Federal Register / Vol. 45, No. 20 / Tuesday, January 29, 1980 / Notices 
6749 
certified B. subtilis HV1 systems: 
pUBllo. pCl94. pSl94. pSA2100. pEl94, 
pT127. pl!Bll2. 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: Ylpl, YEp2. YEp4. 
YIp5, YEp6. YRp7. YEp20. YEp21. YEp24. 
Ylp25. YIp26. Ylp27. Ylp28. Ylp29. YIp30. 
Ylp31. YIp32. and YIp33. These plasmids 
can be considered EK2 vectors when 
propagated in chi-1776. 
Appendix E 
As noted in the subsections of Section IV- 
E— 1— b— (1 ) the Director. NTH. 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 (2A). 
• Cloned desired fragments from any non- 
prohibited source may be tranferred 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. 
• Certain cloned segments of Anabena 
DNA may be transferred into Klebsiella 
under P2 physical containment. 
• Permission is granted to clone foot-and- 
mouth disease virus in the EKlCV host-vector 
system consisting of E. coli K-12 and the 
vector pBR322. all work to be done at the 
Plum Island Animal Disease Center. 
Dated: January 23. 1980. 
Donald S. Fredrickson. 
Director. National Institutes of Health 
|FR Doc. 80-2822 Filed 1 - 28 - 80 : 0 45 *m| 
BILUNG CODE 4110-08-M 
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