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Yeast two-hybrid analysis of jasmonate signaling proteins
Amparo Pérez Cuéllar1, Laurens Pauwels, Rebecca De Clercq
1Department of Plant Systems Biology, VIB, Gent, Belgium.
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.
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