23 
XIII. REQUEST TO UTILIZE HEMOPHILUS PARAINEIIJENZAE TO CLCMl MOLONEY MURINE 
I£UKEMIA PROVIRUS 
Dr. Bems began discussion of the proposal (tabs 1032, 1035/1, 1036) of 
Dr. James W. Gautsch of Scripps Clinic and Research Foundation to clone 
Molcxiey MuLV provirus and cellular flanking regions in Hemophilus para in- 
fluenzae . The provirus EWA and flanking regions will be ligated into vector 
pRK290, a plasmid with a broad host range in gram negative bacteria. The 
cloned plasmid will subsequently be used to infect NIH 3T3 cells. Dr. Bems 
said the investigators wish to study the effect of methylation of raiA on PNA 
transcription . 
Dr. Bems said |L parainfluenzae is part of the normal flora of the human 
upper respiratory tract. The investigators are thus inserting the MuLV 
provirus into a bacterium v^ich could colonize a laboratory worker. He 
noted, however, that Moloney MuLV virus is classified by the National Cancer 
Institute as a low risk virus. The normal host is the mouse, and the virus 
is not known to function in any other organism. Should the reoOTibinant ENA- 
containing parainfluenzae lyse in the respiratory tract of a colonized 
individual, the MuLV EWA would be presented to the cells of the respiratory 
tract as uncoated ES^JA, not as the whole virus. This is not the optimal 
manner in which to transfect cells. Dr. Bems said that he feels the risk is 
miniscule and recommended that the experiment be permitted at the P2 level of 
containment. 
Dr. Goldstein asked v^at the host range of the MuLV virus was in tissue cul- 
ture. Dr. Bems replied that MuLV is classified as an ecotropic virus, i.e., 
mouse-tropic. Dr. Goldstein asked how that classification was generated; was 
the test performed in tissue culture systems using whole virus? Dr. Berns 
replied that is was. 
Dr. Bems moved that the experiments be permitted at the P2 level of contain- 
ment. Dr. McKinney seconded the motion. By a vote of eleven in favor, none 
opposed, and three abstentions, the RAC adopted the motion. 
XIV. DEVELOPMENT OF HOST-VECTOR SYSTEM BASED CN CORYNEBACTERIUM GLUTAMICIM 
Dr. Maas introduced the request (tabs 1033, 1035/11) of Dr. Daniel Liberman of 
the Massachusetts Institute of Technology. Dr. Liberman requested containment 
conditions be established for the development of a host-vector syston based oi 
the gram positive bacteriun Corynebacterium glutamicum . Corynebacterium 
glutamicum would be used as the host; three types of plasmids including hybrid 
plasmids would be tested for use as vectors. Dr. Maas said Corynebacteriun 
glutamicum is not a pathogen and Pi containment should be adequate. 
Dr. Goldstein asked if the proposed plasmid vectors carry drug resistance 
genes. Dr. Maas replied that some did. Dr. Goldstein pointed out that 
although Corynebacterium glutamicum is not a pathogen, it is related to the 
organism causing diphtheria. 
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