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The green fluorescent protein as a visible marker for quantifying thehydrogen-producing bacterium of Enterobacter aerogenes in an anaerobiccomplex system

机译：绿色荧光蛋白作为可见标记，用于定量厌氧复合系统中产气肠杆菌产氢细菌

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摘要

The potential of the gfp (green fluorescent protein) gene from the jellyfish Aequoria victoriaas a visible marker for tracking hydrogen-producing bacteria Enterobacter aerogenes in ananaerobic complex culture system was examined.A gfp-and kanamycin-resistance gene-containing plasmid pUCGK was constructed andtransformed into E. aerogenes. The fluorescence resulting from the green fluorescent protein(GFP) expression was detected in this fluorescent bacterium under different conditions.Since the use of GFP as a molecular reporter is restricted by its requirement for oxygen in thedevelopment of the fluorophore, fluorescence detection for the fluorescent strain grownaerobically and anaerobically was performed respectively. The oxygen concentration formaturation of the GFP fluorophore in E. aerogenes(pUCGK) and the fluorescence detectioncondition for the fluorescent strain were determined in the hydrogen production system in thepresent study.By using the GFP technique, a method for rapid and non-disruptive cell quantification of E.Aerogenes by fluorescence density was developed to analyze the hydrogen production process.Cell dry weight of the strain could be easily achieved through the fluorescence intensitydetection. These findings allow the gfp-harboring strain of E. aerogenes to be quantified in ananaerobic complex culture system for enhanced hydrogen production. As far as we know, weare the first to apply gfp to E. aerogenes for analyzing the hydrogen production process.
机译：研究了水母维多利亚水母的gfp（绿色荧光蛋白）基因作为追踪厌氧复合培养系统中产氢细菌产气肠杆菌的可见标记的潜力。构建并转化了含gfp和卡那霉素抗性基因的质粒pUCGK进入产气大肠杆菌。在不同条件下，在该荧光细菌中检测到由绿色荧光蛋白（GFP）表达产生的荧光。由于使用GFP作为分子报告分子受到荧光团发展中对氧气的需求的限制，因此对该荧光菌株进行荧光检测分别进行厌氧和厌氧生长。本研究在制氢系统中确定了产气大肠杆菌（pUCGK）中GFP荧光团的氧浓度形成和荧光菌株的荧光检测条件。通过GFP技术，一种快速无中断的细胞定量方法利用荧光密度法对大肠杆菌进行了分析，以分析制氢过程。通过荧光强度检测可以很容易地获得菌株的细胞干重。这些发现允许在厌氧复合培养系统中定量产气链球菌的gfp菌株以提高产氢量。据我们所知，是第一个将gfp应用于产气大肠杆菌分析制氢过程的公司。

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