The expression of hepatitis C virus truncated core and EGFP fusion gene in Sf9 cells / 临床检验杂志

ABSTRACT

Objective To construct recombinant baculovirus expression vector containing hepatitis C virus(HCV) truncated core gene and EGFP gene,and to study the antigenicity of the fusion protein expressed in Sf 9 cells.Methods PCR-amplified HCV truncated core gene and EGFP gene were cloned into the transposed vector pFastBac1 to construct a recombinant plasmid pFastCt-EGFP,by which E.coli DH10Bac was transformed to get the recombinant BacmidCt-EGFP.Insect Sf 9 cells were transfected with BacmidCt-EGFP and the expression of fusion protein Ct-EGFP was screened by SDS-PAGE and Western blotting.Results SDS-PAGE and Western blot analysis showed that fusion protein Ct-EGFP was expressed with the molecular mass of 40 kD.ELISA results showed that the fusion protein reacted with 15 of 28(54%) anti-HCV positive sera.Conclusion The fusion protein Ct-EGFP was expressed in insect Sf9 cells and showed partial antigenicity.