doi: 10.1016/s1046-5928(02)00649-6.
Construction and purification of his6-Thermus thermophilus MutS protein
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- PMID: 12651109
- DOI: 10.1016/s1046-5928(02)00649-6
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Construction and purification of his6-Thermus thermophilus MutS protein
Protein Expr Purif.
2003 Mar.
Abstract
The mutS gene from the thermophilic bacterium Thermus thermophilus was PCR amplified, cloned, and expressed in Escherichia coli. The recombinant MutS protein containing an oligohistidine domain at the N-terminus was purified in a single step by Ni(2+) affinity chromatography to apparent homogeneity. The mismatch recognition properties of the his(6)-tagged MutS protein were confirmed by DNA protection against exonuclease digestion and retardation assays. The results of analytical gel filtration indicate that the predominant form of T. thermophilus MutS at micromolar concentrations is a tetramer.
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