Pham Cau D
Department of Pharmacology, Emory University School of Medicine, Atlanta, GA, USA,
Methods Mol Biol. 2015;1278:483-95. doi: 10.1007/978-1-4939-2425-7_32.
The bimolecular fluorescence complementation (BiFC) assay is a versatile technique for investigating protein-protein interaction (PPI) in living systems. The BiFC assay exploits the color-emitting moiety and the modular structure of fluorescent proteins to provide both temporal and spatial information of the PPI. The modular property of fluorescent proteins enables researchers to strategically partition a fluorescent protein into two nonfluorescent units, which can be independently fused to other proteins. When the fusion proteins interact with each other, the nonfluorescent fragments reconstitute to generate a fluorescence signal. PPI can then be detected by capturing the fluorescence signal with a fluorescence microscope. In this chapter, the Venus fluorescent protein is employed to demonstrate the application of the BiFC assay.
双分子荧光互补（BiFC）分析是一种用于研究活细胞系统中蛋白质-蛋白质相互作用（PPI）的通用技术。BiFC分析利用荧光蛋白的发色基团和模块化结构来提供PPI的时间和空间信息。荧光蛋白的模块化特性使研究人员能够将荧光蛋白策略性地分割成两个无荧光的单元，这两个单元可以独立地与其他蛋白质融合。当融合蛋白相互作用时，无荧光的片段重新组装以产生荧光信号。然后可以通过荧光显微镜捕获荧光信号来检测PPI。在本章中，使用维纳斯荧光蛋白来演示BiFC分析的应用。
