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A noncytotoxic DsRed variant for whole-cell labeling

机译:一种无细胞毒性的DsRed变体,用于全细胞标记

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摘要

Fluorescent proteins (FPs) are extremely useful tools for whole-cell, tissue, and animal labeling. For these purposes, FPs may be monomeric or oligomeric, but should meet the criteria of being tolerated at high expression levels in cells and having desirable photophysical properties.rnOur goal was to create a variant of DsRed-Express that maintains the brightness, fast-maturation, and photostability of this protein, while exhibiting decreased cytotoxicity. For this purpose, we mutated the surface of DsRed-Express to decrease aggregation and created DsRed-Express2. DsRed-Express2 retains the favorable photophysical properties of DsRed-Express while showing dramatically reduced cytotoxicity and higher expression in bacterial and mammalian systems. Further, it was shown that DsRed-Express2 outperforms other red FPs as a label for bacterial and mammalian cells.
机译:荧光蛋白(FPs)是用于全细胞,组织和动物标记的极其有用的工具。为此,FPs可以是单体或寡聚的,但应满足在细胞中高表达水平下具有耐受性并具有理想的光物理特性的标准。​​我们的目标是创建一种DsRed-Express变体,以保持亮度,快速成熟。和这种蛋白质的光稳定性,同时表现出降低的细胞毒性。为此,我们对DsRed-Express的表面进行了突变以减少聚集,并创建了DsRed-Express2。 DsRed-Express2保留了DsRed-Express的良好光物理特性,同时在细菌和哺乳动物系统中显示出大大降低的细胞毒性和更高的表达。此外,已表明,DsRed-Express2优于其他红色FP作为细菌和哺乳动物细胞的标记。

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