PMID:12561194

Citation	Bi, LJ, Zhou, YF, Deng, JY, Zhang, XE, Zhang, CG and Cass, AE (2002) [High expression and identification of DNA mismatch repair gene mutS in Escherichia coli]. Sheng Wu Gong Cheng Xue Bao 18:536-40
Abstract	DNA mismatch repair gene mutS (2.56 kb) was PCR modified and cloned into a secretive prokaryotic expression vector pET32a (+) which carries a N-terminal His.tag + and thioredoxin sequence. MutS protein was expressed with high level after IPTG induction using the strain E. coli AD494(DE3). SDS-PAGE revealed that the expected protein with a molecular weight of 108 kD which is about 35% of the total bacterial proteins is almost soluble. The expected protein was purified directly by immobilized metal (Ni2+) chelation affinity chromatography and the purity is over 90%. MutS protein activity verified using mismatch DNA showed that the expression product can recognize and bind to base-pair mismatch specifically.
Links	PubMed
Keywords	Adenosine Triphosphatases/biosynthesis; Adenosine Triphosphatases/genetics; Adenosine Triphosphatases/isolation & purification; Bacterial Proteins; Base Pair Mismatch; Chromatography, Affinity; DNA/metabolism; DNA Repair; DNA-Binding Proteins; Escherichia coli Proteins/biosynthesis; Escherichia coli Proteins/genetics; Escherichia coli Proteins/isolation & purification; Magnesium/pharmacology; Molecular Weight; MutS DNA Mismatch-Binding Protein; Recombinant Proteins/biosynthesis
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