Abstract：
Fluorescent proteins, such as green fluorescent proteins, are invaluable tools for detecting and quantifying gene expression in high-throughput reporter gene assays. However, they introduce significant inaccuracies in studies involving microaerobiosis or anaerobiosis, as oxygen is required for the maturation of these proteins\' chromophores. In this study, the authors highlight the errors incurred by using fluorescent proteins under limited oxygenation by comparing standard fluorescence-based reporter gene assays to quantitative real-time PCR data in the study of a complex oxygen-regulated gene network. Furthermore, a solution to perform quantification of anaerobic and microaerobic gene expression with fluorescent reporter proteins using a microplate reader with an oxygen control system and applying pulses of full oxygenation before fluorescence measurements is provided.
摘要：
荧光蛋白,如绿色荧光蛋白，是在高通量报告基因测定中检测和定量基因表达的宝贵工具。然而,它们在涉及微需氧菌或厌氧菌的研究中引入了重大的不准确性，因为氧气是这些蛋白质发色团成熟所必需的。在这项研究中,作者通过在复杂的氧调节基因网络的研究中比较标准的基于荧光的报告基因测定与定量实时PCR数据,强调了在有限氧合下使用荧光蛋白所导致的错误.此外，提供了一种使用具有氧气控制系统的微孔板读数器并在荧光测量之前施加完全氧合脉冲用荧光报告蛋白进行厌氧和微需氧基因表达定量的解决方案。
