Expression of the cp gene and Antiserum Preparation of Wisteria Vein Mosaic Virus
Abstract: The coat protein (CP) gene of the Beijing isolate of Wisteria vein mosaic virus (WVMV-BJ) was amplified by RT-PCR, and ligated to the expression vector pET22b(+). The recombinant plasmid pET-WVMVCP was transformed into E. coli BL21(DE3) and then induced by IPTG. It was shon that the CP gene was highly expressed by SDS-PAGE and Western blot analysis. The molecular weight of the recombinant protein was about 34.4kDa. Antiserum with high specificity was produced after the rabbit was immunized with purified recombinant protein, and the titer was determined to be 1/1024 by micro-precipitation, or 1/8192 by antigen coating plate- ELISA(ACP-ELISA).