Recombinant hepatitis B virus P22e gene in HepG2 cells.doc

 PAGE \* MERGEFORMAT 11 Recombinant hepatitis B virus P22e gene in HepG2 cells Author: Diao Zhihong, ZHANG Ming-xia, David Fu, He Begonia, Wang Zhan-Hui, Chen Jinjun, Hou Jinlin [Keywords:] Hepatitis B virus; gene recombination; eukaryotic expression Construction of recombinant vector of HBVP22e gene and its expression in HepG2 cells [Abstract] AIM: To construct the recombinant expression vector of pCDNA3.1 HBVP22e and transfect HepG2 cells for the expression of HBVP22e, and to assess its relations to HB. METHODS: HBVP22e cDNA was obtained from plasmid p3.8 Ⅱ which carried 1.2 copies HBV (adr subtype) genome by PCR and inserted into universal vector pMD18T for identification of its DNA sequence. The recombinant plasmids were constructed by pCDNA3.1 and transfected into HepG2 cells via liposom. HBVP22e protein was analyzed by immunohistochemistry and microparticle enzyme immunoassay. RESULTS: Expression vectors of recombinant pCDNA3.1 HBV P22e were successfully constructed and transferred into HepG2 cells. Its expressed steadily in HepG2 cells after cultured over several passages. CONCLUSION: HBVP22e can be expressed in HepG2 cells. [Keywords] hepatitis B virus; gene recombinant; eukaryote expression [Abstract] Objective: Using the pCDNA3.1 eukaryotic expression vector and established stable expression of hepatitis B (HBV) P22e the HepG2 cell line to study the pathogenesis of hepatitis B HBVP22e relationship. Methods: containing 1.2 copies HBV gene (adr subtype) p3.8II plasmid as a template, PCR was HBVP22e cDNA fragments, separation into common T vector pMD18T identified, and then into the eukaryotic expression vector pCDNA3.1, the lipofection HepG2 cell lines Immunohistochemical identification of cells expressed CMIA check the supernatants secreted antigen. RESULTS: The successfully constructed the eukaryotic expression vector pCDNA3.1 HBVP22e, and transfected HepG2 cells after repeated subculture identification, to obtain stable expression of HBVP22e H