We have recently cloned intact duplex adeno-associated virus DNA into the bacterial plasmid pBR322 (1, Appendix). When the recombinant plasmid, pSM was transfected into human cells with Ad5 as helper virus, the AAV sequences in the plasmid were rescued, replicated, and packaged into virions. The efficiency of rescue from the plasmid was sufficiently high to produce yields of AAV DNA comparable to those observed after transfection of equal amounts of purified varion DNA. We propose to 1) evaluate AAV as a possible mammalian DNA cloning vector, 2) study the mechanism of rescue from the recombinant plasmid, and 3) construct a preliminary complementation map of the AAV genome.
