Singh Raksha, Dangol Sarmina, Jwa Nam-Soo
Department of Molecular Biology, College of Life Sciences, Sejong University, 98 Gunja-dong, Gwangjin-gu, Seoul, 143-747, Republic of Korea.
Methods Mol Biol. 2014;1171:195-216. doi: 10.1007/978-1-4939-0922-3_16.
Protein-protein interactions are a preliminary but fundamental key to many biological systems. Identification of proteins that interact with particular bait not only contributes to a deeper understanding of bait protein function but also provides much information for the discovery of larger-scale interaction networks (interactome). Therefore, protein-protein interaction mapping is regarded as a widely accepted standardized functional genomics technique that provides comprehensive functional interpretation of previously uncharacterized proteins. A commonly used approach to detecting novel protein-protein interactions is the yeast two-hybrid system. In this chapter we describe in detail the protocols used to dissect the rice MAPK interactome, including the bait protein auto-activation test, identification of a rice MAPK interacting protein, confirmation of interaction by retransformation assay and characterization of the novel interacting protein.
蛋白质-蛋白质相互作用是许多生物系统的一个初步但基本的关键。鉴定与特定诱饵相互作用的蛋白质不仅有助于更深入地了解诱饵蛋白质的功能，还为发现更大规模的相互作用网络（相互作用组）提供了大量信息。因此，蛋白质-蛋白质相互作用图谱绘制被视为一种广泛接受的标准化功能基因组学技术，它能对以前未表征的蛋白质进行全面的功能解释。检测新型蛋白质-蛋白质相互作用的常用方法是酵母双杂交系统。在本章中，我们详细描述了用于剖析水稻丝裂原活化蛋白激酶（MAPK）相互作用组的实验方案，包括诱饵蛋白自激活测试、水稻MAPK相互作用蛋白的鉴定、通过再转化试验确认相互作用以及新型相互作用蛋白的表征。
