title: Overexpression of cardiac actin with baculovirus is promoter dependent.
abstract: The influence of the promoter and an N-terminal hexahistidine tag on human cardiac actin (ACTC) expression and function was investigated using four baculovirus constructs. It was found that both non-tagged ACTC and hisACTC expression from the p10 promoter was higher than from the polh promoter. Characterization showed that an N-terminal hexahistidine tag has a negative effect on ACTC. Recombinant ACTC inhibits DNase-I and binds myosin S1, indicative of proper folding. Our data support the hypothesis that the actin protein down-regulates the polh promoter.
authors.affiliation: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ont., Canada N1G 2W1.
authors.forename: Susan P
authors.initials: SP
authors.lastname: Yates
authors.forename: Marissa D
authors.initials: MD
authors.lastname: Otley
authors.forename: John F
authors.initials: JF
authors.lastname: Dawson
mesh_terms: D000199:Actins / Q000235:genetics*; D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002460:Cell Line; D005822:Genetic Vectors / Q000235:genetics*; D009206:Myocardium / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism*; D018411:Spodoptera / Q000235:genetics* / Q000378:metabolism* / Q000821:virology; D014162:Transfection / Q000379:methods*
chemical_list: D000199:Actins; D011994:Recombinant Proteins