拟南芥ROP2与GGB蛋白互作的分析  

Analysis of Interaction between ROP2 and GGB Proteins of Arabidopsis thaliana

作　　者：杨迷 胡燕 李继洋[1] 李月[1] 刘超[1] 代培红[1] 刘晓东[1] Yang Mi;Hu Yan;Li Jiyang;Li Yue;Liu Chao;Dai Peihong;Liu Xiaodong(Laboratory of Agricultural Biotechnology of Xinjiang Agricultural University,College of Agronomy,Xinjiang Agricultural University,Urumqi,830052)

机构地区：[1]新疆农业大学农学院,新疆农业大学农业生物技术重点实验室,乌鲁木齐830052

出　　处：《分子植物育种》2020年第7期2216-2224,共9页Molecular Plant Breeding

基　　金：国家自然基金(31660433)资助。

摘　　要：为了阐明拟南芥ROP蛋白家族成员ROP2与蛋白异戊二烯化中I型香叶酰基的β亚基GGB的互作关系,本研究先后采用酵母双杂交和三杂交技术分析了两者之间的相互作用。结果表明,ROP2与GGB之间在酵母细胞中并没有发生相互作用。我们又采用双分子荧光互补(BiFC)技术在植物体内再次验证两者之间的相互作用。重新构建了一套带多克隆位点的常规BiFC重组表达载体,基于这套BiFC载体,进行了验证分析。结果表明,实验组在本氏烟草的表皮细胞中发现了黄色荧光信号,并且信号定位在细胞膜上,而阴性对照组未发现荧光信号,说明GGB蛋白与ROP2蛋白在植物体内存在相互作用,并且这种相互作用发生在细胞膜上。本研究结果为进一步揭示ROP2的生化调控机制奠定了前期基础。In order to elucidate the interaction between ROP2 of ROP protein family andβsubunit GGB of type I geranyl group in protein is prenylation in Arabidopsis thaliana,yeast two-hybrid and three-hybrid techniques were used to analyze the interaction between them.The results showed that there was no interaction between ROP2 and GGB in yeast cells.In addition,we used the bimolecular fluorescence complementation(BiFC)technology to verify the interaction between ROP2 and GGB in plants.Firstly,a set of conventional BiFC recombinant expression vectors with multiple cloning sites was reconstructed and used to validate.The experimental results showed that yellow fluorescent signals were found in the epidermal cells of tobacco in the experimental group and located on the cell membrane,while no fluorescent signals were found in the negative control group,indicating that there was interaction between GGB and ROP2 in plants and the interaction occured in cell membrane.This study laid a foundation for further revealing the regulation mechanism of ROP2.

关 键 词：酵母杂交 双分子荧光互补(BiFC) GGB ROP2 蛋白质互作 酶切连接 

分 类 号：Q943.2[生物学—植物学]