Vector ConstructionHuman Akt-Myr cDNA with myristolation signal for greater activity, also known as PKB, 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 human Akt cDNA expression cassette, generating a replication deficient recombinant virus.
Supplied AsCrude HEK293 cell lysate.
Storage Store at -80°C. Multiple freeze/thaw cycles not recommended.
QC TestingWestern Blot
Recombinant adenovirus was generated as described in Gomez-Foix, AM, et al., J. Biol. Chem. (1992)267:25129-25134.