of tissues and PCR analysis of DNA from testes or ovaries. 
Although an autopsy cannot be required as a precondition for 
entry into this study, this issue will be discussed with each 
patient. If possible, such analyses will be performed on all 
patients. This has been described in the revised protocol 
(Section 6.5). 
3. Patients will be sensitized to the HLA-B7 antigen likely 
making them ineligible for transplantation with organs from 
donors with this antigen (~15% of donors) . This information has 
been included in the revised "Informed Consent" (Section 15) . 
4. The vector sequence, together with all potential open 
reading frames, have been provided on computer disk. No open 
reading frames encoding oncogenes have been detected, and these 
open ready frames are shown (Appendix V) . In addition, this 
vector has been used in 3T3 transformation assays, and does not 
cause cell transformation. Although three vectors have been 
prepared and characterized, vector II is expressed best both in 
human melanomas and other cells and is proposed for use in this 
study. 
5. Clarification of the integration status of the plasmid 
DNA was requested. In general, we find that plasmids which are 
not linearized prior to lipofection are expressed transiently, in 
contrast to linearized plasmids which stably integrate into host 
cells with high frequency. We are currently performing Southern 
blot analysis of episomal or chromosomal DNA to determine the 
percentage of plasmids which remain episomal or integrate after 
lipofection. 
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Recombinant DNA Research, Volume 15 
