Exogenous gene expression in recombinant adenovirus for minimal impact on viral kinetics

A recombinant adenovirus genome, comprising a heterologous open reading frame (ORF) and a self-cleaving peptide coding sequence, both operably linked to and in the same reading frame as an endogenous adenovirus ORF, wherein the self-cleaving peptide coding sequence is located between the heterologous ORF and the endogenous ORF, and wherein:

• the endogenous ORF is E1B-55k and the heterologous ORF is 3′ of E1B-55k;
• the endogenous ORF is DNA polymerase and the heterologous ORF is 5′ of DNA polymerase;
• the endogenous ORF is DNA-binding protein (DBP) and the heterologous ORF is 3′ of DBP;
• the endogenous ORF is adenovirus death protein (ADP) and the heterologous ORF is 5′ of ADP;
• the endogenous ORF is E3-14.7k and the heterologous ORF is 3′ of E3-14.7k;
• the endogenous ORF is E4-ORF2 and the heterologous ORF is 5′ of E4-ORF2;
• or
• the endogenous ORF is fiber and the heterologous ORF is 3′ of fiber.