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

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。

Abstract: 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.