CLONING SEGMENT SPIKE PROTEIN GENE OF SARS-COV AND ITSEXPRESSION IN ESCHERICHIA COLI

Objective Expressing and purifying the seglnent of SARS-CoV spike protein in E. Coli Methods The target gene was obtained by RT-PCR. The PCR product was cloned into pEGM- T Easy Vector, sequencing and double restriction digestion (BamH Ⅰ , Pst Ⅰ) were performed. The target gene was subcloned into PQE30 expression vector. The gene was expressed in the E. coli strain M15 cells induced by IPTG. The protein was purified with a nickel HiTrap chelating metal affinity column. Results The recombinant expression plasmid was successfully constructed and the protein was well expressed in E. coli strain M15 cells. The ideal pure protein was obtained by purification. Western blotting analysis suggested the protein could act with the convalescent sera of lab confirmed SARS patients. Conclusion The segment of SARS-CoV spike protein was well expressed and purified, and can be applied in diagnosis and immunological research of SARS.