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.
CITATION STYLE
Singh, R., Dangol, S., & Jwa, N. S. (2014). Yeast two-hybrid system for dissecting the rice MAPK interactome. Methods in Molecular Biology, 1171, 195–216. https://doi.org/10.1007/978-1-4939-0922-3_16
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