Cloning and prokaryotic expression of HBV Wholeª²X gene

    XUE Hongª²An, YANG Qian£¬ LIU Laª²Yang, CHENG Jun, DONG Jing

    1Department of Infection, Second Hospital, Xi¡¯an Jiaotong University, Xi¡¯an 710004, China, 2Gene Therapy Research Center, Institute of Infectious Disease, 302 Hospital of Chinese PLA, Beijing 100039, China

    ¡¾Abstract¡¿ AIM: To construct the prokaryotic expression plasmid pETª²32a(+)ª²Wholeª²X. METHODS: The Wholeª²X gene was cloned into EcoR I/Sal I site of pETª²32a(+) and was sequenced. The fusion protein was expressed in E. coli with IPTG induction. After the induction and purification, the protein was analyzed in SDSª²PAGE and identified by Western blotting. RESULTS: Endonucleases digesting and sequencing confirmed that the Wholeª²X gene was correctly inserted into the vector. The Wholeª²X gene was successfully expressed in E.coli. The fusion protein was confirmed by SDSª²PAGE and Western blotting. CONCLUSION: The prokaryotic expression plasmid pETª²32a(+)ª²Wholeª²X has been successfully reconstructed, which provides the experimental base for study on its biological functions and antibody preparation. ......