Construction of eukaryotic expression p1asmids of HBeAg and its expression in Bewo cells in Vitro

【Objective】To construct the recombinant eukaryotic expression vector pcDNA3.1(+)-HBe,and observe its expression in Bewo cells in vitro. 【Methods】Amplifying the HBeAg gene from the pMD18T-HBV vector by PCR,the product of PCR was cloned into the pcDNA3.1 (+),the recombinant vector was confirmed by restriction enzyme digestion,PCR and sequencing,then it was transfected into Bewo cells. After 72 hours,The extracellular and intracellular HBeAg expression was detected by Western blotting and Microparticle Enzyme Immunoassay (MEIA)【.Results】The results of the restriction enzyme digestion,PCR and sequencing confirmed the vector was constructed successfully,furthermore it can express HBeAg protein in Bewo cells and secrete it in supernatant. 【Conclusions】The vector is constructed successfully,which establishes the foundation for future research on the effect of intracellular HBeAg on the expression of Toll-like receptors in Bewo cells.