【摘要】 应用RT-PCR技术从小麦病叶中扩增得到中国小麦花叶病毒(CWMV)的运动蛋白(MP)基因,在大肠杆菌BL21(DE3)plysS中得到大量表达,并以含表达产物的凝胶为抗原,免疫小鼠制备特异性抗血清,同时将MP基因重组到质粒pGBKT7中,电击转化酵母Y187菌株。Westernblot分析表明MP基因在酵母体内能以融合蛋白的形式正确表达,并具有免疫活性,可作为酵母双杂交系统中的“诱饵”质粒。更多还原

【Abstract】 The movement protein(MP)gene of Chinese wheat mosaic virus(CWMV)was amplified from infected wheat leaves by RT-PCR and was highly expressed in E.coli BL21(DE3)plysS.The MP was purified by SDS-PAGE and the antiserum against the protein was raised in mouse.The recombinant plasmid pGBKT7-MP was constructed by ligating the gene of MP into pGBKT7,then transformed into yeast Y187 to abserve MP expression by western blot.As a result,the MP was expressed as recombinant protein and has immunogenic activity.pGBKT7-MP can serve as a bait plasmid of yeast two-hybrid system.更多还原