Federal Register / Vol. 46, No. 48 / Thursday, March, 12, 1961 / Notices 
16453 
Animals or Plants, and/or Lower 
Eukaryotes Nonpathogenic for Man, 
Animals or Plants, 
Recombinant DNA experiments 
involving prokaryotes nonpathogenic for 
man, animals, or plants, and/or lower 
eukaryotes nonpathogenic for man, 
animals or plants, and only DNA ht)m 
such sources, can be conducted under 
P3 containment (2A). Lower levels of 
physical containment may be assi^ed 
by ORDA on a case-by-case basis for 
specific donor-recipient combinations 
(see Section IV-E-l-b-{3)-{h)).” 
A new paragraph is added just before 
Section UI-O begins, as follows: 
“When the reader finds that the 
containment level given for the same 
experiment is different in two different 
sections within Part III, he may choose 
whichever of the two levels he wishes to 
use for the experiment.” 
The second paragraph (’f Section IH- 
B-3 is deleted. Revised Section ni-B-3 
now reads as follows: 
“ni-B-3. Non-HVl Systems, 
Containment levels for other classes of 
experiments involving non-HVl systems 
may be approved by the Director, NIH, 
(See Sections IV-E-l-b-(l)-(b), IV-E-1— 
b-(2Hc). and IV-E-l-b-{3Hb)).” 
Section rV-E-l-b-(2Hf) is deleted. 
A new section IV-E-l-b-{3)-{h) is 
added as follows: 
*TV-E-l-b-(3)-(h). Assigning 
containment levels for experiments hr 
which both donor and recipient are 
nonpathogenic prokaryotes and/or 
nonpathogenic lower eukaryotes (see 
Section III-0-2).” 
In order to clarify the relationship 
among Sections UI-O, UI-O-1, and lU- 
0- 2, the Unal sentence in the second 
paragraph under Part UI is amended to 
read as follows; 
"Prior BBC review is required for all 
other experiments described in the 
subsections of Part lU, including DI-O-1, 
m-O-2, etc.” 
1- B. Request for Approval of 
Scbizosaccharomyces Pombe for 
Recombinant DNA Experiments 
Dr. Benjamin D. Hall of the University 
of Washington, in letters dated 
November 3, 1980, and December 5, 
1980, requested that the fission yeast, 
Scbizosaccharomyces pombe, together 
with the S. cerevisiae/E, coli hybrid 
recombinant plasmids, be certified as an 
HVl host-vector system. Dr. Hall further 
requested that this system be included 
imder Section lU-O of the Guiddines. In 
his submission. Dr. Hall noted that 
Scbizosaccharomyces pombe is not a 
pathogen, it survives poorly outside of a 
controlled laboratory enviioiunent, it is 
rare in nature, and geneti&exchange in 
the Ascomycetes occurs only with 
closely related organisms. 
Dr. Hall’s request was published in 
the November 28, 1980 F^eral Register 
(45 FR 79387), No conunents were 
received during the thirty day comment 
period. 
The RAC, in discussing the proposal 
at the January 8-9, 1981 meeting, noted 
that specifications for HVl certification 
have been defined in Section Il-D-2-b 
of the Guidelines. It was suggested that 
only certain experiments be approved 
without certifying S. pombe as a general 
HVl host-vector system. The RAG 
therefore, recommended by a vote of 
fourteen in favor, one opposed and four 
abstentions, that DNA from non- 
pathogenic prokaryotes and lower 
eukaryotes may be cloned into 
Scbizosaccharomyces pombe species 
imder Pi containment conditions. It 
further recommended by a vote of 
fourteen in favor, none opposed and five 
abstentions, that DNA fi-om higher 
eukaryotes may be cloned in SL pombe 
species under ^ containment 
conditions. 
I accept these recommendations. Item 
18 in Appendix E of fite Guidelines is 
amended to read as follows: 
“18. DNA fiom nonpadiogenic 
prokaryotes and nonpathogenic lower 
eukaryotes may be cloned into 
Scbizosaccharomyces pombe species 
under PI cmitainment conditions. WlA 
from higher eukaryotes may be cloned in 
S, pombe species under P3 cimtainment 
conditions.” 
I-C. Request To Include Streptococcus 
Faecalis and Streptococcus Sanguis in 
Appendix A, 
Dr. Donald Clewell of die University 
of Michigan requested in letters dated 
August 13, 1980 and September 10, 1980, 
that Streptococcus faecalis be included 
along with Streptococcus sanguis in a 
sublist of Appendix A. Recombinant 
DNA molecules composed entirely of ^ 
DNA segments bom organisms within 
each sublist of Append A are exempt 
from the Guidelines under Section 1-^-4, 
which exempts "Certain specified 
recombinant DNA molecules that 
consist entirely of DNA segments from 
different species that exchange DNA by 
known physiological processes. ...” In 
support of his request. Dr. CleweU 
submitted data indicating that exchange 
of genetic information between the two 
species occurs by naturfil physiological 
processes. 
A Federd Register announcement of 
Dr. Clewell's request appeared on 
November 28, 1980 (45 1^ 79386). No 
comments were received during the 
thirty day comment period. 
The RAC discussed Dr. Clewell's 
proposal at the January 8-9, 1981 
meeting. The RAC was satisfied that the 
data demonstrated two-way plasmid 
transfer. By a vote of twelve in favor, 
two opposed and five abstentions, the 
RAC recommended that the request i>e 
approved, and that Streptococcus 
faecalis be added to sublist F of 
Appendix A. 
I accept this recommendation, and 
Appendix A, Sublist F has been 
amended to read as follows: 
"Sublist F 
1. Streptococcus sanguis 
2. Streptococcus pneumoniae 
3. Streptococcus faecalis" 
I-D. Request To Clone Saccharomyces 
Cerevisiae DNA in Tetrabymena 
Thermophilia 
Dr. Eduardo Orias of the University of 
California, Santa Barbara in a letter 
dated December 18, 1980 requested 
permission to clone Saccharomyces 
cerevisiae EWA in Tetrabymena 
thermophila using S. cerevisiae/E, coB 
hybrid plasmids. 
Dr. Orias supplied data outlining the 
genetics and ecology of Tetrabymena. 
He noted that Tetrabymena thermophila 
is a unicellular microbial eukaryote of 
no known pathogenicity. Dissemination 
of genetic information between 
members of the species by means other 
than eukaryotic conjugation has not 
been demonstrated. 
Dr. Orias* request appeared in the 
November 28, 1980 F^eral Register (45 
FR 79387). No comments were received 
during the thirty day comment period. 
During the RAC discussion of die 
proposal at the January 8-9, 1981 
meeting, it was noted ^at the proposal 
invdves nonpathogenic organisms, and 
approval at die Pi level of containment 
was recommended by a unanimous vote 
of nineteen in favor. 
1 accept this recommendation, and a 
new ent^ has been added to Appendix 
E to read as follows: 
“26. Saccharomyces cerevisiae DNA 
may be doned in Tetrahymeno 
thermophila using E. coli/S. cerevisiae 
hybrid plasmids under PI containment 
conditions.’! 
I-E. Proposal To Amend Item 4 of 
Appendix E 
Entry 4 of Appemlix E of the 
Guidelines reads as follows: 
“4. Cloned desired fragments from any 
non-jirohibited source may be 
transferred into Agrobacterium 
tumefaciens containing a Tt plasmid (or 
derivatives thereof), using a 
nonconjugative K co// plasmid vector 
[661 
