Construction of the Eukaryotic Expression Vector of HCV NS3-5B and Its Expression in BHK-21 Cells

To construct the recombinant plasmid pIRES2-EGFP-NS3-5B,HCV NS3-5b gene was amplified by the polymerase chain reaction( PCR) and cloned into the eukaryotic expression plasmid pIRES2-EGFP. Then the recombinant plasmid was transfected into BHK-21 cells by Lipofectamine 2000 and the expression of target genes were identified by fluorescence imaging and western blot analysis. The results showed that the plasmid pIRES2-EGFP-NS3-5b was successfully constructed and NS3 /4A and NS5B proteins were specifically expressed in the eukaryotic cells. It provides the basis for the establishment of a cell-based HCV RdRp evaluation system and animal models development.