Construction,expression and identification of fusion protein Tat_(47-57)-HBcAg in E. coli

Objective:To construct recombinant plasmid with fusion gene Tat47-57-HBcAg,express fusion protein Tat47-57-HBcAg in E.coli and identification the Tat47-57-HBcAg by western blot analysis.Methods:To synthesize the sequence of Tat47-57 and amplify HBcAg gene by PCR,splice the two sequences with splicing by overlap extension PCR,link fusion gene into pET28a,the sequences correct vector be transformed into E.coli Rosetta-gamiTM 2(DE3),then the transformed E.coli is induced by isopropyl β-D-1-Thiogalactopyranoside and the expression product is analyzed by SDS-PAGE,furthermore,identification the expression product by Western blot with HBcAg monoclonal antibody.Results:Fusion protein Tat47-57-HBcAg is highly effective expressed in E.coli Rosetta-gamiTM 2(DE3).The solubility analysis of expression product indicated that the fusion protein are mostly expressed in supernatant and expression product could react with HBcAg monoclonal antibody by western blot analysis.Conclusions:The Tat47-57-HBcAg fusion protein can be expressed in E.coli Rossetta-gami 2 correctly as soluble protein and be recognized by HBcAg monoclonal antibody.