小麦翻译控制肿瘤蛋白(TCTP)与索马甜类蛋白(TLP)的相互作用

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摘要摘要翻译控制肿瘤蛋白(translationally controlled tumor protein, TCTP)分布于真核细胞中，参与包括有丝分裂、DNA损伤修复和植物抵御病原物侵染等多种生物进程。索马甜类蛋白(thaumatin-like protein, TLP)是一类植物中的病程相关蛋白，参与多种植物响应病原物侵染的过程。本实验室前期研究证明TaTCTP参与小麦(Triticum aestivum)抵御小麦叶锈菌侵染(Puccinia triticina)的防卫反应。本研究关注小麦TaTCTP和TaTLP之间的相互作用，分别构建了用于酵母双杂交(yeast two- hybrid)和双分子荧光互补(Bi-molecular fluorescence complementation, BiFC)实验的载体。通过酵母双杂交实验，发现同时携带表达TCTP和TLP载体的酵母AH109可以在SD-Leu/-Trp/-His和SD-Leu/-Trp/-His/-Ade培养基上生长，并可检测到报告基因α-半乳糖苷酶基因(α- galactosidase gene, MEL1)活性，表明TaTCTP和TaTLP可发生物理互作。通过BiFC实验，发现共同转化表达TCTP和TLP载体的烟草(Nicotiana benthamiana)下表皮细胞的细胞质中可观察到强烈的黄色荧光，表明TaTCTP和TaTLP可在细胞质中发生相互作用。本研究为深入研究TCTP在小麦抵抗小麦叶锈菌侵染中的作用机制奠定基础。

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	麻楠
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	王冬梅

关键词 ：翻译控制肿瘤蛋白(TCTP), 索马甜类蛋白(TLP), 小麦, 酵母双杂交, 双分子荧光互补

Abstract：Abstract Translationally controlled tumor protein (TCTP) exists widely in eukaryotic cells. TCTP was involved in a variety of cellular processes including mitotic regulation, DNA damage repair, and plant resistance to pathogen infection. Thaumatin-like protein (TLP) is a pathogenesis related protein in plants and is thought to participate in the various processes of plants in response to pathogens infection. Previous work in this laboratory has demonstrated that TaTCTP was involved in the response of wheat (Triticum aestivum) to Puccinia triticina infection. This study focused on the interaction between TCTP and TLP. Vectors were constructed for yeast two-hybrid and Bi-molecular fluorescence complementation (BiFC), respectively. Through yeast two-hybrid test, it was found that yeast AH109 carried both TCTP and TLP could grow on SD-Leu/-Trp/-His and SD-Leu/-Trp/-His/-Ade medium and have detectable α- galactosidase gene (MEL1) reporter activity. This result indicated that there is a physical interaction between TaTCTP and TaTLP. Through the BiFC test, it was found that a strong yellow fluorescence signal could be observed in tobacco epidermis cytoplasm when co-transforming Agrobacterium with TCTP and TLP. These results indicated that TaTCTP and TaTLP can interact with each other in cytoplasm. This study lay a foundation for further exploring the function and mechanism of TCTP in wheat resistance to Puccinia triticina infection.

Key words： Translationally controlled tumor protei (TCTP) Thaumatin-like protein (TLP) Wheat Yeast two- hybrid Bi-molecular fluorescence complementation (BiFC)

收稿日期: 2017-11-08 出版日期: 2018-05-21

基金资助:国家自然科学基金;高等学校博士学科点专项科研基金资助课题（优先发展领域）;河北省应用基础研究计划重点基础研究项目

通讯作者: 王冬梅 E-mail: dongmeiwang63@126.com

引用本文:

麻楠乔金柱孙天杰李姗魏凤菊王冬梅. 小麦翻译控制肿瘤蛋白(TCTP)与索马甜类蛋白(TLP)的相互作用[J]. , 2018, 26(6): 911-919.

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http://journal05.magtech.org.cn/Jwk_ny/CN/ 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2018/V26/I6/911

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