25 
MINUTES OF MEETING - June 5-6 
acceptance of the proposed experiments at P2 containment. The 
RAC recommended approval of Dr. Sypherd's proposal by a vote of 
fourteen in favor, none opposed, and four abstentions. 
XVII. PROPOSED FUTURE MAJOR ACTIONS 
A. Proposal to Introduce Arabidopsis DNA Ligated into an Escherichia 
coli/ri Hybrid Plasmid into Arabidopsis 
Dr. Gottesman said that Dr. Donald Merlo of the University of Missouri- 
Columbia had requested (tab 892) advice from the RAC on the containment 
appropriate to the following protocol: (1) clone the thiamine (thi) 
genes from E^ coli or Agrobacterium tumefaciens in a thi" E. coli 
K-12; (2) clone the origin of replication, and possibly thi~~bacteria- 
plant transfer genes of the Ti plasmid of Agrobacterium tumefaciens 
in E_j_ coli K-12; (3) clone Arabidopsis DNA fragments in the pBR325/Ti 
plasmid (PL2) in step two in E. coli K-12 or xl776; (4) isolate the 
thiamine gene from the pBR3257thi plasmid (PL1) in step one, and 
insert them by restriction and ligation into the Arabidopsis DNA 
sequences in the pBR3 2 5/Ti /Arabidops is plasmid (PL3) from step three; 
(5) the reisolated PL4 plasmid would be transformed into Agrobacterium 
tumefaciens ; and (6) infect thiamine mutants of Arabidopsis with 
Agrobacterium tumefaciens containing PL4. 
Dr. Zaitlin said the first four steps are covered by Section III-O 
of the Guidelines. He noted that Arabidopsis can be grown in a test 
tube and can be easily contained. It was agreed that steps 5 and 6 
were covered by Appendix E and that containment should be P3. 
Dr. Gottesman suggested that as the Escherichia coli and Agrobacterium 
tumefaciens DNA fragments are well characterized , and Arabidopsis 
DNA will be returned to Arabidopsis , containment for these particular 
experiments could be P2. She moved approval of steps 5 and 6 at P2 
containment. A vote of eleven in favor, none opposed, and five 
abstentions supported Dr. Gottesman' s motion. (Executive Secretary's 
Note: Dr. Merlo 's request will be reconsidered at the next RAC 
meeting following publication in the Federal Register . ) 
B. Review of Section I-D of the Guidelines 
Dr. Nightingale said that an ad hoc group had been formed following 
the March 6-7, 1980 meeting to determine whether Section I-D of the 
Guidelines required revision. Their report is tab 894. 
Dr. Nightingale noted that Section I-D-l, prohibiting formation of 
recombinant ENAs derived from CDC Class 3, 4 or 5 agents, had been 
recommended for modification at this meeting so as to permit experi- 
ments with Class 3 agents while still prohibiting use of Class 4 and 
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