Chinese Bulletin of Botany ›› 2020, Vol. 55 ›› Issue (1): 62-68.DOI: 10.11983/CBB19143

• TECHNIQUES AND METHODS • Previous Articles     Next Articles

Pull-down and Co-immunoprecipitation Assays of Interacting Proteins in Plants

Chongyi Xu()   

  1. Key Laboratory of Plant Molecular Physiology, CAS Center for Excellence in Molecular Plant Sciences, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China
  • Received:2019-08-01 Accepted:2019-08-30 Online:2020-01-01 Published:2019-12-20
  • Contact: Chongyi Xu

Abstract: Protein-protein interactions play a key role in cellular signaling, involved in various biological processes. Studies on these interactions are therefore crucial toward understanding the regulatory networks of cellular signaling. It is a standard practice that the protein-protein interactions identified by the yeast two-hybrid system should be independently confirmed by in vitro and in vivo approaches. Pull-down and co-immunoprecipitation (Co-IP) are routine approaches to detect protein-protein interactions. Pull-down assay is used to detect direct or physical interactions between proteins in vitro. In plant biology studies, one of the most convenient methods to detect protein-protein interactions is the transient expression of the target proteins in Nicotiana benthamiana leaves followed by the Co-IP assay. In this paper, we describe the principles and protocols for the GST tag-based pull-down assay and the Co-IP assay of proteins transiently expressed in N. benthamiana leaves, providing a reference for detecting plant protein-protein interactions.

Key words: plants, protein-protein interactions, Pull-down, Co-immunoprecipitation