Chinese wheat mosaic virus(CWMV)is a soil-borne virus discovered and identified by Zhejiang Academy of Agricultural Sciences,whose genomic segment RNA2 encodes a major coat protein(CP)that plays an important role in virus infection.To detect its expression and analyze its biological function,the gene was amplified by RT-PCR from CWMV-infected wheat leaves.The recombinant expression vector was constructed with In-Fusion method and intro-duced into BL21(DE3)strain for the induction of CP expression.The recombinant CP protein was purified by nickel column affinity chromatography and injected into the rabbits for immunization.The antisera were purified by affinity chromatography and the polyclonal antibodies against CP were obtained.Dot-ELISA,ID-ELISA and Western blot as-says showed that the purified antibodies were highly specific to CWMV with a titer of 1∶2.048 × 107 and the sensitivity of 6.25 × 10-2 ng.Dot-ELISA and Western blot assays showed that the antibody could be used for accurate diagnosis of CWMV,which laid the foundation for CWMV detection,CP quantitation and its functional research.
关键词
中国小麦花叶病毒/外壳蛋白/重组蛋白/多克隆抗体
Key words
Chinese wheat mosaic virus/coat protein/recombinant protein/polyclonal antibody
维普
万方数据