title: Yeast two-hybrid analysis of jasmonate signaling proteins.
abstract: Protein-protein interaction studies are crucial to unravel how jasmonate (JA) signals are transduced. Among the different techniques available, yeast two-hybrid (Y2H) is commonly used within the JA research community to identify proteins belonging to the core JA signaling module. The technique is based on the reconstitution of a transcriptional activator that drives the reporter gene expression upon protein-protein interactions. The method is sensitive and straightforward and can be adapted for different approaches. In this chapter, we provide a detailed protocol to perform targeted Y2H assays to test known proteins and/or protein domains for direct interaction in a pairwise manner and present the possibility to study ternary protein complexes through Y3H.
authors.affiliation: Department of Plant Systems Biology, VIB, Gent, Belgium.
authors.forename: Amparo Pérez
authors.initials: AP
authors.lastname: Cuéllar
authors.forename: Laurens
authors.initials: L
authors.lastname: Pauwels
authors.forename: Rebecca
authors.initials: R
authors.lastname: De Clercq
authors.forename: Alain
authors.initials: A
authors.lastname: Goossens
mesh_terms: D017360:Arabidopsis; D029681:Arabidopsis Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D004926:Escherichia coli; D005822:Genetic Vectors; D047908:Intracellular Signaling Peptides and Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D011485:Protein Binding; D012441:Saccharomyces cerevisiae; D014170:Transformation, Genetic; D020798:Two-Hybrid System Techniques
chemical_list: D029681:Arabidopsis Proteins; D047908:Intracellular Signaling Peptides and Proteins