Photoactivation turns green fluorescent protein red
Photoactivation turns green fluorescent protein red
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DOI：
10.1016/s0960-9822(06)00342-3
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发表时间：
1997-10-01
期刊：
影响因子：
9.2
通讯作者：
Leibler, S
中科院分区：
文献类型：
--
作者：
Elowitz, MB;Surette, MG;Leibler, S
在其基因首次被克隆后的几年中，维多利亚水母绿色荧光蛋白（GFP）已成为细胞生物学中的有力工具，作为活细胞中基因表达、蛋白定位和蛋白动力学的标记物[1-3]，已鉴定出具有改善的荧光强度和改变的光谱特征的GFP变体，但是对于多种标记实验、蛋白质相互作用研究和在某些生物体中提高可见度，仍然需要另外的GFP变体[4]。特别地，长波长（红色）荧光仍然是难以捉摸的。这里我们描述了一种红色发射，绿色荧光蛋白的绿色吸收荧光状态是由蓝光光活化产生的，在低氧浓度的条件下，绿色荧光蛋白可以很容易地用激光或荧光显微镜灯转换到其红色发射状态，这种以前未被注意到的能力使得能够在体内对蛋白质进行区域性、非侵入性标记。特别是，我们在这里报告使用绿色荧光蛋白的光活化，使第一个直接测量蛋白质扩散在细胞质中的活细菌。(C)Current Biology Ltd ISSN 0960-9822。
In the few years since its gene was first cloned, the Aequorea victoria green fluorescent protein (GFP) has become a powerful tool in cell biology, functioning as a marker for gene expression, protein localization and protein dynamics in living cells [1-3], GFP variants with improved fluorescence intensity and altered spectral characteristics have been identified, but additional GFP variants are still desirable for multiple labeling experiments, protein interaction studies and improved visibility in some organisms [4], In particular, long-wavelength (red) fluorescence has remained elusive, Here we describe a red emitting, green-absorbing fluorescent state of GFP that is generated by photoactivation with blue light, GFP can be switched to its red-emitting state easily with a laser or fluorescence microscope lamp under conditions of low oxygen concentration, This previously unnoticed ability enables regional, non-invasive marking of proteins in vivo. In particular, we report here the use of GFP photoactivation to make the first direct measurements of protein diffusion in the cytoplasm of living bacteria. (C) Current Biology Ltd ISSN 0960-9822.