中国小麦花叶病毒运动蛋白基因的克隆及其表达  

Cloning and Expression of Movement Protein Gene of Chinese Wheat Mosaic Virus(CWMV)

作　　者：沃恩康[1] 郑滔[2] 张巧艳[2] 陈剑平[2] 

机构地区：[1]浙江大学生命科学学院,杭州310029 [2]浙江省农业科学院病毒学与生物技术研究所,杭州310021

出　　处：《科技通报》2006年第3期319-322,共4页Bulletin of Science and Technology

基　　金：国家杰出青年科学基金(30225031);浙江省科技厅重大科技项目(2003C12023)

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。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 pGBKTT, 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, pGBKTT-MP can serve as a bait plasmid of yeast two-hybrid system.

关 键 词：分子生物学 中国小麦花叶病毒 运动蛋白 基因表达 酵母双杂交系统 

分 类 号：Q785[生物学—分子生物学] Q786