POTENTIAL SALMON SPERM ORIGIN OF THE E3 REGION INSERT OF THE ADENOVIRUS-5 DL309 MUTANT

The plasmid pJM17 is a commonly used adenoviral backbone derived from the dl309 mutant virus. It contains unknown sequences inserted in the E3 region during construction of the dl309 mutant. Complete descriptio n of the backbone sequence is required for interpretation of potential vector effects and for regulatory approval of a vector to be used in clinical trials. The anonymous E3 insert was sequenced and analyzed. T he insert fragment is 646 base pairs (bp) long and is 100 bp shorter t han the vector sequences it replaces. It interrupts the expression of the E3B 10.4K, 14.6K, and 14.7K genes, but not the E3A glycoprotein (g p)19K gene. Sequence analysis and Southern blotting suggest that the i nsert might originate from salmon sperm DNA used as carrier during the construction of dl309. Transcription from the insert was not detected by Northern blot analysis of vector-transduced cells but was detected by reverse transcriptase polymerase chain reaction.