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Virologica Sinica, 21 (6) : 536-540, 2006
Research Article
Expression of HBV X Gene and Its Function in Endoplasmic Reticulum Stress in Eukaryotic Cell Lines
State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, China
HBV X gene was amplified by PCR and cloned into the prokaryotic expressing vector pET-his and the eukaryotic expressing vector pcDNA3.1(-). E.coli BL21 (DE3) were transformed by the recombinant plasmid pET-his-HBx.. HBx protein was expressed by IPTG induction and formed insoluble inclusion bodies, which were dissolved in urea, dialyzed against PBS, and then applied onto Ni column. Anti HBx antibodies were generated in rabbits by immunization with the purified HBx protein. The recombinant plasmid pcDNA3.1(-)-HBx was transfected into HepG2 and Hep3B cell lines, in which HBx expression was detected by RT-PCR and Western blots. In this study, luciferase activities of XBP1 promoter and GRP78 promoter in HBx expressing HepG2 and Hep3B cells were increased by 3~7 folds compared with that of cells mock-transfected with pcDNA3.1(-). RT-PCR showed that XBP1 mRNA was spliced in HBx-expressing HepG2 and Hep3B cells compared with negative controls. Our results demonstrated that expression of HBx induced UPR in HepG2 and Hep3B cell lines and provided insights into the effects of HBx on ER and the mechanisms underlying liver pathogenesis.
  Published online: 20 Nov 2006
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