MINUTES OF MEETING - June 5-6 
17 
conjugation proficient Escherichia coli K-12, this section could only 
cover Ff phages grown on hosts which are carrying a transfer-defective 
plasmid. He said the Phage Working Group met by conference call and, 
after evaluating the issues, agreed unanimously that Ff phages be inclu- 
ded under Section III-O. He then moved approval of the language published 
in the Federal Register of April 30, 1980. 
Dr. Goldstein requested further discussion of the ecology and host-range 
of the filamentous phages. Dr. Gottesman said that during the conference 
call, two points were made: (1) the Ff host-range is not as extended as 
some thought, and (2) the filamentous phages do not stably maintain infor- 
mation cloned into the vector. Dr. Campbell pointed out that these parti- 
cular male-specific phages are rarely isolated in nature. He said the 
RNA phages appear to be more abundant, while the DNA phages are rare. 
Dr. Setlow called the question. The RAC recommended acceptance, by a 
vote of fourteen in favor and none apposed, of the language in the Federal 
Register including Ff phages under Section III-O. 
VIII. PROPOSED HV2 BACILLUS SUBTILIS HOST-VECTOR SYSTEMS 
Dr. Gottesman introduced the request (tab 875, 881/11, 899) from 
Dr. William F. Burke of Arizona State University, for HV2 certification 
of Bacillus subtilis strain ASB298. She said that Bacillus subtilis 
strain ASB298 contains a variety of mutations. Two sets of mutations 
block sporulation. A deletion mutation leads to cell lysis under certain 
growth conditions. Another mutation prevents cell growth in the absence 
of the rare amino acid, D-alanine. Another mutation confers streptomycin- 
dependence . 
Dr. Gottesman said that an ad hoc group discussed this request by confer- 
ence call. She said that overall the group was satisfied with the data 
presented by Dr. Burke. She said the only issue raised concerning a 
Bacillus subtilis HV2 system was the question of transfer of genetic 
information from this strain to other Bacillus by transformation. She 
noted that no mutation Dr. Burke has introduced into this strain blocks 
transfer out of genetic information through transformation; the only 
constraint in this Bacillus subtilis system is that ASB298 may not grew 
very well. Dr. Gottesman added that in Bacillus subtilis , plasmid vec- 
tors do not transfer by conjugation. She noted that recombinant material 
could, however, be transferred by phage vectors and suggested that a 
Bacillus subtilis HV2 be restricted to plasmid vectors. 
Dr. Young said that competent Bacillus subtilis cells can excrete DNA. 
He felt that any Bacillus subtilis strain excreting DNA is analogous to 
a conjugation proficient Escherichia coli . Dr. Gottesman said the ad 
hoc group had discussed the possibility of requesting that Dr. Burke 
evaluate transformation frequencies in a "flower pot" protocol. 
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