Overexpression of cardiac actin with baculovirus is promoter dependent.
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.
Yates
Susan P
SP
Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ont., Canada N1G 2W1.
Otley
Marissa D
MD
Dawson
John F
JF
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*
D000199:Actins; D011994:Recombinant Proteins