利用TurboID邻近蛋白标记技术获得水稻互作蛋白组的实验方法  

Experimental method for obtaining interaction proteome using TurboID-based proximity labeling technology in rice

作　　者：刘佳欣 何明良 刘颖湘 卜庆云[2] 李秀峰[2] 王臻昱[2] 刘剑锋[1] 田晓杰[2] LIU Jiaxin;HE Mingliang;LIU Yingxiang;BU Qingyun;LI Xiufeng;WANG Zhenyu;LIU Jianfeng;TIAN Xiaojie(College of Life Sciences,Jilin Normal University,Siping 136000,China;Northeast Institute of Geography and Agroecology,Chinese Academy of Sciences,Harbin 150081,China;University of Chinese Academy of Sciences,Beijing 100049,China)

机构地区：[1]吉林师范大学生命科学学院,吉林四平136000 [2]中国科学院东北地理与农业生态研究所,黑龙江哈尔滨150081 [3]中国科学院大学,北京100049

出　　处：《土壤与作物》2023年第3期256-263,共8页Soils and Crops

基　　金：黑龙江省自然科学基金(YQ2022C038);中国科学院青年创新促进会项目(2021229);中国科学院东北地理与农业生态研究所青年科学家小组项目(2023QNXZ02).

摘　　要：TurboID邻近蛋白标记(TurboID-based proximity labeling,TbPL)技术是将诱饵蛋白融合TurboID生物素连接酶,在该工具酶的作用下将生物素标记到邻近蛋白上,最终通过链酶亲和素偶连磁珠将邻近蛋白富集下来,通过质谱分析的方式获得互作蛋白组。本文主要以水稻重要转录因子OsWRKY53为例,详细描述了利用TbPL技术获得水稻互作蛋白组的方法和注意事项。The TurboID-based proximity labeling(TbPL)technology is to fuse the bait protein with the TurboID biotin ligase;then mark the biotin to the neighboring protein of bait protein under the action of the tool enzyme;finally,enrich the neighboring protein through the streptavidin magnetic beads to obtain the interaction proteome through mass spectrometry analysis.Taking OsWRKY53,an important transcription factor,as an example,this article mainly describes the method and precautions for obtaining interaction proteome using TbPL technology in rice.

关 键 词：水稻 TurboID 邻近蛋白标记 互作蛋白组 实验方法 

分 类 号：Q756[生物学—分子生物学] S511[农业科学—作物学]