COMPLETE SEQUENCE AND TRANSPOSON MUTAGENESIS OF THE BAMHI J-FRAGMENT OF CYDIA-POMONELLA GRANULOSIS-VIRUS

The BamHI J fragment of Cydia pomonella granulosis virus was subcloned and subjected to transposon mutagenesis in Escherichia coli using a T n3 derivative. After screening by restriction endonuclease digestion a nd polymerase chain reaction (PCR), 44 clones were selected representi ng insertions every 100 to 300 bp. The complete sequence was compiled and the transposon insertion sites mapped precisely by sequencing. Ana lysis of the sequence revealed the presence of 7 potential open readin g frames (ORFs). The BamHI J fragment was already known to encode IAP and OPDV-E6. Three other ORFs encode products similar to known protein s, viz. an Autographa californica nuclear polyhedrosis virus 8.6 kDa p rotein, a Lymantria dispar nuclear polyhedrosis virus 34 kDa protein, and vertebrate reovirus sigma-1 proteins. The ORF with similarity to s igma-1 is also similar to baculovirus p10 proteins. In both cases, the similarity occurs in regions likely to form a coiled-coil structure.