Flavin Mononucleotide-Based Fluorescent Reporter Proteins Outperform Green Fluorescent Protein-Like Proteins as Quantitative In Vivo Real-Time Reporters
Flavin Mononucleotide-Based Fluorescent Reporter Proteins Outperform Green Fluorescent Protein-Like Proteins as Quantitative In Vivo Real-Time Reporters
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DOI：
10.1128/aem.00701-10
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发表时间：
2010-09-01
影响因子：
4.4
通讯作者：
Jaeger, Karl-Erich
中科院分区：
文献类型：
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作者：
Drepper, Thomas;Huber, Robert;Jaeger, Karl-Erich
绿色荧光蛋白（GFP）家族的荧光蛋白通常用作报告蛋白，用于定量分析活微生物中的复杂生物过程。在这里，我们证明了GFP样蛋白的荧光信号强度受氧限制，因此并不反映大肠杆菌分批培养的报告蛋白的量。相反，黄素单核苷酸（FMN）结合荧光蛋白（FbFP）是适合在这些条件下的定量实时体内测定。在过去的十年里，
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. In the last decade,