Gene ASK1-dominant negative
Vector ConstructionHuman cDNA for apoptosis signaling kinase 1 with a mutation in catalytic lysine (presumed dominant negative). It 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 mouse cDNA for ASK1-KM, generating a replication deficient recombinant virus.
Supplied AsCrude HEK293 cell lysate.
StorageStore at -80°C. Multiple freeze/thaw cycles not recommended.
QC TestingWestern Blot
Liu Q, Wilkins BJ, Lee YJ, Ichijo H, Molkentin JD. Direct interaction and reciprocal regulation between ASK1 and calcineurin-NFAT control cardiomyocyte death and growth. Mol Cell Biol. 2006;26:3785-97.
Recombinant adenovirus was generated as described in Gomez-Foix, AM, et al., J. Biol. Chem. (1992)267:25129-25134.