Luciferase Complementation Assay for Protein‐Protein Interactions in Plants

Abstract Constitutive and dynamic protein‐protein interactions are fundamental to all aspects of cellular processes. Compared to other techniques measuring protein‐protein interactions in plants, the luciferase complementation assay has a number of advantages: it detects plant protein‐protein interactions in real time, requires little hands‐on manipulation of samples, is highly quantitative, has extremely low background, and can be easily scaled up for high‐throughput interactome studies. Here, we describe a protocol that includes two alternate data collection methods to quantify luminescence results based on Agrobacterium ‐mediated transient luciferase expression in Nicotiana benthamiana . One data collection method employs a charge‐coupled device imaging system that allows the interactions to be presented as images, and the other employs a luminometer, which enables the assay to be conducted in a 96‐well plate. Technical parameters related to frequently encountered problems and common errors, presented here, are important for performing this assay successfully. © 2018 by John Wiley & Sons, Inc.