马铃薯StMAPKK1互作蛋白的酵母双杂交筛选及生物信息学分析

Yu-Huang Liao, Fangfang Wang, Xi Zhu, Ning Zhang, H. Si
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摘要

丝裂原活化蛋白激酶(Mitogen-activated protein kinase, MAPK)级联反应是参与植物生长发育、激素和胁迫反应的重要而复杂的信号网络之一。作为其主要成员之一,丝裂原活化蛋白激酶(MAPKK)位于级联反应的中间,在信号的收集和发散中起着关键作用。有研究表明马铃薯(Solanum tuberosum L.)StMAPKK1 (PGSC0003DMT400000744)基因响应干旱胁迫。因此,首先选择StMAPKK1基因筛选其相互作用蛋白。本研究通过同源重组构建诱饵载体pGBKT7-StMAPKK1,利用酵母双杂交系统对马铃薯cDNA文库进行筛选。筛选得到5个与stmapkk1相互作用的蛋白,分别是水解o -糖基化合物(hydrolase)、RING-H2亚群RHE蛋白、氰酸酯水合酶、ARF GTPase激活剂和一个C2结构域蛋白,通过生物信息学分析进行鉴定,并通过小规模杂交验证相互作用。该结果为进一步研究马铃薯StMAPKK1的信号通路和生物学功能提供了理论依据。
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Screening and Bioinformatics Analysis of Proteins Interacting with StMAPKK1 in Potato by Yeast Two-Hybrid System
Mitogen-activated protein kinase (MAPK) cascade reaction is one of important and complex signal networks involved in plant growth and development, hormone and stress response. As one of its main members, mitogen-activated protein kinase kinase (MAPKK) is located in the middle of the cascade reaction and plays a key role in signal collection and divergence. There were shown that potato ( Solanum tuberosum L.) StMAPKK1 (PGSC0003DMT400000744) gene responded to drought stress. Therefore, StMAPKK1 gene was firstly selected to screen its interacting protein. In this study, the bait vector pGBKT7-StMAPKK1 was constructed by homologous recombination and used to screen potato cDNA library by yeast ( Saccharomyces cerevisiae ) two-hybrid system. Five StMAPKK1-interacting proteins, hydrolase (hydrolyzing O-glycosyl compound), RING-H2 subgroup RHE protein, cyanate hydratase, ARF GTPase activator, and a C2 domain-containing protein, were obtained through this screening and were identificated by bioinformatics analysis, and the interaction was verified by small-scale hybridization verification. The results provided theoretical basis for further study on the signal pathway and biological function of potato StMAPKK1.
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