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Federal Register / Vol. 45, No. 227 / Friday, November 21, 1980 / Notices 
in Escherichia coli K-12 and under PI 
conditions in Pseudomonas aeruginosa. 
10. Permission is granted to return to 
the host of origin Helminthosporanium 
maydis (race O) DNA which has been 
cloned in yeast strain SHY2 using the 
hybrid E. coli — yeast plasmid Ylp5. The 
cloned DNA may be returned to, and 
propagated in, Helminthosporanium 
maydis at .the P2 level of physical 
containment. 
11. Permission is granted to return 
Schizophyllum commune DNA (or yeast 
DNA) cloned in Saccharomyces 
cerevisiae with YR or 2 mu circle 
vectors to Schizophyllum commune. The 
cloned DNA may be returned to, and 
propagated in, Schizophyllum commune 
at the P2 level of physical containment. 
12. Permission is granted to return 
Wangiella dermatitidis DNA to 
Wangiella dermatitidis using an HV2 
certified Saccharomyces /E. coli hybrid 
vector. The Wangiella dermatitidis may 
be propagated at the P3 level of physical 
containment. 
13. 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 Fransico, California. Further 
development of the clones at Genentech 
has been approved under Pi + EK1 
conditions. 
14. Saccharomycopsis lipolytica may 
be used as a host for tranformation 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. 
15. 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. 
16. Permission is granted to introduce 
DNA segments from aphid transmissible 
strains into non-aphid transmissible 
strains of Cauliflower mosaic virus in 
order to study the factors determining 
aphid transmissibility. 
17. 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. 
18. Schizosaccharomyces pombe DNA 
may be cloned in Schizosaccharomyces 
pombe using approved HV1 
Saccharomyces cerevisiae/E. coli 
hybrid plasmids as vectors under Pi 
containment conditions. 
19. The pyrogenic endotoxin type A 
(Tox A) gene of Staphylococcus aureus 
may be cloned in an HV2 Bacillus 
subtilis host-vector system under P3 
containment conditions. 
20. A hybrid plasmid composed of, (1) 
E. coli plasmid pBR325, (2) the origin of 
replication and transfer genes of 
Agrobacterium tumefaciens plasmid Ti, 
(3) the thiamine gene of E. coli, and (4) 
Arabidopsis. DNA, may be transformed 
into Agrobacterium tumefaciens under 
Pi conditions. The Agrobacterium 
tumefaciens may subsequently be used 
to introduce the composite plasmid 
carrying Arabidopsis DNAand the E. 
coli thiamine gene into Arabidopsis 
plants under Pi containment conditions. 
21. Chlamydomonas reinhardi can be 
used as a host for cloning defined DNA 
segments derived from E. coli and 
Saccharomyces cerevisiae using E. coli/ 
S. cerevisiae hybrid vectors under P2 
physical containment. 
22. Candida albicans can be used as a 
host for cloning Candida albicans DNA 
following propagation of the DNA in E. 
coli K-12 or in Saccharomyces 
' cerevisiae employing an E. coli-S. 
cerevisiae hybrid plasmid vector or the 
yeast 2 micron plasmid. 
23. The Rd strain of Hemophilus 
influenzae can be used as a host for the 
propagation of the cloned Tn 10 tet R 
gene derived from E. coli K-12 
employing the non-conjugative 
Haemophilus plasmid, pRSF0885, under 
PI conditions. 
24. Zymomonas mobilis may be used 
as a host under P2 conditions for 
transformation by recombinant DNA 
derived from Pseudomonas strains that 
are non-pathogenic for animals or 
plants, and that has been cloned in an E. 
coli K-12 host. 
25. Protoplasts of Streptosporangium 
brasiliense may be transformed with a 
hybrid plasmid containing pBR322 plus a 
Streptosporangium plasmid into which 
have been incorporated specified DNA 
segments from Streptomyces species or 
an HVl approved Bacillus subtilis 
cloning vector. 
Appendix F — Certified HV2 Host-Vector 
Systems 
While the Guidelines no longer 
specify the use of E. coli K-12 EK2 or 
Saccharomyces 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, SHYl, 
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, YIp32, and 
YIp33. 
EK2 Plasmid Systems. The E. coli K- 
12 strain chi-1776. 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-1776 or 
in the sterile yeast strains, SHYl, SHY2, 
SHY 3 and SHY4: YIpl, YEp2, YEp4, 
YIp5, YEp6, YRp7, YEp20, YEp21, YEp24, 
YIp25, YIp26, YIp27, YIp28, YIp29, YIp30, 
YIp31, YIp32, YIp33. 
EK2 Bacteriophage Systems. The 
following are certified EK2 systems 
based on bacteriophage lambda: 
Vector Host 
-gt WES.-ff 
DPSOsupF 
- &WES.-B * 
DP50 supF 
-gtZ] vir.-B" 
E. coli K-12 
-gtALO.-B 
DPSOsupF 
Charon 3A 
DP50 or DP50 supF 
Charon 4A 
DP50 or DP50 supF 
Charon 16A 
DP50 or DP50sup/’ 
Charon 21A 
DP50 supF 
Charon 23A 
DF50 or DPjOsupF 
Charon 24A 
DP50 or DP50 supF 
Dated: November 14, 1980. 
Donald S. Fredrickson, 
Director, National Institutes of Health. 
OMB’8 "Mandatory Information 
Requirements for Federal Assistance Program 
Announcements” (45 FR 39592) requires a 
statement concerning the official government 
program contained in the Catalog of Federal 
Domestic Assistance. Normally NIH lists in 
its annoucements the number and title of 
affected individual programs for the guidance 
[27] 
