Title: Expression of human exCAR protein in E.coli and its purification and identification
Abstract: Objective To express the extra-cellular region of human Coxsackie-adenovirus receptor(exCAR) in E.coli,to purify the protein and identify it by N-end sequencing.Methods The gene fragment coding the extra-cellular region of CAR was amplified from the cDNA library of Hela cell strain,and linked into the expression plasmid pET-28a(+) to construct the expression plasmid pET-28a(+)-exCAR.After identified by restrictive enzyme cut and DNA sequencing,the expression plasmid was transformed into E.coli BL21 strain,the high expression strain was selected and induced,the expressed recombinant protein was purified by using Ni+ affinity chromatograph,and the purified protein was transferred to PVDF membrane after SDS-PAGE,then identified by Western-blotting.Results The expressed recombinant protein was purified,it was expressed as include body form,and the expressed protein was identified to be human exCAR protein by Western-blotting.Conclusion The exCAR recombinant protein was expressed and purified,and this work provides a basis and materials for the following adenovirus-blocking experiment in vitro.
Publication Year: 2009
Publication Date: 2009-01-01
Language: en
Type: article
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