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首页> 外文期刊>Journal of structural and functional genomics >Recent Advances in GFP Folding Reporter and Split-GFP Solubility Reporter Technologies. Application to Improving the Folding and Solubility of Recalcitrant Proteins from Mycobacterium tuberculosis
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Recent Advances in GFP Folding Reporter and Split-GFP Solubility Reporter Technologies. Application to Improving the Folding and Solubility of Recalcitrant Proteins from Mycobacterium tuberculosis

机译：GFP Folding Reporter和Split-GFP Solubility Reporter技术的最新进展。在改善结核分枝杆菌顽calc蛋白的折叠性和溶解性中的应用

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We have improved our green fluorescent protein (GFP) folding reporter technology [Waldo et al., (1999) Nat. Biotechnol. 17, 691–695] to evolve recalcitrant proteins from Mycobacterium tuberculosis. The target protein is inserted into the scaffolding of the GFP, eliminating false-positive artifacts caused by expression of truncated protein variants from internal cryptic ribosome binding sites in the target RNA. In parallel, we have developed a new quantitative fluorescent protein tagging and detection system based on micro-domains of GFP. This split-GFP system, which works both in vivo and in vitro, is amenable to high-throughput assays of protein expression and solubility [Cabantous et al., (2005) Nat. Biotechnol. 23, 102–107]. Together, the GFP folding reporter and split-GFP technologies offer a comprehensive system for manipulating and improving protein folding and solubility.
机译：我们已经改进了绿色荧光蛋白（GFP）折叠报道基因技术[Waldo等，（1999）Nat。生物技术。 17，17，691–695]从结核分枝杆菌进化出顽固蛋白。将靶蛋白插入GFP的支架中，消除了由靶RNA中内部隐性核糖体结合位点表达截短的蛋白变体引起的假阳性假象。同时，我们开发了一种新的基于GFP微域的定量荧光蛋白标记和检测系统。该分裂的GFP系统在体内和体外均起作用，适用于蛋白质表达和溶解度的高通量测定[Cabantous等人，（2005）Nat.Natl.Acad.Sci.USA，87：1593-1877]。生物技术。 23，102–107]。 GFP折叠报告基因和split-GFP技术一起提供了用于操纵和改善蛋白质折叠和溶解度的综合系统。

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