5 
I. CALL TO ORDER AND OPENING REMARKS 
Dr. Gartland introduced the newly appointed Chair of the Recombinant ENA 
Advisory Committee (RAC), Mr. Robert Mitchell. Mr. Mitchell called the 
meeting to order. He noted that a quorum was present, and that the meeting 
had been duly announced in the January 5, 1984, Federal Register (49 FR 
696). Mr. Mitchell then introduced the ad hoc consultants for the February 6, 
1984, meeting: Dr. Anne Vidaver of the University of Nebraska, Dr. LeRqy 
Walters of Georgetown University, Dr. Frances Sharpies of Oak Ridge National 
Laboratory, and Dr. Martin Alexander of Cornell thiversity. 
II. MINUTES OF THE SEPTEMBER 19, 1983, MEETING 
Mr. Mitchell called upon Mr. Daloz to review the minutes (tab 1142) of the 
September 19, 1983, meeting of the RAC. Mr. Daloz said he had reviewed the 
minutes and found them to be in order. He moved acceptance of the minutes 
as written. Dr. MoGonigle agreed and seconded the motion. Mr. Mitchell 
then called for a voice vote and the minutes were unanimously approved. 
III. PROPOSAL TO CLONE SHIGA-LIKE TOXIN FROM E. COLI 
Mr. Mitchell called upon Dr. Levine to present the proposal (tabs 1129, 
1130, 1131, 1132, 1133/1 II, 1137) of Dr. Alison O'Brien of the Uniformed 
Services University of the Health Sciences (USUHS) in collaboration with 
Dr. Randall Holmes (USUHS) to clone in Escherichia coli K-12 the structural 
gene of the Shiga-like toxin of E. coli~ Shiga-like toxin has activity 
similar to the activity of Shigella dysenteriae toxin. 
In a first submission in September 1982, the investigators preposed to 
clone the Shiga-like toxin gene in EL_ ooli EK1 host-vector systems using 
plasmid, cosmid, or lambda cloning vectors. In support of their proposal, 
Drs. O'Brien and Holmes offered the following arguments: 
(1) Clinical isolates of EL_ coli have already been demonstrated to 
elaborate large amounts of toxin indistinguishable from that 
produced by Shigella dysenteriae 1 (Shiga). Therefore, the genes 
for Shiga-like toxin production are present in the E. ooli gene 
pool found in nature. ““ “~~™ 
(2) Human volunteers fed large nurrbers of Shigella dysenteriae 1 
organisms that produced Shiga toxin but could not colonize the 
bowel did not become ill. Therefore, ary accidental ingestion of 
the organism to be manufactured, a toxin-producing EL coli K-12 
strain that cannot colonize the human intestinal tract, would pose 
little hazard to man. 
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