Ser 189/193 Ala mutant in MAPK kinase 3 human cDNA with 5’ flag epitope was cloned into the multiple cloning site of pAC-CMV-pLpA. The resulting plasmid was cotransfected into HEK293 cells with plasmid pJM17 which contains the Ad5 genome.   Homologous recombination between the two plasmids resulted in the replacement of the Ad5 early region 1 with the MKK3, human cDNA expression cassette, generating a replication deficient recombinant virus. 

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