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Federal Register / Vol. 44, No. 232 / Friday, November 30, 1979 / Notices 
experiments in which there is a 
deliberate attempt to have the E. coli K- 
12 efficiently express any gene coding 
for a eukaryotic protein. 
The last two paragraphs of Section 
III— 0 are a rephrasing of what appears 
as part of Section III-A-l-b-(l) of the 
1978 Guidelines, including incorporation 
of rewording, as discussed below, in the 
first item in Part IV of this 
announcement. 
Within Section IV of the Guidelines, a 
note has been inserted in Section IV-D- 
1— c reminding the reader that “no MUA 
is required for experiments described in 
Section HI-0,” and a note has been 
inserted in Section IV-D-5-a-(l) 
reminding the reader that “no prior 
approval by the IBC is required for most 
experiments described in Section HI-0.” 
At places in the Guidelines describing 
“return to host of origin” type 
experiments (i.e., Sections IH-B-2, HI-C- 
5, HI— C— 6, HI— C— 7— a, and HI— C— 7— b), the 
phrase "appropriate containment" has 
been replaced by “Pi.” 
Section HI-A of the Guidelines, and 
its subsections, previously referring to 
experiments using E. coli K-12 as the 
host have been rewritten to refer to 
certain other certified HVl and HV2 
systems, as listed in Appendix D of the 
November 1979 Guidelines. 
It was the intent of the RAC that “the 
principle of equivalency of HV systems 
with EK systems applies at the present 
time only to the setting of containment 
levels for shotgun experiments. It does 
not apply at the present time to lowering 
of containment levels for characterized 
or purified DNA preparations and 
clones, to returning DNA segments to 
non-HVl host of origin, etc.” (Federal 
Register, July 20, 1979). Therefore, 
changes have been made in Section IH- 
A-l-a-(l), HI-A-3-a, HI-A-3-b, IV-D- 
1— c— (3), IV-D-l-e, and IV-E-l-b-(3)-(e), 
indicating that lowering of containment 
levels for characterized or purified DNA 
preparations or clones requires prior 
approval by the NIH and that IBC 
approval alone is no longer sufficient. In 
accordance with Section IV-E-l-b of 
the NIH Guidelines and based on the 
extensive analysis given above, I find 
that these proposed actions on the "E. 
coli K-12/P1 Recommendation" comply 
with the Guidelines and present no 
significant risk to health or the 
environment. 
Appendix A gives the membership at 
the Recombinant DNA Advisory 
Committee at the May 1979 meeting; 
Appendix B gives the membership at the 
September 1979 meeting. 
IV. Other Recommendations on “Major 
Actions” Made at the September 6 - 7 , 
1979 , RAC Meeting 
In addition to the "E. coli K-12/P1 
Recommendation” discussed above, five 
other recommendations on "major 
actions" were made at the September 6- 
7 RAC meeting. These are discussed 
below, and my proposed action on them 
is given. In accordance with Section IV- 
E-l-b of the NIH Guidelines, I find that 
these proposed actions comply with the 
Guidelines and present no significant 
risk to health or the environment. 
Proposed Amendment of Sections Il-D- 
l-a-(l) and III-A-l-b-(l) of the 1978 
Guidelines 
In response to a suggestion made in a 
letter of May 16, 1979, from Dr. Nickolas 
J. Panopoulos, of the University of 
California at Berkeley, proposed 
changes in the Guidelines were 
published for comment in the Federal 
Register on July 31, 1979, as follows: 
1. Proposed to be inserted at the end 
of Section II-D-l-a-(l) of the Guidelines 
were the words "except as specified 
under Section III-A-l-b-(l).” 
2. Proposed to be inserted at the end 
of section HI-A-l-b-(l) of the 
Guidelines were the words "When a 
non-conjugative vector is used, the E. 
coli K-12 host may contain conjugative 
proficient plasmids, either autonomous 
or integrated, or generalized transducing 
phages. In general, for experiments in 
this category, the E. coli K-12 host may 
contain such plasmids or phages 
provided that the physical containment 
level is raised one step.” 
During the 30-day comment period, no 
comments were received. 
At the RAC meeting on September 6- 
7, 1979, this item was discussed. A 
motion was passed by the RAC by a 
vote of 12 to 0 to: 
1. Insert at the end of Section II-D-1- 
a-(l) of the 1978 Guidelines the words 
"except as specified under Section IH- 
A-l-b-(l)”; and 
2. Insert prior to the last sentence of 
Section II-A-l-b-(l) of the 1978 
Guidelines, the words “When a non- 
conjugative vector is used, the E. coli K- 
12 host may contain conjugation 
proficient plasmids, either autonomous 
or integrated, or generalized transducing 
phages.” 
I propose to accept these 
recommendations. However, due to my 
proposed action on the “E. coli K-12/P1 
Recommendation” (as described above 
in Part HI-E of this announcement), 
further changes are necessary in 
incorporating these recommendations 
into the proposed revised NIH 
Guidelines. Thus the insert at the end of 
Section II— D— 1— a— (1 ) reads, "except as 
specified in Section IH-0,” and the 
added words "When a nonconjugative 
vector is used, the E. coli K-12 host may 
contain conjugation-proficient plasmids, 
either autonomous or integrated, or 
generalized transducing phages” appear 
in Section III— 0. 
Proposed Exemption for Pseudomonas 
Putida and Pseudomonas Fluorescens 
At its May 21-23, 1979, meeting, the 
RAC considered a request by Dr. N. 
Omston of Yale University to add 
Pseudomonas putida and Pseudomonas 
fluorscens to the exempt list in 
Appendix A of gram-negative organisms 
that exchange DNA by known 
physiological processes. The RAC, at 
that time, voted 17 to 1, with 1 
abstention, to defer action on the 
proposal, since several members felt 
that the transduction data were 
incomplete, and an error was made in 
the Federal Register notice. A request 
for additional data on the reversion 
frequencies for the transduced markers 
was made on recommending deferment. 
After receipt of additional 
documentation on the chromosomal 
genetics of Pseudomonas, the following 
notice was placed in the Federal' 
Register on July 31, 1979, for comment: 
Dr. N. Orsnston of Yale University has 
proposed, in accord with Section I— El — 4 of the 
Guidelines, that Pseudomonas putida and 
Pseudomonas fluorescens be added to the 
exempt list in Appendix A of gram-negative 
organisms that exchange DNA by known 
physiological processes. Further information 
documenting the exchange of genetic 
information between these two species and 
those in Appendix A is available from the 
Office of Recombinant DNA Activities. 
No comments were received during 
the 30-day comment period. 
At the September 6-7 RAC meeting 
the data on exchange and homology 
were discussed and a motion to add 
Pseudomonas putida and Pseudomonas 
fluorescens to the Appendix A list 
passed by a vote of 11 to 0, with 3 
abstentions. 
I propose to accept this 
recommendation and have added 
Pseudomonas putida and Pseudomonas 
fluroescens to Sublist A in Appendix A, 
of the proposed revised Guidelines. 
Cloning in Bacillus Subtilis an 
Streptomyces Coelicolor 
In response to a request from Dr. 
Stanley Cohen of Stanford University, 
the following proposal was published 
for comment in the Federal Register on 
July 31, 1979: 
(a) Bacillus subtilis strains that do not 
carry an asporogenic mutation can be used as 
hosts specifically for the cloning of DNA 
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