Oxidative stress detection with Escherichia coli harboring a katG'::lux fusion
Oxidative stress detection with Escherichia coli harboring a katG'::lux fusion
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DOI：
10.1128/aem.62.7.2252-2256.1996
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发表时间：
1996-07-01
影响因子：
4.4
通讯作者：
LaRossa, RA
中科院分区：
文献类型：
--
作者：
Belkin, S;Smulski, DR;LaRossa, RA
含有大肠杆菌katG启动子与截短的费氏弧菌lux操纵子的转录融合体的质粒（luxCDABE）构建了一个带有该质粒的大肠杆菌菌株，（菌株DPD 2511）表现出低的发光基础水平，在过氧化氢、有机过氧化物、氧化还原循环剂的存在下，发光基础水平增加高达1,000倍。（甲基紫精和甲萘醌），过氧化氢产生酶系统（黄嘌呤和黄嘌呤氧化酶）和香烟烟雾。oxyR缺失消除了过氧化氢依赖性诱导，证实oxyR控制katG '：Lux发光，乙醇也通过一种无法解释的机制诱导发光，当细胞暴露于乙醇和过氧化氢时，观察到显著的协同反应;在两种诱导剂存在下测得的发光水平远高于用乙醇观察到的发光水平和用氢观察到的发光水平之和建议这种结构或类似结构可用作测定化学品的氧化剂和抗氧化剂性质的工具，用作环境监测的生物传感器，以及用作研究细胞对氧化危害的反应的工具。
A plasmid containing a transcriptional fusion of the Escherichia coli katG promoter to a truncated Vibrio fischeri lux operon (luxCDABE) was constructed, An E, coli strain bearing this plasmid (strain DPD2511) exhibited low basal levels of luminescence, which increased up to 1,000-fold in the presence of hydrogen peroxide, organic peroxides, redox-cycling agents (methyl viologen and menadione), a hydrogen peroxide producing enzyme system (xanthine and xanthine oxidase), and cigarette smoke, An oxyR deletion abolished hydrogen peroxide-dependent induction, confirming that oxyR controlled katG'::Lux luminescence, Light emission was also induced by ethanol by an unexplained mechanism, A marked synergistic response was observed when cells were exposed to both ethanol and hydrogen peroxide; the level of luminescence measured in the presence of both inducers was much higher than the sum of the level of luminescence observed with ethanol and the level of luminescence observed with hydrogen peroxide, It is suggested that this construction or similar constructions may be used as a tool for assaying oxidant and antioxidant properties of chemicals, as a biosensor for environmental monitoring, and as a tool for studying cellular responses to oxidative hazards.