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草业科学  2014 

林烟草8个防卫反应信号蛋白编码基因的克隆及RNAi植物表达载体的构建

DOI: 10.11829\j.issn.1001-0629.2013-0195, PP. 1275-1282

Keywords: 林烟草,烟草花叶病毒,RNA干扰

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Abstract:

?为了分析林烟草(Nicotianasylvestris)N’基因对烟草花叶病毒(Tobaccomosaicvirus,TMV)产生抗性的信号途径,培育抗TMV植物新品种,选取PAD4、EDS1、Rar1、Sgt1、NPR1、Jar6、COI1、CTR1作为候选基因。以林烟草总RNA为模板,通过反转录分别克隆上述8个基因的片段,通过添加酶切位点以及酶切、连接和转化,成功构建了上述8个信号蛋白的RNAi载体。酶切验证结果表明,所构建的8个RNAi载体的结构正确,可以用于进一步的遗传转化试验,这为分析N’基因对TMV产生抗性所依赖的信号途径奠定了基础。

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