云南省马铃薯番茄斑萎病毒N蛋白自身互作功能域鉴定

Identification of Self-Interacting Domains of the Nucleocapsid Protein of Tomato Spotted Wilt Virus of Potato in Yunnan Province

为了探究番茄斑萎病毒核衣壳蛋白N在马铃薯上的致病性,采用酵母双杂交和双分子荧光互补技术研究了病毒N蛋白的自身互作及其关键功能域。通过RT-PCR扩增了N基因全长及其截断突变体,并构建了酵母表达载体对其自身互作的功能区域进行鉴定。结果显示,在酵母细胞中N蛋白自身互作的关键功能域位于186位至193位氨基酸之间,且N蛋白在植物体内的互作主要发生在细胞核和细胞质中。研究结果可为番茄斑萎病毒的防控提供理论依据。

In order to explore the pathogenicity of nucleocapsid protein N of tomato spotted wilt virus in potato,the self-interaction and key functional domains of viral N protein were studied by yeast two-hybrid and bimolecular fluorescence complementation techniques.The full-length N gene and its truncated mutants were amplified by RT-PCR,and yeast expre-ssion vectors were constructed to identify the functional regions interacting with themselves.The results showed that the key functional domain of N protein self-interaction in yeast cells was located between amino acid 186 and amino acid 193,and the interaction of N protein in plants mainly occurred in the nucleus and cytoplasm.The research results can provide a theoretical basis for the prevention and control of the outbreak of tomato spotted wilt virus.