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黄素单核苷酸基荧光报告蛋白在作为定量活体实时报告基因方面优于绿色荧光蛋白样蛋白。

Flavin mononucleotide-based fluorescent reporter proteins outperform green fluorescent protein-like proteins as quantitative in vivo real-time reporters.

机构信息

Institute of Molecular Enzyme Technology, Heinrich Heine University Duesseldorf, Research Center Juelich, Stetternicher Forst, D-52426 Juelich, Germany.

出版信息

Appl Environ Microbiol. 2010 Sep;76(17):5990-4. doi: 10.1128/AEM.00701-10. Epub 2010 Jul 2.

Abstract

Fluorescent proteins of the green fluorescent protein (GFP) family are commonly used as reporter proteins for quantitative analysis of complex biological processes in living microorganisms. Here we demonstrate that the fluorescence signal intensity of GFP-like proteins is affected under oxygen limitation and therefore does not reflect the amount of reporter protein in Escherichia coli batch cultures. Instead, flavin mononucleotide (FMN)-binding fluorescent proteins (FbFPs) are suitable for quantitative real-time in vivo assays under these conditions.

摘要

绿色荧光蛋白(GFP)家族的荧光蛋白通常被用作报告蛋白,用于定量分析活微生物中复杂的生物过程。在这里,我们证明 GFP 样蛋白的荧光信号强度会受到缺氧的影响,因此不能反映大肠杆菌分批培养物中报告蛋白的数量。相反,黄素单核苷酸(FMN)结合荧光蛋白(FbFPs)在这些条件下适合用于定量实时体内测定。

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本文引用的文献

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Reporter proteins for in vivo fluorescence without oxygen.用于无氧条件下体内荧光的报告蛋白。
Nat Biotechnol. 2007 Apr;25(4):443-5. doi: 10.1038/nbt1293. Epub 2007 Mar 11.
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