SEPTEMBER 6-7 - MINUTES OF MEETING 
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XVI. BACILLUS SUBTILIS HV1 SYSTEMS 
Dr. Broadbent presented a report on the B. subtilis nonsporulating strain 
RUB 331 (736) as submitted by Dr. Gary Wilson of the University of Rochester. 
He reported that the mutant reverts to spore formation with a frequency 
of less than 10 ” 7 . It lyses in complex medium and revertants are thus 
readily identified. It is more susceptible to drying than the parent 
strain. Under conditions required for sporulation of the parent strain, 
90% of RUB 331 become nonviable. The strain is well-characterized and 
meets the requirements for HVl certification. Dr. Broadbent moved to 
certify this strain as the host component of a B. subtilis HVl certified 
system. The RAC accepted this motion by a vote of twelve for, none oppose, 
with five abstentions. Dr. Young disqualified himself. 
XVII. PROTOCOLS FOR ASSIGNMENT OF CONTAINMENT LEVELS 
A. Cloning in Bacillus Subtilis and Streptcmyces Coelicolor 
Dr. Broadbent described the proposals submitted by Dr. Stanley Cohen 
of Stanford University (707/4, 722) requesting approval to perform at the 
P2 level of containment experiments a) using sporulating B. subtilis strains 
as hosts for the cloning of DNA derived from E. coli K-12 and S. coelicolor 
using NIH approved Staphylococcus aureus plasmids as vectors and b) using 
Streptomyces coelicolor as a host for the cloning of DNA derived frcm B. 
subtilis , E. coli K-12 or frcm S. aureus vectors that have been approved 
for use in B. subtilis . Dr. Broadbent chose to deal first with section 
b) of this proposal. Dr. Broadbent noted that Streptomyces spores are not 
particularly resistant to heat, i.e., autoclaving will kill the spores. 
The spores do not germinate readily. Dr. Broadbent recommended approval 
of part b) of this proposal. The motion was called with the result that 
three members supported the motion, two opposed, with eight abstentions. 
Mr. Thornton noted that Dr. Cohen is asking the RAC to reverse decisions 
made at the last two meetings. Mr. Thornton added that he felt the burden 
of reversing a decision was great and there was not sufficient new information 
to justify voting for this proposal. Dr. Gottesman stated that the heat 
sensitive properties of the Streptcmyces spores are not convincing. Dr. Brill 
questioned the RAC as to the potential dangers the RAC forsaw if this 
proposal was approved. Dr. Gottesman responded that the RAC in the past 
had operated on the assumption that every possible danger could not be 
anticipated. Therefore, the RAC sought to contain recombinant DNA, and 
sporulating systems are not containable. The RAC previously preferred 
either high physical containment or limiting the kinds of DNA that were 
allowed to be cloned. She stated that at this time, there appears to be a 
change of philosophy on the RAC: if the RAC cannot see a specific danger, 
then the experiment will be approved. Dr. Gartland pointed out that the 
experiments Dr. Cohen requests approval for can be done under the current 
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