利用酵母双杂交筛选木薯果胶甲酯酶抑制因子MePMEI1的互作蛋白  

Screening of Pectin Methylesterase Inhibitor Me PMEI1-interactive Proteins in Cassava(Manihot esculenta) Using Yeast Two-hybrid System

作　　者：仇婷婷 李瑞梅[2,3] 周杨骄 林雪君 姚远[2,3] 刘姣[2,3] 符少萍[2,3] 胡新文 郭建春[2,3] Qiu Tingting;Li Ruimei;Zhou Yangjiao;Lin Xuejun;Yao Yuan;Liu Jiao;Fu Shaoping;Hu Xinwen;Guo Jianchun(College of Tropical Agriculture and Forestry,Hainan University,Haikou,570228;Institute of Tropical Bioscicnce and Biotechnology,Chinese Acadcmy of Tropical Agriculture Sciences,Haikou,571101;Key Laboratory of Ministry of Agriculture for Biology and Genetic Resources of Tropical Crops,Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Sciences,Haikou,571101)

机构地区：[1]海南大学热带农林学院,海口570228 [2]中国热带农业科学院热带生物技术研究所,海口571101 [3]中国热带农业科学院热带生物技术研究所,农业部热带作物生物学与遗传资源利用重点实验室,海口571101

出　　处：《基因组学与应用生物学》2021年第4期1626-1633,共8页Genomics and Applied Biology

基　　金：国家自然科学基金项目(31601359,31600196,31671767);现代农业产业技术体系建设专项(CARS-11-HNGJC);国家重点研发计划项目(2018YFD1000500);海南省重点研发计划项目(ZDYF20-17073);海南省2018年研究生创新科研课题(Hys2018-03)共同资助。

摘　　要：果胶甲酯酶抑制因子(PMEI)在植物抗逆过程中具有重要作用。为了探究木薯MePMEI1的生物学功能及其在蛋白互作网络中的作用,本研究通过构建诱饵载体pGBKT7-MePMEI1,通过酵母双杂交方法从木薯cDNA文库中筛选Me PMEI1的互作蛋白。结果表明,酵母双杂交共筛选出57个阳性克隆,阳性率约为32.6%,测序鉴定出37种含完整开放阅读框(ORF)的候选互作蛋白,包括非特异性丝氨酸/苏氨酸蛋白激酶/苏氨酸特异性蛋白激酶、叶绿素a/b结合蛋白、电压依赖性阴离子通道蛋白(VDAC2)、乙二醛酶Glyoxalase-2、氮素调节蛋白、木糖葡萄糖转移酶(TCH4)等,并通过回转验证检验其蛋白互作的真实性。验证结果表明Me PMEI1与VDAC2、Glyoxalase-2蛋白具有相互作用。这进一步补充了其蛋白互作网络,为探究MePMEI1蛋白的互作网络提供新的依据。Pectin methylesterase inhibitor(PMEI) plays an important role in the process of plant stress resistance.In order to explore the biological function of MePMEI1 in cassava and its role in protein interaction network, in this paper, the bait vector pGBKT7-MePMEI1 was constructed in cassava and the interaction proteins of MePMEI1 were screened in cassava c DNA library by yeast two-hybrid. The results showed that a total of 57 positive clones were screened, and the positive rate was about 32.6%. About 37 kinds candidate interaction proteins with complete open reading frame(ORF) were identified by sequencing, including non-specific serine/threonine protein kinase, threonine specific protein kinase, chlorophyll a/b binding protein, voltage dependent anion channel protein2(VDAC2), nitrogen regulatory protein, xyloseglucose transferase(TCH4), Glyoxalase-2 protein, etc. The authenticity of protein interaction was verified by rotary verification. The results showed that MePMEI1 interacted with VDAC2, glyoxalase-2 protein, which further supplemented its protein interaction network, and provided new evidence for exploring the interaction network of MePMEI1 protein.

关 键 词：木薯 果胶甲酯酶抑制因子 酵母双杂交 电压依赖性阴离子通道蛋白 

分 类 号：S533[农业科学—作物学]