3 
Dr. Friedman said currently most experiments with gram-positive organisms not 
covered by Appendix A are covered by Section III-C. This section requires 
investigators to register with the local Institutional Biosafety Camuttee 
(IBC) at initiation of the experiments. The experiments can then proceed 
under Biosafety Level (BL) 1 containment conditions. Experiments involving 
Class 2 pathogens are covered by Section III-B, Experiments Which Require IBC 
Approval Before Initiation of the Experiment . 
Dr. Friedman suggested the working group begin by examining the proposal 
(Attachment II) submitted by Drs. Novick and Polak. 
Dr. Baltz asked whether the working group could first discuss the relevance of 
the criteria used for adding organisms to the sublists of Appendix A. 
Dr. Clewell noted that the NIH Guidelines have been constructed in accordance 
with knowledge of E^ ooli genetics. Dr. Dean questioned whether criteria 
based on the genetics of gram-negative bacteria are appropriately applied to 
gram-positive bacteria. Exchange of DNA in gram-positive bacteria does not 
follow the model observed in gram-negative bacteria. Gram-positive bacteria 
do not have the same type of oonjugative plasmids observed in EL_ coli or the 
mechanisms of genetic exchange observed in gram-negative bacteria. He thought 
plasmid exchange alone, whether or not it mobilizes chromosomal exchange, 
should be adequate evidence for including organisms on the sublists of 
Appendix A. 
Dr. Lovett said in gram-negative bacteria, F' episomes can be generated in the 
laboratory and introduced into bacteria that would not otherwise exchange DNA. 
Since F ' episomes are not known in gram-positive bacteria and chromosomal gene 
transfer by this type of conjugation has not been described in these organisms, 
the same test cannot be applied. It is likely, however, that F' episomes 
exist in gram-positive bacteria. 
Dr. Baltz supported Dr. Dean's argument; he felt evidence of plasmid transfer 
should suffice for including organisms on Appendix A. 
Dr. Friedman said the working group can offer other criteria for exemption 
to the Director, NIH, but whatever recairnendation the working group offers 
should be supported by strong scientific arguments. 
Dr. Thcmashcw said the working group is to evaluate the probability that the 
combination of genes constructed in the laboratory using recombinant DNA 
techniques might occur naturally. The exchange of chromosomal markers must 
be addressed if organisms are to be included on an Appendix A sublist on the 
basis of "non-novelty" of the exchange. Hew transfer occurs is not an issue, 
and the working group need not be concerned with the mechanism. 
[534] 
