May 21-23 - MINUTES OF MEETING 
21 
"II-!>l-a-( 1) . EK1 . The host is always E. coli 
K-12 or a derivative thereof, and the vectors 
include nonconjugative plasmids (e.g., pSClOl, 
ColEl, or derivatives thereof [21-27] ) and 
variants of bacteriophage, such as [28-33] . 
When plasmid vectors are employed , the E. coli 
K-12 hosts shall not contain conjugation- 
proficient plasmids, whether autonomous or 
integrated, or generalized transducing phages." 
This proposal is an outgrowth of discussions on the use of M13 as a vector 
at the last RAC meeting. Its effect is to restrict the prohibition against 
cloning in conjugation-proficient hosts to plasmid vectors and to exclude 
phage vectors frcm the prohibition except for those phages which are pro- 
pagated as plasmids. The currently approved phage vectors are the filamentous 
single-strand DNA phages (Ff phages) and the lambda-related (lambdoid) 
phages. Dr. Gottesman elaborated on the definition of EK1, and emphasized 
that conjugative plasmids and generalized transducing phages are not 
permitted in EK1 systems. She pointed out that M13 has been approved in 
experiments involving DNA transfection, and the use of transfer-minus 
plasmids. 
Several RAC members expressed reservations about permitting the use of 
conjugative plasmids and generalized transducing phages. It was pointed 
out that the M13 system is working well in the presence of transfer-minus 
plasmids. The RAC rejected the proposed amendment by a vote of 10 to 4, 
with 5 abstentions. 
XV. CLONING IN BACILLUS SUBTILIS AND STREPTOMYCES COELICOLOR 
Dr. Broadbent summarized a request frcm Dr. Stanley Cohen of Stanford 
University. In the first request. Dr. Cohen preposed that Bacillus 
subtilis strains that do not carry an asporogenic mutation can be used 
as hosts specifically for the cloning of DNA derived frcm EL coli K-12 
and Streptcmyces coelicolor using NIH-approved S taphylococcus aureus 
plasmids as vectors under P2 conditions. It was noted that this proposal 
requests the use of B. subtilis strains that do not carry an asporo- 
genic mutation, whereas the NIH-certif ied B. subtilis HV1 system requires 
the use of an asporogenic mutant derivative of B. subtilis as the host 
ccmponent. It was also noted that P2 containment is preposed for these 
experiments, whereas section III-Br-3 of the Guidelines, as amended on 
April 11, 1979, requires the use of P3 containment for experiments between 
bacterial species that are nonpathogens and for which genetic exchange has 
not been demonstrated. 
[ 116 ] 
