Overexpression of cardiac actin with baculovirus is promoter dependent

Susan P Yates; Marissa D Otley; John F Dawson

doi:10.1016/j.abb.2007.07.018

Overexpression of cardiac actin with baculovirus is promoter dependent

Arch Biochem Biophys. 2007 Oct 1;466(1):58-65. doi: 10.1016/j.abb.2007.07.018. Epub 2007 Aug 6.

Authors

Susan P Yates¹, Marissa D Otley, John F Dawson

Affiliation

¹ Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ont., Canada N1G 2W1.

PMID: 17765196
DOI: 10.1016/j.abb.2007.07.018

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Actins / genetics*
Animals
Baculoviridae / genetics*
Cell Line
Genetic Vectors / genetics*
Myocardium / metabolism*
Promoter Regions, Genetic / genetics*
Protein Engineering / methods*
Recombinant Proteins / metabolism*
Spodoptera / genetics*
Spodoptera / metabolism*
Spodoptera / virology
Transfection / methods*

Substances

Actins
Recombinant Proteins