3 
Or. Oottesman said Appendix F-IV-H of the NIH Guidelines currently reads: 
"'The intact structural qene(s) of the Shiga-like toxin fran E. coll 
may be cloned in FL coll K-12 under RL3 + EK1 contairment conditions. 
" F. ooli host vector systems expressing the Shiga-like toxin gene may be 
moved fran RL3 to RL2 contairment conditions provided that: (1) the 
amount of toxin produced by the modified host- vector systems is no 
qreater than that produced by the positive control strain 933 E. coll 
0157:H7, grown and measured under optimal conditions; and (2) the cloning 
vehicle is to be an EK1 vector preferably belonging to the class of 
poorly mobilizable plasmids such as pER322, pER328, and pBR325. 
"Nontoxmogenic fraaments of the Eh iq a- like toxin structural gene(s) maybe 
moved fran RL3 + FK1 to RL2 + EK1 contairment conditions or such nontoxino 
qenic fragments may be directly cloned in FL coll K-12 under BL2 + EK1 
conditions provided that the EL_ coll host-vector systems containing the 
fragments do not contain overlapping fragments which together would encan- 
pess the Eh ip a- like toxin structural gene(s)." 
Drs. Habig and Levine said they might have conflict of interest considerations 
in dealing with the proposal fran Dr. O'Brien as they both have collaborative 
associations with her. Dr. Gottesnan suggested they participate in the discus- 
sion and vote on motions. Dr. Milewski would note their votes and the potential 
conflict of interest in the minutes of the meeting. 
Dr. O'Brien said she and her group had been investagating organisms for the 
presence of SLT. SLT is defined as being cytotoxic to cells and antigenically 
cross- reactive with purified FL coll SLT. SLT has been identified in members of 
the tallies Enterobactenaceae and the Vibnonaceae , and she was now requesting 
permission to clone SLT genes fran the Vibnonaceae and the Enterobactenaceae in 
E. coll K-12. The major qoal of this work is to develop a cholera toxin vaccine. 
Dr. O'Brien suggested SLT may be a common property of these bacteria; in 
organisms such as Shigella dysentenae 1 SLT may play a role in virulence when 
a large amount of the protein is produced and other virulence factors cure 
present . 
Dr. O'Rnen said .SLT has also been recently identified m one strain of Campylo - 
bacter and two strains of Aercmonas. The Aeromonas are classified as 
Vibnonaceae . The Campylobacter had at one time been classified by Bergey's 
Manual as Vibnonaceae but had recently been reclassified as an orphan species. 
Dr. O'Brien pointed out that a number of Campylobacter characteristics are 
camon to the Vibnonaceae . She asked whether she might receive permission to 
clone SLT qenes from Canplyobacter . 
Dr. Milewski said the August IB, 19B5, Federal Register announcement (50 FIR 
T3462) pub! ished TO days before the RAC meeting and describing the proposal 
stated that Dr. O'Brien renuestad permission to clone the SLT gene from members 
of the families Enterobactenaceae and V ibr lonaceae as classified m Bergey’s 
Manual . She sunqested the working qroup first vote on the proposal as it 
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