拟南芥膜蛋白与AtGSL5互作筛选研究  

Screening for Arabidopsis Membrane Proteins Interacting with AtGSL5

作　　者：秦宏坤 李明卓 秦冰倩 胡云丹 李敏[1,2] QIN Hongkun;LI Mingzhuo;QIN Bingqian;HU Yundan;LI Min(School of Life Science,Gannan Normal University;National Navel Orange Engineering Research Center,Ganzhou 341000,China)

机构地区：[1]赣南师范大学生命科学学院 [2]国家脐橙工程技术研究中心,江西赣州341000

出　　处：《赣南师范大学学报》2024年第6期84-90,共7页Journal of Gannan Normal University

基　　金：国家自然科学基金青年项目(32202272)。

摘　　要：通过构建拟南芥膜蛋白酵母双杂交cDNA文库,筛选ATGSL5互作蛋白,为探究压力胁迫下植物调控胼胝质合成的信号途径提供参考.通过提取整株拟南芥RNA,分离出mRNA.以mRNA为模板,利用SMART技术成功合成双链cDNA,将双链cDNA和pPR3-N载体用T4 DNA连接酶进行连接.连接产物电转化到HST08感受态细胞,构建拟南芥膜蛋白酵母双杂交cDNA文库并进行质量检测.结果表明,总库容量达到3×10^(7)cfu,重组率为100%,符合膜蛋白酵母双杂交实验要求.利用该文库筛选到了ATGSL5互作蛋白AT5G43460.The split-ubiquitin membrane-based yeast two-hybrid system presented in this study was used to construct a yeast two-hybrid cDNA library of Arabidopsis thaliana.The whole RNA of Arabidopsis thaliana was extracted,and the mRNA was purified.cDNA is generated from mRNA using SMART technology.The double trand cDNA and the pPR3-N vector were ligated with T4 DNA ligase.The recombinant vectors were electrotransformed into HST08 competent cells to construct a membrane protein yeast two-hybrid cDNA library.The quality of the library was then identified.The capacity of the library was 3×10^(7) cfu and the recombination rate was 100%.The AtGSL5 interacting protein AT5G43460 was screened using this library.

关 键 词：膜蛋白酵母双杂交 CDNA文库 拟南芥 蛋白质相互作用 

分 类 号：Q71[生物学—分子生物学]