Institute of Plant Stress Biology, State Key Laboratory of Cotton Biology, School of Life Sciences, Henan University, Kaifeng, China.
College of Biological Sciences, Hebei Normal University, Shijiazhuang, Hebei, China.
Methods Mol Biol. 2022;2398:205-213. doi: 10.1007/978-1-0716-1912-4_16.
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.
萤火虫荧光素酶互补分析（FLCA）是分析组成型和动态蛋白质-蛋白质相互作用（PPIs）最广泛使用的灵敏和可靠的方法之一。在这里，我们报告了一种通过 Topcount™微孔板闪烁计数器或深冷 CCD 相机在拟南芥 F1 杂种中进行长期体内检测植物蛋白-蛋白相互作用的方法。按照这些方案，我们成功地检测了 EARLY FLOWERING 3（ELF3）和 EARLY FLOWERING 4（ELF4）的时间依赖性 PPIs；它们都与 LUX ARRHYTHMO（LUX）一起属于一个夜间复合物，该复合物已被发现在昼夜节律、开花和生长中起关键作用。
