| abstract |
The present invention provides a novel, highly efficient, recombinant adenovirus expression system for expression of a heterologous gene(s) and/or gene product(s) in a mammalian cell. The recombinant adenovirus was produced by cotransfecting a novel vector with the large fragment of the adenovirus-5 genome in 293 cells. Homologous recombination between these two DNA fragments resulted in the production of the recombinant adenovirus expression system. This vector, when converted to a recombinant virus has the unique capability of expressing one or more heterologous genes at very high levels. The novel vector, comprises, at least one cDNA insertion site for cloning a selected heterologous gene; a promoter sequence positioned upstream from the gene insertion site; the left end replication and packaging elements of the adenovirus-5 genome positioned upstream of the promoter; a highly efficient eukaryotic splice acceptor and splice donor site positioned immediately downstream of the promoter; and positioned downstream of the insertion site a strong polyadenylation sequence and the region for homologous recombination containing a portion of the adenovirus-5 genome. Between the packaging sequence and the CMV promoter are restriction sites for insertion of a second fully functional transcription unit. |