含Dsred类似生色团的红色荧光蛋白的发展及应用

自从绿色荧光蛋白(GFP)被发现以来，荧光蛋白在生物医学领域已经成为一种重要的荧光成像工具．随着红色荧光蛋白DsRed的出现，各种优化的DsRed突变体和远红荧光蛋白也不断涌现．其中荧光蛋白生色团的形成机制对改建更优的荧光蛋白变种影响很大，对于红色荧光蛋白而言，大多数的红色荧光蛋白的生色团类型为DsRed类似生色团，在此基础上又出现了Far-red DsRed类似生色团．目前，含DsRed类似生色团的荧光蛋白主要有单体红色荧光蛋白、光转换荧光蛋白、斯托克斯红移蛋白、荧光计时器等．这些优化的荧光蛋白作为分子探针可以实现对活细胞、细胞器或胞内分子的时空标记和追踪，已经在生物工程学、细胞生物学、基础医学领域得到广泛应用．本文综述了含DsRed类似生色团的荧光蛋白的研究进展及其应用，以及由此发展起来的远红荧光蛋白在活体显微成像技术中的应用，并展望了荧光探针技术研究的新方向．

The Development and Appliation of Red Fluorescent Proteins with DsRed-like Chromophore

Fluorescent proteins (FPs), represented by green fluorescent protein (GFP), are invaluable tools for biomedical research. The discovery of DsRed led to the successful development of other new red fluorescent proteins and far-red fluorescent proteins and the applications in non-bioluminescent organisms. The chromophore maturation process of fluorescent proteins is important for their optimization,especially for the widely red fluorescent proteins (RFPs) known are the DsRed-like chromophore. Far-red DsRed-like chromophore, such as monomeric RFPs, photoswitchable FPs, large stoke shift FPs, and fluorescent timers, has been developed based on the DsRed mechanism. These mutants from DsRed-like chromophore are being used as the molecular probes for noninvasive labeling and tracking of proteins, organelles, and cells in real time, and showed great potential applications in biotechnology, cell biology and preclinical medicine. This review summarized the recent progress and application of RFPs of DsRed-like chromophore, the far-red fluorescent proteins designed for deep-tissue and whole-animal imaging as well as probes in flow cytometry and fluorescence microscopy.