GFP imaging:: methodology and application to investigate cellular compartmentation in plants
GFP imaging:: methodology and application to investigate cellular compartmentation in plants
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DOI：
10.1093/jexbot/52.356.529
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发表时间：
2001-04-01
影响因子：
6.9
通讯作者：
Köhler, RH
中科院分区：
文献类型：
--
作者：
Hanson, MR;Köhler, RH
水母绿色荧光蛋白(GFP)基因的克隆及其在转化植物细胞中亚细胞位置表达的改变，导致了对细胞内组织和动力学的新观点。GFP与已知或未知功能的完整蛋白质的融合显示了蛋白质的位置以及蛋白质是否从一个隔室移动到另一个隔室。绿色荧光蛋白和具有不同光谱特性的变体被故意定位于分开的隔室，以确定它们的大小、形状、流动性和在开发或环境响应期间的动态变化。绿色荧光蛋白变异体之间的荧光共振能量转移(FRET)可以识别蛋白质/蛋白质之间的相互作用。GFP已被用作传感器来检测钙、pH、电压、金属和酶活性的变化或差异。GFP的光漂白和光激活以及荧光相关光谱可以测量GFP在隔室内或之间的扩散和移动速度。本文综述了这些方法的过去应用，以及GFP成像在理解植物细胞功能组织方面的有希望的进展。
The cloning of the jellyfish gfp (green fluorescent protein) gene and its alteration for expression in subcellular locations in transformed plant cells have resulted in new views of intracellular organization and dynamics. Fusions of GFP with entire proteins of known or unknown function have shown where the proteins are located and whether the proteins move from one compartment to another. GFP and variants with different spectral properties have been deliberately targeted to separate compartments to determine their size, shape, mobility, and dynamic changes during development or environmental response. Fluorescence Resonance Energy Transfer (FRET) between GFP variants can discern protein/protein interactions. GFP has been used as a sensor to detect changes or differences in calcium, pH, voltage, metal, and enzyme activity. Photobleaching and photoactivation of GFP as well as fluorescence correlation spectroscopy can measure rates of diffusion and movement of GFP within or between compartments. This review covers past applications of these methods as well as promising developments in GFP imaging for understanding the functional organization of plant cells.