Fluorescent proteins for FRET microscopy: monitoring protein interactions in living cells.

Fluorescent proteins for FRET microscopy: monitoring protein interactions in living cells.
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用于FRET显微镜的荧光蛋白:监测活细胞中的蛋白质相互作用。

DOI:
10.1002/bies.201100098
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发表时间:
2012-05
期刊:
影响因子:
4
通讯作者:
Davidson, Michael W.
Davidson, Michael W.
中科院分区:
生物学3区
文献类型:
--
作者:
Day, Richard N.;Davidson, Michael W.

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来自不同生物体的新型荧光蛋白(FP)的发现和工程正在产生具有特殊特性的活细胞成像荧光团。特别是,用于荧光(或Förster)共振能量转移(FRET)显微镜的FP的开发为监测活细胞内动态蛋白质相互作用提供了重要工具。在FRET显微镜的兴趣增加,推动了许多不同的方法来测量FRET的发展。然而,FRET测量的解释是复杂的几个因素,包括高荧光背景,潜在的光转换文物,和相对较低的动态范围提供了这种技术。在这里,我们描述的优点和缺点,四种方法中常用的FRET显微镜。然后,我们讨论了选择不同的FRET方法的FP,确定最有用的FP候选FRET显微镜。最近成功地扩大FP调色板提供了机会,探索新的FRET对。
The discovery and engineering of novel fluorescent proteins (FPs) from diverse organisms is yielding fluorophores with exceptional characteristics for live-cell imaging. In particular, the development of FPs for fluorescence (or Förster) resonance energy transfer (FRET) microscopy is providing important tools for monitoring dynamic protein interactions inside living cells. The increased interest in FRET microscopy has driven the development of many different methods to measure FRET. However, the interpretation of FRET measurements is complicated by several factors including the high fluorescence background, the potential for photoconversion artifacts, and the relatively low dynamic range afforded by this technique. Here, we describe the advantages and disadvantages of four methods commonly used in FRET microscopy. We then discuss the selection of FPs for the different FRET methods, identifying the most useful FP candidates for FRET microscopy. The recent success in expanding the FP color palette offers the opportunity to explore new FRET pairs.
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