Federal Register / Vol. 45, No. 147 / Tuesday, July 29, 1980 / Notices 
50531 
for Smallpox Research. Center for Disease 
Control, in Atlanta). 
Each mention of Alastrim, Smallpox 
and Whitepox in Appendix B should be 
followed by the symbol * * *. 
II- U. Amendment of Appendix D 
Appendix B is amended to read: 
As noted above at the beginning of Section 
III— A. certain HV1 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* — The following specified strains of 
Neurospora crassa which have been modified 
to prevent aerial dispersion: 
(1) ini (inositolless) strains 37102, 37401, 
46318, 64001 and 89601. 
(2) csp-l strain UCLA37 and csp-2 strains 
FS 590, UCLA101 (these are conldial 
separation mutants). 
(3) eas strain UCLA191 (an “easily 
wettable” mutant). 
HVl — Asporogenic mutant derivatives B. 
subtilis. The B, subtilis HVl 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, pE194, pT127, 
pUBll2, pC221, pC223. B. subtilis strain RUB 
331 has been certified as the host component 
of HVl systems based on these plasmids. 
Il-V. Amendments of Appendix E 
The following sections are added to 
Appendix E: 
• Certain specified clones derived from 
segments of the Foot-and-Mouth 
Disease Virus may be transferred 
from Plum Island Animal Disease 
Center to the facilities of Genentech, 
Inc., of South San Francisco, 
California. Further development of the 
clones at Genentech has been 
approved under Pi + EKl conditions. 
• Sacchromycopsis lipolytica may be 
used as a host for transformation with 
defined Escherichia coli/ 
Saccharomyces cerevisiae hybrid 
plasmids and the hybrid plasmids 
may be used for cloning S. lipolytica 
DNA in E. coli and returning the 
cloned DNA to S. lipolytica. 
• Conjugative plasmids or transducing 
phages may be employed in 
recombinant DNA experiments when 
employing E. coli as host when a 
small defined segment of Adenovirus 
2 DNA is employed as linker DNA. 
• Permission is granted to introduce 
DNA segments from aphid 
'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 -t- HVl or P2 + HV2 rather than the 
levels given in the subsections of Section III— A. 
transmissible strains into non-aphid 
transmissible strains of Cauliflower 
mosaic virus in order to study the 
factors determining aphid 
transmissibility. 
• Permission is granted to return Mucor 
racemosus DNA which has been 
cloned in Saccharomyces cerevisiae 
host-vector systems to Mucor 
racemosus. In addition, permission is 
granted to transform Mucor 
racemosus with S. cerevisiae vectors 
with or without cloned S. cerevisiae 
sequences. These manipulations may 
be performed under P2 conditions. 
• Many experiments utilizing E. coli or 
S. cerevisiae host-vector systems are 
now covered by Section III— O of the 
Guidelines. This section states, in 
part, that "* * * experiments using E. 
coli K-12 or laboratory strains of S. 
cerevisiae shall use PI physical 
containment and, except as specified 
in the' last paragraph of this section, 
an HVl host/ vector system * * *" 
While the Guidelines no longer 
specifiy the use of EK2 or S. cerevisiae 
HV2 systems, investigators may wish to 
employ these systems in specific 
instances. The currently certified EK2 
and HV2 systems are: 
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, YEp0, YRp7, YEp20, . 
YEp21, YEp24, YIp25, YIp20, YIp27, 
YIp27, YIp28, YIp29, YIp30, YIp31, YIp32 
and YIp33. 
EK2 Plasmid Systems. The E. coli K- 
12 strain chi-1770. The following 
plasmids are certified for use: pSClOl, 
pMB9, pBR313, pBR322, pDH24, pBR327. 
The following E. coli/S. cerevisiae 
hybrid plasmids are certified as EK2 
vectors when used in E. coli chi-1770 or 
in the sterile yeast strains, SHYl, SHY2, 
SHY3 and SHY4: YIpl, YEp2, YEp4, 
YIp5, YEp0, YRp7, YEp20, YEp21, YEp24, 
YIp25, YIp20, YIp27, YIp28, YIp29, YIp30, 
YIp31, YIp32, YIp33. 
EK2 Bacteriophage Systems. The 
following are certified EK2 Systems 
based on bacteriophage lambda: 
Vector 
X gt WES. X B' 
X gt WES. X B' 
X gtr Zlvin. X B' 
gtALO. X B 
Charon 3A 
Charon 4A 
Charon 16A 
Charon 21 A 
Charon 23A 
Charon 24A 
Host 
DP50supF 
DP50supF 
E. coli K— 12 
DP50supF 
DP50 or DP50supF 
DP50 or DP5QsupF 
DP50 or DP50supF 
DPSOsupF 
DP50 or DP50supF 
DP50 or DP50supF 
Additional Announcements of the 
Director, NIH 
In addition, two further changes are 
being promulgated in order to clarify the 
intent of the language in two sections of 
the Guidelines. 
I. Addition of Subheading to Section I-D 
A new subheading; “I-D (1-0)," is to 
be added to the penultimate paragraph 
in Section I-D to emphasize that this 
paragraph applies to I-D-l through I-D- 
0. The amended paragraph reads as 
follows: 
I-D (1-6). Experiments in Categories I-D-l 
to I-D-6 may be excepted (4) from the 
prohibitions (and will at that time be 
assigned appropriate levels of physical and 
biological containment) provided that these 
experiments are expressly approved by the 
Director, NIH, with advice of the 
Recombinant DNA Advisory Committee after 
appropriate notice and opportunity for public 
comment (See Section IV-E-l-b-(l)-(e).) 
II. Amendment of Appendix E, Item 6 
Item 8 in Appendix E is amended to 
read as follows: 
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, using as a vector 
any plasmid indigenous to Streptomyces 
coelicolor or able to replicate in that host by 
natural biological mechanisms. 
Dated: July 18. 1980. 
Donald S. Fredrickson, 
Director, National Institutes of Health. 
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 NIH lists in 
its announcements the number and title of - 
affected individual programs for the guidance 
of the public. Because the guidance in this 
notice covers not only virtually every NIH 
program but also essentially every federal 
research program in which DNA recombinant 
molecule techniques could be used, it has 
been determined to be not cost effective or in 
the public interest to attempt to list these 
programs. Such a list would likely require 
several additional pages. In addition, NIH 
could not be certain that every federal 
program would be included as many federal 
agencies, as well as private organizations, 
both national and international, have elected 
to follow the NIH Guidelines. In lieu of the 
individual program listing, NIH invites 
readers to direct questions to the information 
address above about whether individual 
programs listed in the Catalog of Federal 
Domestic Assistance are affected. 
NIH programs are not covered by OMB 
Circular A-95 because they fit the description 
of “programs not considered appropriate" in i 
Section 8(b) (4) and (5) of that Circular. 
i 
i 
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[FR Doc. 80-22365 Filed 7-28-80; 8:45 am) 
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