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A novel method to analyze nucleocytoplasmic transport in vivo by using short peptide tags

机译:一种使用短肽标签分析体内核质转运的新方法

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Regulated nucleocytoplasmic transport is of vital importance for maintaining the physiology of the cell, and disturbed nucleocytoplasmic shuttling of certain proteins has been found in a variety of diseases including cancer. The most frequently used procedure to analyze those processes is to fuse the protein of interest to a fluorescent protein such as GFP (green fluorescent protein) - a technique that is prone to impair normal protein function and subcellular localization. We report a novel approach to monitor nucleocytoplasmic transport processes in vivo by combining short TetR inducing peptide tags (TIP) with a TetR-controlled reporter gene in a human cell line. The technology is exemplified by demonstrating nucleocytoplasmic shuttling of the glucocorticoid receptor and activity of two further TIP fusions to cancer-related proteins. The technology presented provides the basis for efficient screening systems to isolate compounds altering the nucleocytoplasmic distribution of a protein of interest.
机译:调节性的核质转运对于维持细胞的生理至关重要,并且在包括癌症在内的多种疾病中发现了某些蛋白质的核质穿梭紊乱。分析这些过程的最常用方法是将目标蛋白与荧光蛋白(例如GFP(绿色荧光蛋白))融合-这种技术容易损害正常的蛋白功能和亚细胞定位。我们报告一种新型的方法,通过结合人类细胞系中的TetR控制的报道基因短TetR诱导肽标签(TIP)来监测体内的核质运输过程。该技术通过展示糖皮质激素受体的核质穿梭作用以及两种另外的TIP融合蛋白与癌症相关蛋白的活性来进行例证。提出的技术为有效的筛选系统提供了基础,以分离出改变目标蛋白质胞质分布的化合物。

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