Development of a reverse-genetics system for Tomato spotted wilt virus

番茄斑萎病毒反向遗传学系统的开发

• 批准号: 23658043
• 负责人: ISHIKAWA Masayuki
• 金额: $ 2.58万
• 依托单位: National Institute of Agrobiological Sciences
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-23658043/
• 关键词: 感染・増殖; 植物; マイナス鎖RNAウイルス; 複製; 転写; 宿主因子

Toward establishing an experimental system in which infection with Tomato spotted wilt virus(TSWV) can be initiated from its cDNA, we expressed TSWV nucleocapsid protein and RNA polymerase(the L protein) in Escherichia coli and Saccharomyces cerevisiae, respectively. We purified these proteins and mixed with in vitro-synthesized TSWV RNA in various conditions, however, we could not detect RNA synthesis. We also constructed plasmids to express TSWV-coded proteins and RNA in vivo, and introduced these plasmids into Nicotiana benthamiana leaves by the Agro-infiltration method, infection with TSWV did not occur. In the course of these trials, we obtained results that suggest that the translation elongation factor eEF1A plays important roles in mRNA transcription of TSWV.

为了建立番茄斑萎病毒（Tomato spotted wilt virus，TSWV）感染的实验体系，我们分别在大肠杆菌和酿酒酵母中表达了TSWV核衣壳蛋白和RNA聚合酶（L蛋白）。我们纯化了这些蛋白质，并在各种条件下与体外合成的TSWV RNA混合，然而，我们不能检测到RNA合成。我们还构建了TSWV编码蛋白和RNA的体内表达质粒，并通过农杆菌渗入法将这些质粒导入本氏烟草叶片中，没有发生TSWV的感染。在这些试验的过程中，我们得到的结果表明，翻译延伸因子eEF 1A在TSWV的mRNA转录中起着重要作用。

项目成果

トマト黄化えそウイルスのmRNA合成を活性化する宿主因子の探索

寻找激活番茄黄化病毒mRNA合成的宿主因子

• 发表时间: 2012
• 作者: 薦田圭介;石橋和大;河村和恵;石川雅之;飯哲夫
• 通讯作者: 飯哲夫