Vector ConstructionReporter adenovirus for MEF-2 DNA binding activity and transcriptional activation multimerized MEF2 sites upstream of a minimal TATA-containing promoter was fused to a luciferase cDNA cassette (the CMV promoter was removed). It was cloned into the multiple cloning site of pAC-CMV-pLpA from which the CMV promoter was removed. The resulting plasmid was cotransfected into HEK293 cells with plasmid pJM17 containing the Ad5 genome. Homologous recombination between the two plasmids resulted in replacement of Ad5 early region 1 with MEF-2 DNA expression cassette, generating a replication deficient recombinant virus.
Supplied AsCrude HEK293 cell lysate.
StorageStore at -80°C. Multiple freeze/thaw cycles not recommended.
Wilkins BJ, Dai Y-S, Bueno OF, Parsons SA, Xu j, Plank DM, Jones F, Kimball TR, Molkentin JD. Calcineurin/NFAT coupling participates in pathologic, but not physiologic cardiac hypertrophy. Circ. Res. 94:110-118 2004.
Recombinant adenovirus was generated as described in Gomez-Foix, AM, et al., J. Biol. Chem. (1992)267:25129-25134.