommendation, and 
text has been added to Appendix E of 
the Guidelines indicating this. 
II. Request for Certification of a Bacillus 
Stearothermophilus Derived Plasmid as 
the Vector Component of an HV1 
Bacillus Subtilis Host-Vector System 
Dr. David B. Wilson of Cornell 
University, in a letter dated May 12, 
1980, requested certification of a 
Bacillus stearothermophilus derived 
plasmid, pABl24, as the vector 
component of an HV1 Bacillus subtilis 
host-vector system. The HVl-certified 
Bacillus subtilis strain RUB331 would be 
used as the host component. 
The RAC discussed this proposal at 
the June 5-6, 1980 meeting. It was noted 
that Bacillus stearothermophilus is a 
non-pathogenic thermophile found in 
soil near hot springs, on compost, etc. In 
addition, plasmid pABl24 can be 
transformed into and maintained in 
Bacillus subtilis by natural 
physiological methods; pABl24 might, 
therefore, be considered a plasmid 
indigenous to Bacillus subtilis. The 
committee agreed that this proposal 
should be published in the Federal 
Register for comment an^acted upon at 
the September 25-26, 1980 meeting. 
Dr. Wilson’s proposal was published 
for thirty days of comment in the August 
21, 1980, Federal Register (45 FR 55925). 
No comments were received during this 
period. The RAC once again discussed 
the proposal at the September 25-26, 
1980 meeting. The RAC recommended 
approval of the proposal by a vote of 18 
in favor, none opposed. 
I accept this recommendation, and 
text has been added to Appendix D of 
the Guidelines indicating this. 
III. Request for Certification of a 
Bacillus Subtilis Strain as the Host 
Component of an HV2 Host-Vector 
System 
On March 28, 1980, Dr. William Burke, 
Jr., of Arizona State University, 
requested certification of Bacillus 
subtilis strain ASB298 as the host 
component of an HV2 host-vector 
system. Dr. Burke’s request appeared in 
the Federal Register of April 30, 1980 (45 
FR 28907). No comments were received 
during this period. Additional 
information was submitted by Dr. Burke 
in a letter dated May 13, 1980. 
Dr. Burke's proposal was evaluated by 
an ad hoc working group and 
subsequently presented to the RAC at 
the June 5-6, 1930 meeting. One RAC 
[206] 
