| 中国小麦花叶病毒运动蛋白基因的克隆及其表达 |
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| 引用本文： | 沃恩康,郑滔,张巧艳,陈剑平.中国小麦花叶病毒运动蛋白基因的克隆及其表达[J].科技通报,2006,22(3):319-322. |
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| 作者姓名： | 沃恩康 郑滔 张巧艳 陈剑平 |
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| 作者单位： | 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 |
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| 基金项目： | 国家杰出青年科学基金;浙江省科技厅资助项目 |
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| 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。
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| 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 |
| 文章编号： | 1001-7119（2006）03-0319-04 |
| 收稿时间： | 2005-01-24 |
| 修稿时间： | 2005年1月24日 |
Cloning and Expression of Movement Protein Gene of Chinese Wheat Mosaic Virus (CWMV) |
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| WO En-kang,ZHENG Tao,ZHANG Qiao-yan,CHEN Jian-ping.Cloning and Expression of Movement Protein Gene of Chinese Wheat Mosaic Virus (CWMV)[J].Bulletin of Science and Technology,2006,22(3):319-322. |
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| Authors: | WO En-kang ZHENG Tao ZHANG Qiao-yan CHEN Jian-ping |
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| Institution: | 1. College of Life Science, Zhejiang University, Hangzhou 310029, China; 2. Institute of Virology and Biotechnology, Zhejiang Academy of Agriculture Sciences, Hangzhou 310021, China |
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| 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. |
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| Keywords: | molecular biology Chinese wheat mosaic virus movement protein gene expression yeast two-hybrid system |
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