Simultaneous insertion of two expression cassettes into adenovirus vectors

We have designed AdenoQuick, a fast and versatile method to construct first -generation adenoviral vectors that contain one or two transgenes in the E1 and/or the E3 region. The method is based on the reconstitution of the ent ire genome of the desired recombinant vints in E. coli and the subsequent t ransfection of the DNA in a helper cell line Since the construction of larg e adenoviral plasmids is generally difficult and therefore rebuffing for in experienced researchers, we have optimized the cloning strategy by rising b acterial positive-selection markers and a set of specific restriction enzym es that allow for directional cloning. The system is 99% efficient and allo ws one to insert simultaneously two expression cassettes into the El and E3 regions of the adenovirus genome.