UNDERSTANDING, IMPROVING AND USING GREEN FLUORESCENT PROTEINS
UNDERSTANDING, IMPROVING AND USING GREEN FLUORESCENT PROTEINS
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DOI：
10.1016/s0968-0004(00)89099-4
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发表时间：
1995-11-01
影响因子：
13.8
通讯作者：
TSIEN, RY
中科院分区：
文献类型：
--
作者：
CUBITT, AB;HEIM, R;TSIEN, RY
绿色荧光蛋白（GFP）作为通过纯分子生物学手段产生强可见荧光的第一种通用方法目前吸引了巨大的兴趣。到目前为止，它们已被用作基因表达的报告者，细胞谱系的示踪剂，以及作为融合标签来监测活细胞内的蛋白质定位，然而，最初从水母Aequorea维多利亚克隆的GFP具有几个非最佳特性，包括低亮度，蛋白质合成和荧光发育之间的显着延迟，以及复杂的光异构化。幸运的是，这种蛋白质可以通过诱变来重新设计，以改善这些缺陷，并改变激发和发射波长，创造不同的颜色和新的应用。
Green fluorescent proteins (GFPs) are presently attracting tremendous interest as the first general method to create strong visible fluorescence by purely molecular biological means. So far, they have been used as reporters of gene expression, tracers of cell lineage, and as fusion tags to monitor protein localization within living cells, However, the GFP originally cloned from the jellyfish Aequorea victoria has several nonoptimal properties including low brightness, a significant delay between protein synthesis and fluorescence development, and complex photoisomerization. Fortunately, the protein can be re-engineered by mutagenesis to ameliorate these deficiencies and shift the excitation and emission wavelengths, creating different colors and new applications.