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利用合成 CadC-T7 遗传线路开发高特异性和高灵敏度的镉和铅微生物生物传感器。

Development of a highly specific and sensitive cadmium and lead microbial biosensor using synthetic CadC-T7 genetic circuitry.

机构信息

Infection and Immunity Research Center, Korea Research Institute Bioscience and Biotechnology (KRIBB), Daejeon, Republic of Korea; Biosystems & Bioengineering Program, University of Science and Technology (UST), Daejeon, Republic of Korea.

Department of Biomedical Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan, Republic of Korea.

出版信息

Biosens Bioelectron. 2016 May 15;79:701-8. doi: 10.1016/j.bios.2015.12.101. Epub 2015 Dec 29.

Abstract

Multiple copies of a cadC homolog encoding a heavy metal-responsive transcription factor were found in the genome of a bacterium isolated from ocean sediment, and the heavy metal responses of the encoded proteins were characterized using a fluorescence reporter assay. Each CadC regulator exhibited distinct specificity in response to heavy metal ions, indicating their potential use as modular heavy metal biosensors. Next, we constructed CadC-controlled T7 RNA transcription systems for intracellular signal amplification, i.e., higher sensitivity. Flow cytometry revealed that cadmium and lead ions could be recognized specifically by CadC-T7 biosensors, which could be combined with a microfluidic platform to generate heavy metal biosensor devices with increased sensitivity. Our results demonstrate the successful development of synthetic CadC-T7 genetic circuitry for use in improved heavy metal biosensor microfluidic devices.

摘要

从海洋沉积物中分离出的一种细菌的基因组中发现了多个 cadC 同源物拷贝,这些拷贝编码重金属应答转录因子。使用荧光报告测定法对编码蛋白的重金属应答特性进行了表征。每个 CadC 调节剂对重金属离子的应答都具有独特的特异性,表明它们有可能作为模块化重金属生物传感器使用。接下来,我们构建了 CadC 控制的 T7 RNA 转录系统,用于细胞内信号放大,即提高灵敏度。流式细胞术显示 CadC-T7 生物传感器可以特异性识别镉和铅离子,这些生物传感器可以与微流控平台结合,生成灵敏度更高的重金属生物传感器装置。我们的研究结果证明了合成 CadC-T7 遗传电路在改进的重金属生物传感器微流控装置中的成功应用。

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