Ma-Högemeier Zhan-Lu, Zhu Qin-Chang, Zheng Chunfu
College of Pharmacy, Shenzhen Technology University, Guangdong, China.
Department of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, AB, Canada.
Methods Mol Biol. 2025;2940:187-195. doi: 10.1007/978-1-0716-4615-1_17.
Bimolecular fluorescence complementation (BiFC) is a powerful technique used to visualize protein-protein interactions (PPIs) in living cells. By reconstituting a fluorescent protein from two nonfluorescent fragments fused to proteins of interest, BiFC enables noninvasive observation of protein interactions within the cellular environment. This method has been successfully applied to study a variety of biological systems, including the interactions of viral proteins with host factors, thereby enhancing our understanding of viral pathogenesis and developing targeted therapeutic strategies. In this chapter, we outline the application procedure and provide troubleshooting notes for BiFC when conducted in virological studies.
双分子荧光互补(BiFC)是一种用于在活细胞中可视化蛋白质-蛋白质相互作用(PPI)的强大技术。通过从与感兴趣的蛋白质融合的两个无荧光片段重构荧光蛋白,BiFC能够在细胞环境中对蛋白质相互作用进行非侵入性观察。该方法已成功应用于研究各种生物系统,包括病毒蛋白与宿主因子的相互作用,从而增进了我们对病毒发病机制的理解并制定了靶向治疗策略。在本章中,我们概述了应用程序,并提供了在病毒学研究中进行BiFC时的故障排除说明。