[Cloning, Sequence Analysis and Expression of Recombinant E2 Protein of GB Virus C Genotype 7].

Abstract

The purpose of this study was to explore the potential of the GB virus C(GBV-C)genotype 7E2 protein as a detection antigen for ELISA kit development. In this study, analyses of antigen epitopes, space structures and the linear B cell epitopes from the GBV-C genotype 7E2 protein were performed using an online analysis program. To establish a more reliable detection method for GBV-C studies, a 945bp gene fragment from GBV-C E2 was amplified by RT-PCR and ligated into the pET-32 a prokaryotic expression vector, which was then transformed into E. coli BL21 cells for protein expression. A protein with a molecular weight of 55 kDa was detected by 12% SDS-PAGE. The protein was found in inclusion bodies, and the His-tagged protein was detected by western blotting. The results showed that the cloned E2 gene sequence was 945 bp, and that the GBV-C E2 protein sequence had multiple antigenic epitopes. The recombinant protein formed inclusion bodies, which was consistent with expectations. These findings may provide the foundation for the development of a GBV-C detection kit.