A LuxP-based fluorescent sensor for bacterial autoinducer II.
A LuxP-based fluorescent sensor for bacterial autoinducer II.
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DOI：
10.1021/cb7002048
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发表时间：
2008-01
影响因子：
4
通讯作者：
Jinge G. Zhu;D. Pei
中科院分区：
文献类型：
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作者：
Jinge G. Zhu;D. Pei
自诱导子 2 (AI-2) 使不同细菌物种能够进行种间通讯，但一直难以定量检测。目前，最常用的 AI-2 检测方法是采用工程化的哈维氏弧菌报告菌株，该菌株可响应 AI-2 产生生物发光。然而，生物测定不是定量的并且对测定条件敏感。在这项工作中，我们通过使用环境敏感的荧光染料修饰AI-2受体蛋白LuxP和LsrB，开发了两种AI-2蛋白传感器。蛋白质传感器与 AI-2 特异性结合，并使其荧光产量产生剂量依赖性变化。新的检测方法已用于实时监测AI-2的酶促合成，并测定多种细菌培养液中细胞外和细胞内AI-2的浓度。
Autoinducer 2 (AI-2), which enables different bacterial species to engage in interspecies communication, has been difficult to detect quantitatively. Currently, the most commonly used method for AI-2 detection employs an engineered Vibrio harveyi reporter strain, which produces bioluminescence in response to AI-2. However, the bioassay is not quantitative and is sensitive to assay conditions. In this work, we have developed two protein sensors for AI-2 by modifying AI-2 receptor proteins LuxP and LsrB with environmentally sensitive fluorescent dyes. The protein sensors bind specifically to AI-2 and produce dose-dependent changes in their fluorescence yield. The new assay method has been applied to monitor the enzymatic synthesis of AI-2 in real time and determine the extracellular and intracellular AI-2 concentrations in several bacterial culture fluids.