Swammerdam Institute for Life Sciences, Section of Molecular Cytology, Centre for Advanced Microscopy, University of Amsterdam, Amsterdam, The Netherlands.
Nat Methods. 2010 Feb;7(2):137-9. doi: 10.1038/nmeth.1415. Epub 2010 Jan 17.
Optimization of autofluorescent proteins by intensity-based screening of bacteria does not necessarily identify the brightest variant for eukaryotes. We report a strategy to screen excited state lifetimes, which identified cyan fluorescent proteins with long fluorescence lifetimes (>3.7 ns) and high quantum yields (>0.8). One variant, mTurquoise, was 1.5-fold brighter than mCerulean in mammalian cells and decayed mono-exponentially, making it an excellent fluorescence resonance energy transfer (FRET) donor.
通过基于强度的细菌筛选对自发荧光蛋白进行优化，不一定能鉴定出真核生物中最亮的变体。我们报告了一种筛选激发态寿命的策略，该策略鉴定出了具有长荧光寿命（>3.7 ns）和高量子产率（>0.8）的青色荧光蛋白。一种变体 mTurquoise 在哺乳动物细胞中的亮度比 mCerulean 高 1.5 倍，且呈单指数衰减，使其成为一种出色的荧光共振能量转移（FRET）供体。
