Construction and Expression of Eukaryotic Vector for HBV X

Objective This study was designed to construct and express the eukaryotic expression vector of HBV X gene for exploring the contribution of X gene to the chronic hepatitis and the hepatocellular.Methods X gene with EcoRⅠand Hind Ⅲ endoenzyme sites was obtained by using PCR,and subcloned into pcDNA3.1(+) vector.After identified by restrictive enzymes digestion and sequencing,reconstructed plasmid was transfected into THP-1 cells.Expression of X were assayed in THP-1 cell lysate by Western-blotting.Results The target gene X fragment about 465 bp was obtained.In THP-1 cell,the pcDNA3.1(+)-HBx plasmid expressed X protein with molecular weight of 17 kDa by Western-blotting.Conclusion Eukaryotic expression vector(pcDNA3.1)(+)-HBx was constructed successfully and X protein was expressed in THP-1 cell.