16 
be used for cloning CNA from any nonprohibited sources, using 
vectors indigenous to ^ subtil is , under the same conditions 
specified by RAC for ^ ooli K-l2 emd ^ oerevisiae host-vector 
systems. 
(2) Bacillus subtilis strains that do not carry an asporogenous 
mutation can be used with vectors indigenous to B. subtilis 
for the cloning of CfA fran amy CDC Cleiss 1 organism under 
P2 conditions. 
(3) Bacillus subtilis strains that do not carry eisporogenic 
mutation can be used with vectors indigenous to ^ subtilis 
mder PI conditions for the cloning of CNA from amy Class 1 
Bacillus species. 
Dr. williems said that Bacillus subtilis is probably the most extensively 
understood gram-positive o^g^lnl9m, both genetically and biochemicedly. It 
is capable of both generalized and speciedized transduction and has been 
widely used in the industrial sector in the prodxxrtion of em array of 
antibiotics. It may be particularly well suited for certain types of 
recorab inant DNA experiments, as Bacillus strains have the capacity to 
secrete a variety of proteins. Bacillus subtilis is nonpathogenic and 
is not kno%ai to exchange genetic information with pathogens. 
Dr. Williams directed the ocmmittee's attention to data comparing the 
survivability of B. subtilis and E. roli K-12 in soil or water samples; 
E. ooli survives Better than B. aiBtilis over a five day period. Additional 
3a ta demonstrate that B. subtTTis spores placed in a rumnalian intestine 
reurely speculate, and IT they do the vegetative cells quickly die. 
Dr. Williams cecormended approval of the request. Dr. Holmes seconded the 
motion. 
Dr. Qottestwi, noting that currently certified HVl ^ subtilis host-vector 
systems only employ certain specified plasmids, pointed out that Dr. Dean's 
proposal would also permit use of phage vectors. She requested additional 
infonnation concerning the proposed phage vectors. Dr. Dean replied that 
the host ranges of the Bacillus phages are very narrow. In his experience, 
transformation aif fords greater possibilities euid avenues of genetic exchange. 
Dr. Goldstein asked if ^ subtilis engineered to excrete recombinant 
proteins is a concern. Dr. Novick replied that ^ subtilis does not colon- 
ize the manmalian gastrointestin^d tract. He personally did not regard 
the excretion of cloned proteins into soil as potentially hazardous. 
Dr. Talbot requested clarification of Dr. Williams' motion. He asked if 
the intent was that the asporogenic ^ subtilis strains would be exempted 
frem the Guidelines as had been recommended by the RAC for ^ ooli K-12 
and S. ^revisiae eeurlier in the meeting. Dr. Williams replied that he 
interSed they would. Noting that the language of the proposal would permit 
the use of any "indigenous vector," Dr. Talbot questioned whether the 
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