[Preparation of recombinant IL-1 fusion protein as the substrate for HCV NS3 serine protease].

AIM: To prepare recombinant IL-1 fusion protein as the substrate for the HCV NS3 serine protease. METHODS: NS5A-B gene fragment (2 412-2 427aa) synthesized by PCR was subcloned into prokaryotic expression vector pBVIL1 to fuse with IL-1 gene and the recombinant vector pBVIL1/NS5A-B was transformed into E.coli strain HB101. The fused protein was induced to express at 42℃ and purified by two-step column chromatography. The proteolysis of the purified IL-1 fusion protein catalyzed by NS3 serine protease was analyzed with SDS-PAGE, Western blot and ELISA. RESULTS: NS5A-B fragment gene was correctly subcloned into pBVIL1 vector and the fusion protein was expressed as inclusion body in transformed HB101 cells. The recombinant fusion protein can be cleaved into smaller fragments by NS3 protease. CONCLUSION: The recombinant fusion protein can be cleaved by NS3 serine protease successfully and specifically, suggesting that it can be used as a surrogate substrate of NS3 serine protease in searching for inhibitors of this protease.[