Vector ConstructionDominant negative PKC-alpha (protein kinase C) mutant cDNA 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 PKC alpha-DN mutant cDNA expression cassette, generating a replication deficient recombinant virus.
Supplied AsCrude HEK293 cell lysate.
StorageStore at -80°C. Multiple freeze/thaw cycles not recommended.
QC TestingWestern Blot
Braz JC, Bueno OF, De Windt LJ, Molkentin JD. Protein kinase C? (PKC?) regulates the hypertrophic growth of cardiomyocytes through extracellular signal-regulated kinase1/2(ERK1/2). J. Cell Biol. 156:905-919 2002.
Recombinant adenovirus was generated as described in Gomez-Foix, AM, et al., J. Biol. Chem. (1992)267:25129-25134.