The uses of green fluorescent protein in mammalian cells.
The uses of green fluorescent protein in mammalian cells.
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绿色荧光蛋白在哺乳动物细胞中的用途。
DOI：
10.1002/0471739499.ch14
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发表时间：
2006
期刊：
影响因子：
--
通讯作者：
Lippincott-Schwartz,Jennifer
中科院分区：
文献类型：
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作者：
Ward,TheresaH;Lippincott-Schwartz,Jennifer
绿色荧光蛋白 (GFP) 嵌合体在细胞行为和动力学研究中的应用现已普遍存在于哺乳动物细胞生物学的所有领域。这种广泛的应用是由于 (a) 新 GFP 变体的开发和优化的细胞表达策略，可产生明亮、稳定的荧光信号，以及 (b) 荧光成像方法和显微镜系统的进步，使分析活细胞中的蛋白质地理、运动和化学变得简单。在这里，我们讨论了几种基于 GFP 的技术，包括延时成像、光漂白、光活化和荧光共振能量转移 (FRET)，这些技术允许在活哺乳动物细胞中分析蛋白质动力学、功能和表达。我们进一步描述了这些技术如何识别出哺乳动物细胞稳态所必需的新途径和机制，而传统的生化方法无法解决这些问题。
The use of green fluorescent protein (GFP) chimeras in the study of cell behavior and dynamics is now ubiquitous in all fields of mammalian cell biology. This extensive use is due to (a) the development of new GFP variants and optimized cell expression strategies that produce bright, stable fluorescent signals and (b) advances in fluorescent imaging methods and microscopy systems that make it simple to analyze protein geography, movement, and chemistry in living cells. Here, we discuss several GFP-based techniques including time-lapse imaging, photobleaching, photoactivation, and fluorescence resonance energy transfer (FRET) that have allowed protein dynamics, function, and expression to be analyzed in living mammalian cells. We further describe how these techniques have led to the identification of new pathways and mechanisms essential for mammalian cell homeostasis, which traditional biochemical approaches have been unable to address.