Terskikh A, Fradkov A, Ermakova G, Zaraisky A, Tan P, Kajava A V, Zhao X, Lukyanov S, Matz M, Kim S, Weissman I, Siebert P
School of Medicine, Stanford University, Stanford, CA 94305, USA.
Science. 2000 Nov 24;290(5496):1585-8. doi: 10.1126/science.290.5496.1585.
We generated a mutant of the red fluorescent protein drFP583. The mutant (E5) changes its fluorescence from green to red over time. The rate of color conversion is independent of protein concentration and therefore can be used to trace time-dependent expression. We used in vivo labeling with E5 to measure expression from the heat shock-dependent promoter in Caenorhabditis elegans and from the Otx-2 promoter in developing Xenopus embryos. Thus, E5 is a "fluorescent timer" that can be used to monitor both activation and down-regulation of target promoters on the whole-organism scale.
我们生成了红色荧光蛋白drFP583的一个突变体。该突变体（E5）的荧光会随时间从绿色转变为红色。颜色转换的速率与蛋白质浓度无关，因此可用于追踪时间依赖性表达。我们使用E5进行体内标记，以测量秀丽隐杆线虫中热休克依赖性启动子以及非洲爪蟾发育胚胎中Otx - 2启动子的表达。因此，E5是一种“荧光定时器”，可用于在全生物体尺度上监测靶启动子的激活和下调。
