Firefly Luciferase Complementation-Based Analysis of Dynamic Protein-Protein Interactions Under Diurnal and Circadian Conditions in Arabidopsis

Methods Mol Biol. 2022:2398:205-213. doi: 10.1007/978-1-0716-1912-4_16.

Qiguang Xie ¹ ², Qiao Wang ³, Xiaodong Xu ³ ⁴

Affiliations

1 Institute of Plant Stress Biology, State Key Laboratory of Cotton Biology, School of Life Sciences, Henan University, Kaifeng, China. qiguang.xie@vip.henu.edu.cn.
2 College of Biological Sciences, Hebei Normal University, Shijiazhuang, Hebei, China. qiguang.xie@vip.henu.edu.cn.
3 Institute of Plant Stress Biology, State Key Laboratory of Cotton Biology, School of Life Sciences, Henan University, Kaifeng, China.
4 College of Biological Sciences, Hebei Normal University, Shijiazhuang, Hebei, China.

PMID: 34674178 DOI: 10.1007/978-1-0716-1912-4_16

Abstract

Split firefly luciferase complementation assay (FLCA) is one of the most widely used sensitive and reliable methods for the analysis of constitutive and dynamic protein-protein interactions (PPIs). Here, we report a method for long-term in vivo detects plant protein-protein interactions in Arabidopsis F1 hybrids via Topcount™ Microplate Scintillation Counter or Deep-Cooled CCD camera. Following these protocols, we successfully detected time-dependent PPIs of EARLY FLOWERING 3 (ELF3) and EARLY FLOWERING 4 (ELF4); both of them with LUX ARRHYTHMO (LUX) belong to an evening complex which has been found to play a key role in circadian rhythms, flowering, and growth.

Keywords

Bioluminescence; Circadian rhythm; Diurnal rhythm; Dynamic protein-protein interactions; Firefly luciferase complementation.

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