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用于检测植物转录因子与其他调控蛋白相互作用的原位邻近连接分析

In Situ Proximity Ligation Assay to Detect the Interaction Between Plant Transcription Factors and Other Regulatory Proteins.

作者信息

Xu Yifeng, Gan Eng-Seng, Ito Toshiro

机构信息

Temasek Life Sciences Laboratory (TLL), 1 Research Link, National University of Singapore, Singapore, Singapore.

Biological Sciences, Nara Institute of Science and Technology, Ikoma, Nara, Japan.

出版信息

Methods Mol Biol. 2018;1830:325-335. doi: 10.1007/978-1-4939-8657-6_19.

Abstract

In plants, a lot of transcription factors fulfill their roles in gene regulation through the interaction with other regulatory proteins and co-factors. Thus, confirmation of protein-protein interaction is key to understand the precise function of transcription factors. Many methods have been developed to investigate the protein-protein interaction in vivo and in vitro. In situ Proximity Ligation Assay (PLA) is an innovative method to test protein-protein interaction in your tissues or cells of interest in vivo. Furthermore, this method allows us to detect transient interaction and low-abundance protein interaction with single molecule resolution. In this chapter, we describe a detailed protocol for the study of interaction between plant transcription factors and other regulatory proteins, in the scale of single nuclei of plant organ, tissues and cells.

摘要

在植物中,许多转录因子通过与其他调节蛋白和辅助因子相互作用来履行其在基因调控中的作用。因此,证实蛋白质-蛋白质相互作用是理解转录因子精确功能的关键。已经开发了许多方法来研究体内和体外的蛋白质-蛋白质相互作用。原位邻近连接分析(PLA)是一种在体内你感兴趣的组织或细胞中检测蛋白质-蛋白质相互作用的创新方法。此外,该方法使我们能够以单分子分辨率检测瞬时相互作用和低丰度蛋白质相互作用。在本章中,我们描述了一个详细的方案,用于在植物器官、组织和细胞的单核水平上研究植物转录因子与其他调节蛋白之间的相互作用。

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