Expression of HCV CE2 fused protein in cultured cells and larvae of silkworm and initial application

    WANG Keª²Xia, WANG Xueª²Feng, PENG Jiangª²Long, LIANG Yong

    Department of Pathogenic Microbiology, School of Medicine, Anhui University of Science &Technology, Huainan 232001, China

    ¡¾Abstract¡¿ AIM: To highly express HCV CE2 fused protein and to study its initial application. METHODS: A recombinant transfer vector pBacPAKª²CE2, containing total cDNA of CE2 gene with 2a HCV strain, was constructed and coª²transfected into BmN cells with linearized BmBacPAK (modified BmNPV) DNA for the construction of a recombinant baculovirus carrying CE2 fused gene. The recombinants were identified by restriction endonucleases digestion and PCR amplification. After being infected by the recombinant baculovirus, all the samples of cell extract, culture supernatant, and hemolymph of larvae were detected for specific HCV CE2 bands by SDSª²PAGE. The biologic activity of the expressed product was detected with indirect ELISA. RESULTS: The genome of recombinant baculovirus contained a 1.6 kb inserted fragment identified by restriction endonucleases digestion and PCR amplification. From SDSª²PAGE analysis, a 90 ku band was determined in cell extract, culture supernatant and hemolymph of larvae. Indirect ELISA showed that the expression products had strong immunogenicity. CONCLUSION: HCV CE2 fused protein with biologic activity is highly expressed in cell culture and larvae of silkworm, which provides basis for further study on the vaccine against HCV infection and the development of diagnosis reagent. ......