title: Yeast two-hybrid system for dissecting the rice MAPK interactome.
abstract: 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.
authors.affiliation: Department of Molecular Biology, College of Life Sciences, Sejong University, 98 Gunja-dong, Gwangjin-gu, Seoul, 143-747, Republic of Korea.
authors.forename: Raksha
authors.initials: R
authors.lastname: Singh
authors.forename: Sarmina
authors.initials: S
authors.lastname: Dangol
authors.forename: Nam-Soo
authors.initials: NS
authors.lastname: Jwa
mesh_terms: D004926:Escherichia coli / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics; D006836:Hydro-Lyases / Q000235:genetics; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D012275:Oryza / Q000201:enzymology / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D025941:Protein Interaction Mapping / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D014170:Transformation, Genetic; D020798:Two-Hybrid System Techniques; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D029701:Saccharomyces cerevisiae Proteins; C554167:URA3 protein, S cerevisiae; D020928:Mitogen-Activated Protein Kinases; D001616:beta-Galactosidase; D006836:Hydro-Lyases; C586384:HIS3 protein, S cerevisiae