Strategy outline. To generate E1 and E4 heterologous promoter controlled replication-competent adenoviral vectors, the promoters of interest are inserted into the appropriate multiple cloning sites (MCS) of the transfer vector pShuttle#4-1 (or pShuttle#4-1Δ24). E. coli BJ5183 cells are transformed with the adenoviral backbone plasmid pAdEasy-1 or pTG3602 and the linearized transfer vector. Recombinants resistant to kanamycin are analyzed by restriction enzyme analysis to determine whether homologous recombination occurred at the corresponding "left arm" and "right arm". Subsequently infectious recombinant adenovirus is rescued from 293 cells.