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Federal Register / Vol. 44, No. 232 / Friday, November 30, 1979 / Notices 
References 
1. Classification of Etioiogic Agents on the 
Basis of Hazard. (4th Edition, July 1974). U.S. 
Department of Health, Education, and 
Welfare, Public Health Service, Center for 
Disease Control, Office of Biosafety, Atlanta, 
Georgia 30333. 
2. National Cancer Institute Safety 
Standards for Research Involving Oncogenic 
Viruses (October 1974). U.S. Department of 
Health, Education, and Welfare Publication 
No. (NIH) 75-790. 
3. U.S. Department of Agriculture, Animal 
and Plant Health Inspection Service. 
Appendix C 
Section I-E-5 states that exempt from these 
Guidelines are "Other classes of recombinant 
DNA molecules, if the Director, NIH, with 
advice of the Recombinant DNA Advisory 
Committee, after appropriate notice and 
opportunity for public comment, finds that 
they do not present a significant risk to 
health or the environment. (See Section FV- 
EI— 1— b — (1) — (d).) Certain classes are exempt as 
of publication of these Revised Guidelines. 
The list is in Appendix C." 
Under exemption I-E-5 of these Revised 
Guidelines are those recombinant DNA 
molecules that are propagated and 
maintained in cells in tissue culture and that 
are derived entirely from non-viral 
components (that is, no component is derived 
from a eukaryotic virus). 
Appendix D 
As noted above at the beginning of Section 
III— A, certain HVl and HV2 host-vector 
systems are assigned containment levels as 
specified in the subsections of Section III-A. 
Those so classified as of publication of these 
Revised Guidelines are listed below. 
* HVl — Unmodified laboratory strains of 
Saccharomyces cerevisiae 
* HVl — The following specified strains of 
Neurospora crassa which have been modified 
to prevent aerial dispersion: (1) ini 
(inositolless) strains 37102, 37401, 46316, 
64001 and 89601. 
(2) csp-1 strain UCLA37 and csp-2 strains 
FS 590, UCLA101 (these are conidial 
separation mutants). 
(3) eas strain UCLA191 (an "easily 
wettable" mutant. 
HVl — Asporogenic mutant derivatives of 
B. subtilis. These derivatives must not revert 
to sporeformers with a frequency greater than 
10" data confirming this requirement must 
be presented to NIH for certification. The 
following plasmids are accepted as the vector 
components of certified B. subtilis HVl 
systems: pUBllo, pCl94, pSl94, pSA2100, 
pEl94, pT127, pUBll2, pC221, pC223. 
* HV2 — The following sterile strains of 
Saccharomyces cerevisiae, all of which have 
the ste-VC9 mutation, SHY1, SHY2, SHY3, 
and SHY4. The following plasmids are 
certified for use: YIpl, YEp2, YEp4, YIp5, 
YEp6, YRp7, YEp20, YEp21, YEp24, YIp25, 
YIp26, YIp27, YIp28, YIp29, YIp30, YIp31, 
■ These follow the assigned containment levels as 
specified in the subsections of Section III-A with 
one exception. This exception is that experiments 
involving complete genomes of eukaryotic viruses 
will require P3 + HV1 or P2 + HV2 rather than the 
levels given in the subsections of Section III-A 
Yip32, and YIp33. These plasmids can be 
considered EK2 vectors when propagated in 
1776. 
Appendix E 
As noted in the subsections of Section IV- 
E-l-b-(l) the Director, NIH, may take certain 
actions with regard to the Guidelines after 
public notice and RAC consideration. 
Some of the actions taken to date include 
the following: 
• The following experiment has been 
approved. The cloning in B. subtilis, under P2 
conditions, of DNA derived from 
Saccharomyces cerevisiae using EK2 plasmid 
vectors provided that an HVl B. subtilis host 
is used. 
• Unmodified laboratory strains of 
Neurospora crassa can be used in all 
experiments for which HVl N. crassa 
systems are approved provided that these are 
carried out at physical containment one level 
higher than required for HVl. However, if P3 
containment is specified for HVl N. crassa, 
this level is considered adequate for 
unmodified N. crassa. For P2 physical 
containment, special care must be exercised 
to prevent aerial dispersal of macroconidia, 
including the use of a biological safety 
cabinet. 
