Construction and proeukaryotic expression of a novel HBeAg binding protein 2 of hepatitis B virus in vitro

Objective To construct proeukaryotic expressive vector of HBEBP2 gene,and to observe the expression of recombinant protein in vitro. Methods The DNA fragment of HBeAg binding protein 2 was amplified by polymerase chainraction reaction,and cloned into pGEM-T vector. The constructed recombinant plasmid was identified by restriction analysis and DNA sequencing. The correct DNA fragment was inserted into inducible proeukaryotic expressive vector pET-32a(+) and transformed into E.coli BL21. The protein was induced with IPTG and analyzed with sodium dodecylsulfate-polyacrylamide gel electrophoresis and Western blot hybridization. Results The results of restriction analysis,PCR and DNA sequencing proved that HBEBP2 fragment was correctly inserted into vector pET-32a(+). After induction with IPTG,recombinant target protein with about Mr 34kD was expressed. Western blot analysis showed that the protein had good antigenicity. Conclusions The recombinant HBEBP2 gene was expressed successfully. These results lay the foundation for studying the immunogenicity and bionomics of the HBEBP2 protein.