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应用RT-PCR扩增中国小麦花叶病毒（CWMV）外壳蛋白通读区（readthrough,RT）5’端（RTn）和3’端（RTc）及19ku的富含半胱氨酸蛋白基因,分别克隆并在E.coli中表达,表达蛋白粗提纯后免疫小鼠,制备相应的抗血清和单克隆抗体.用各种抗体检测小麦病叶汁液中RTn,RTc和19ku蛋白.结果表明,RTn和RTc分布在病叶细胞中的CWMV病毒粒子表面,而19ku蛋白则均匀分布于细胞质中.
RT-PCR was used to amplify the 5 ’(RTn) and 3’ end (RTc) of the coat protein of Chinese wheat mosaic virus (CWMV) and the 19ku cysteine rich protein gene, And expressed in E.coli.The recombinant protein was purified and the mice were immunized.The antisera and monoclonal antibodies were prepared.The RTn, RTc and 19ku proteins in diseased leaves of wheat were detected by various antibodies.The results showed that RTn and RTc CWMV virus particles distributed in the diseased leaf surface, while the 19ku protein is evenly distributed in the cytoplasm.