• P2 physical containment shall be used for 
DNA recombinants produced between 
members of the genera Streptomyces and 
Micromonospora except for those species 
which are known to be pathogenic for man, 
animals or plants [2AJ. 
• Cloned desired fragments from any non- 
prohibited source may be transferred into 
Agrobacterium tumefaciens containing a Ti- 
plasmid (or derivatives thereof), using a 
nonconjugative E. coli plasmid vector 
coupled to a fragment of the Ti plasmid and / 
or the origin of replication of an 
Agrobacterium plasmid, under containment 
conditions one step higher than would be 
required for the desired DNA in HVl systems 
(i.e. one step higher physical containment 
than that specified in the subsections of 
Section III-A). Transfer into plant parts or 
cells in culture would be permitted at the 
same containment level (one step higher). 
• Bacillus subtilis strains that do not carry 
an asporogenic mutation can be used as hosts 
specifically for the cloning of DNA derived 
from E. coli K-12 and Streptomyces 
coelicolor using NIH-approved 
Staphylococcus aureus plasmids as vectors 
under P2 conditions. 
• Streptomyces coelicolor can be used as a 
host for the cloning of DNA derived from B. 
subtilis, E. coli K-12, or from S. aureus 
vectors that have been approved for use in B. 
subtilis under P2 conditions. 
Dated: November 26, 1979. 
Donald S. Fredrickson, 
Director, National Institutes of Health. 
[FR Doc. 79-38841 Filed 11-29-79: 8:45 am) 
BILLING CODE 4110-08-M 
Recombinant DNA Research; 
Proposed Actions Under Guidelines 
AGENCY: National Institutes of Health, 
PHS, HEW. 
ACTION: Notice of proposed actions 
under NIH Guidelines for Research 
Involving Recombinant DNA Molecules. 
SUMMARY: This notice sets forth actions 
proposed by the Director, NIH, under 
the 1978 NIH Guidelines for Research 
Involving Recombinant DNA Molecules 
(43 FR 60108), and introduces the 
publication of proposed NIH Guidelines. 
FOR FURTHER INFORMATION CONTACT: 
Additional information can be obtained 
from Dr. William J. Gartland, Office of 
Recombinant DNA Activities, National 
Institutes of Health, Bethesda, Maryland 
20205. (301) 496-6051. 
SUPPLEMENTARY INFORMATION: I am 
today issuing for public comment 
proposed revised NIH Guidelines for 
Research Involving Recombinant DNA 
Molecules. This action is taken in 
accordance with Section IV-E-l-b(l) of 
the NIH Guidelines (43 FR 60126) which 
says, “The Director’s proposed decision, 
at his discretion, may be published in 
the Federal Register for 30 days of 
comment before final action is taken." 
This announcement is both a "Decision 
Document" explaining the background 
and reasons for the proposed decision 
and an Environmental Impact 
Assessment. Immediately following this 
announcement there appears a copy of 
the proposed revised NIH Guidelines. 
Both the Decision Document/ 
Environmental Impact Assessment and 
the proposed revised Guidelines are 
issued for public comment for a period 
of 30 days. Written comments and 
inquiries should be addressed to the 
Director, National Institutes of Health, 
Bethesda, Md. 20014. All comments 
received will be available for public 
inspection at the Director's office on 
weekdays (Federal holidays excepted) 
between the hours of 8:30 a.m. and 5 
p.m. The structure of this Decision 
Document/Environmental Impact 
Assessment is as follows: 
I. History of the NIH Guidelines Through 
1978. 
II. Revision of the December 1978 
Guidelines. 
III. The "E. coli K-12/P1 Recommendation " 
Made by the RAC at the September 6-7, 1979, 
Meeting. 
IV. Other Recommendations Made on 
“Major Actions " by the RA C at the 
September 6-7, 1979, Meeting. 
I. History of the NIH Guidelines Through 
1978 
The history leading to the issuance of 
original 1976 NIH Guidelines for 
Recombinant DNA Research is 
described in detail in the Environmental 
Impact Statement on the 1976 
Guidelines, and in the "Decision 
Document" accompanying the 
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