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Updated: May 12, 2026

Luciferase Complementation Imaging Assay in Nicotiana benthamiana Leaves for Transiently Determining Protein-protein Interaction Dynamics
Published on: November 20, 2017
Pull-down analysis of interactions among jasmonic acid core signaling proteins
Sandra Fonseca1, Roberto Solano
1Departamento de Genética Molecular de Plantas, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
Abstract:
Pull-down assays are key tools to test specific protein-protein interactions and have been particularly fruitful in jasmonate signaling research. Here, we describe a standard protocol in which a matrix-bound "bait" protein, expressed in Escherichia coli, pulls down a "prey" protein that is soluble in a protein extract obtained from Arabidopsis thaliana plant tissues. The pulled-down protein can be detected by immunoblot with protein-specific or epitope-specific antibodies. Such a pull-down method was used to reveal interactions among components of the jasmonic acid signaling, including hormone-dependent coreceptor interaction, homodimerization and heterodimerization of JASMONATE ZIM DOMAIN repressors, and interactions among other corepressor components and with transcription factors. Pull-down assays contributed not only to shape this signaling pathway but also to identify the active jasmonate hormone.
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