Ma-Högemeier Zhan-Lu, Qiu Hui-Ling, Zheng Chunfu
College of Pharmacy, Shenzhen Technology University, Shenzhen, Guangdong, China.
Department of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, AB, Canada.
Methods Mol Biol. 2025;2940:387-398. doi: 10.1007/978-1-0716-4615-1_34.
Fluorescence resonance energy transfer (FRET) operates on the principle of nonradiative energy transfer between a donor and an acceptor fluorophore when they are in close proximity (typically 1-10 nm). It is a valuable method for identifying protein-protein interactions in live cells, where not only the physiological condition but also the exact spatial information where interactions occur can be obtained. FRET offers significant advantages for studying molecular interactions and dynamics during viral infections, including high sensitivity and specificity, real-time monitoring capabilities, and versatility in experimental design. In this chapter, we outline the application procedure and provide troubleshooting notes for fluorescent protein-based FRET analysis, including single-molecule FRET (smFRET), when conducted in virological studies.
荧光共振能量转移（FRET）基于供体荧光团和受体荧光团在紧密接近时（通常为1-10纳米）的非辐射能量转移原理运行。它是一种用于识别活细胞中蛋白质-蛋白质相互作用的有价值方法，通过该方法不仅可以获得生理条件，还能得到相互作用发生的确切空间信息。FRET在研究病毒感染期间的分子相互作用和动力学方面具有显著优势，包括高灵敏度和特异性、实时监测能力以及实验设计的多功能性。在本章中，我们概述了基于荧光蛋白的FRET分析（包括单分子FRET，即smFRET）在病毒学研究中的应用程序，并提供故障排除说明。
