Vector ConstructionWild-type cMyBP-C cDNA was subjected to site directed mutagenesis in which the PKA/CaMKII/PKC sites (Ser-273, -282 and -302) were mutated to Alanine (cMyBP-CPhos-mutant) to mimic constitutive dephosphorylation (phospho-ablation). cMyBP-CPhos-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 mouse cDNA expression cassette, generating a replication deficient recombinant virus.
Supplied AsCrude HEK293 cell lysate.
StorageStore at -80°C. Multiple freeze/thaw cycles not recommended.
Epitope TagA myc-tag EQKLISEEDL is placed after the initiator Met (first ATG) codon
QC TestingWestern Blot - see datasheet
Cardiac myosin-binding protein-C phosphorylation and cardiac function. Circ Res. 2005 Nov 25;97(11):1156-63. Recombinant adenovirus was generated as described in Gomez-Foix, AM, et al., J. Biol. Chem. (1992) 267:25129-25134.