Weber-Boyvat Marion, Li Shiqian, Skarp Kari-Pekka, Olkkonen Vesa M, Yan Daoguang, Jäntti Jussi
Research Program in Cell and Molecular Biology, Institute of Biotechnology, University of Helsinki, 00014, Helsinki, Finland.
Methods Mol Biol. 2015;1270:277-88. doi: 10.1007/978-1-4939-2309-0_20.
Visualization of protein-protein interactions in vivo offers a powerful tool to resolve spatial and temporal aspects of cellular functions. The bimolecular fluorescence complementation (BiFC) makes use of nonfluorescent fragments of green fluorescent protein or its variants that are added as "tags" to target proteins under study. Only upon target protein interaction is a fluorescent protein complex assembled, and the site of interaction can be monitored by microscopy. In this chapter, we describe the method and tools for the use of BiFC in the yeast Saccharomyces cerevisiae and in mammalian cells.
在体内可视化蛋白质-蛋白质相互作用为解析细胞功能的空间和时间方面提供了一个强大的工具。双分子荧光互补(BiFC)利用绿色荧光蛋白或其变体的非荧光片段,将其作为“标签”添加到正在研究的靶蛋白上。只有在靶蛋白相互作用时,才会组装成荧光蛋白复合物,并且可以通过显微镜监测相互作用位点。在本章中,我们描述了在酿酒酵母和哺乳动物细胞中使用BiFC的方法和工具。