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Federal Register / Vol. 45, No. 73 / Monday, April 14, 1980 / Notices 
DEPARTMENT OF HEALTH, 
EDUCATION, AND WELFARE 
National Institutes of Health 
Recombinant DNA Research; Actions 
Under Guidelines 
AGENCY: National Institutes of Health, 
PHS, HEW. 
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 
1980 NIH Guidelines for Research 
Involving Recombinant DNA Molecules 
(45 FR 6724). 
EFFECTIVE DATE: April 14, 1980. 
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 major 
actions under the NIH Guidelines for 
Research Involving Recombinant DNA 
Molecules. These proposed actions were 
published for comment in the Federal 
Register of January 31, 1980, and 
reviewed and recommended for 
approval by the Recombinant DNA 
Advisory Committee (RAC) at its 
meeting on March 6-7, 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. 
Part I of this announcement provides 
background information on the actions. 
Part II provides a summary of the major 
actions. 
I. Decisions on Actions Under 
Guidelines 
I-A. Amendment of Section III-O of the 
Guidelines 
In response to a letter of December 21, 
1979, from Dr. Stuart Levy of Tufts 
University the following notice appeared 
in the Federal Register of January 31, 
1980: 
“Dr. Stuart Levy of Trufts University 
School of Medicine has'recommended an 
amendment to Section III-O of the Proposed 
Revised Guidelines (44 FR 69210). Dr. Levy 
notes that the plasmid studied in his risk- 
assessment studies was a nonconjugative 
poorly mobilizable plasmid. He suggests that 
a distinction be made between 
nonconjugative plasmids and nonconjugative 
plasmids which- are also poorly mobilizable; 
such as amendment would provide additional 
biological containment. The relevant portion 
of Section III-O of the Proposed Revised 
Guidelines which would be amended reads 
as follows: 
* * * Other experiments using E. coli K-12 
shall use PI physical containment and, except 
as specified in the last paragraph of this 
section, an EKl host-vector system (i.e., (a) 
the host shall not contain conjugation- 
proficient plasmids or generalized 
transducing phages, and (b) lambda or 
lambdoid bacteriophages or nonconjugative 
plasmids shall be used as vectors)* * *. The 
amended section proposed by Dr. Levy would 
read as follows (amended text in italic): 
* * * Other experiments using E. coli K-12 
shall use PI physical containment and, 
except as specified in the last paragraph of 
this section, an EKl host-vector system (i.e., 
(a) the host shall not contain conjugation- 
proficient plasmids or generalized 
transducing phages, and (b) lambda or 
lambdoid bacteriophages or nonconjugative 
poorly mobilizable (able to be mobilized at a 
frequency of less than Iff 5 by a derepressed 
conjugative plasmid in the same cell) 
plasmids shall be used as vectors )' * *.” 
No comments were received during 
the thirty day comment period. 
This proposed amendment to the 
language of Section III-O was 
considered by the RAC at its March 6-7, 
1980 meeting. In the discussion it was 
pointed out that insertion into the 
Guidelines of the requirement for a 
"poorly mobilizable” plasmid would 
then necessitate specification of what 
tests, under which conditions, must be 
performed to show that a particular 
plasmid was or was not “poorly 
mobilizable." Instead the RAC 
recommended by a vote of eleven in 
favor, none opposed and five 
abstentions, that reference to a new 
footnote be added to Section III-O, the 
footnote to read as follows: 
“A subset of non-conjugative plasmid 
vectors are also poorly mobilizable (e.g., 
pBR322, pBR313). Where practical, these 
vectors should be employed.” 
I accept this recommendation. 
I-B. Proposed Amendment of Section 
III-O of the Guidelines 
In response to suggestion made in a 
letter of December 27, 1979, from Dr. 
Kent Wilcox of the Medical College of 
Wisconsin, a notice proposing to amend 
the language of Section III-O was 
published in the Federal Register of 
January 31, 1980. The relevant part of 
Section III-O currently reads as follows: 
"* * * An exception, however, which does 
require prior review and approval by the IBC 
is any experiment in which there is a 
deliberate attempt to have the E. coli K-12 
efficiently express any gene coding for a 
eukaryotic protein.* * *” 
Dr. Wilcox suggests that for clarity, 
this sentence be modified to read as 
follows: 
“* * * An exception, however, which does 
require prior review and approval by the IBC 
is any experiment in which there is a 
deliberate attempt to have the E. coli JK-12 
efficiently express as a protein product the 
information carried in any gene derived 
either from a eukaryotic organism or from 
any virus or viroid which infects a eukaryotic 
organism.* * *" 
During the thirty day comment period, 
no comments were received. 
At the RAC meeting on March 6-7, 
1980, it was agreed that the proposed 
new wording clarifies the intent of this 
sentence. A motion accepting this 
amendment to the language of Section 
III-O was passed by the RAC by a vote 
of eighteen in favor, none opposed and 
no abstentions. 
I accept this recommendation. 
I-C. Request to Include all Species of 
the Genus Erwinia in Appendix A, 
Sublist A 
In response to a letter of December 27, 
1979, the following notice appeared in 
the Federal Register of January 31, 1980: 
"In response to the proposed revised 
Guidelines published in the Federal Register 
on November 30, 1979 (44 FR 69210), Dr. 
Clarence Kado of the University of 
California, Davis, has requested that the NIH 
consider the inclusion of all Erwinia species 
in Sublist A Appendix A. Currently only one 
Erwinia species, E. amylovora, is included on 
the list. Dr. Kado suggests that consideration 
be given to all Erwinia species on the basis of 
an exchange of genetic material by natural 
means with E. coli and a 20% DNA-DNA 
homology of E. coli with Erwinia species.” 
During the thirty day comment period 
no comments were received. 
At the RAC meeting of March 6-7, 1960, the 
data submitted by Dr. Kado were reviewed. 
By a vote of fourteen in favor, none opposed 
and three abstentions the RAC accepted a 
motion to change the entry in Appendix A, 
Sublist A of the Guidelines from "Erwinia 
amylovora" to "Genus Erwinia." 
I accept this recommendation. 
I-D. Request for Approval To Clone 
Exo toxin A Gene of Pseudomonas 
Aeruginosa 
In a letter of November 19, 1979, Dr. C. 
W. Shuster of Case Western Reserve 
University requested that the RAC 
consider the status under the Guidelines 
of cloning the exotoxin A gene of 
Pseudomonas aeruginosa in Escherichia 
coli. At its December 6-7, 1979 meeting, 
the RAC indicated that Dr. Shuster's 
request appeared to require an 
exception to a prohibition. Accordingly, 
the following notice was published in 
the January 31, 1980 Federal Register for, 
a thirty day comment period: 
"Dr. C. W. Shuster of Case Western 
Reserve University has requested the RAC to 
consider whether the cloning of the exotoxin 
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