A cell-free fluorescence biosensor based on allosteric transcription factor NalC for detection of pentachlorophenol.

IF 2 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Biotechnology Letters Pub Date : 2024-10-01 Epub Date: 2024-07-17 DOI:10.1007/s10529-024-03511-1

Shuting Chen, Chen Zhao, Xiaodan Kang, Xi Zhang, Bin Xue, Chenyu Li, Shang Wang, Xiaobo Yang, Chao Li, Zhigang Qiu, Jingfeng Wang, Zhiqiang Shen

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Abstract

Pentachlorophenol (PCP) was once used as a pesticide, germicide, and preservative due to its stable properties and resistance to degradation. This study aimed to design a biosensor for the quantitative and prompt detection of capable of PCP. A cell-free fluorescence biosensor was developed while employing NalC, an allosteric Transcription Factor responsive to PCP and In Vitro Transcription. By adding a DNA template and PCP and employing Electrophoretic Mobility Shift Assay while monitoring the dynamic fluorescence changes in RNA, this study offers evidence of NalC's potential applicability in sensor systems developed for the specific detection of PCP. The biosensor showed the capability for the quantitative detection of PCP, with a Limit of Detection (LOD) of 0.21 μM. Following the addition of Nucleic Acid Sequence-Based Amplification, the fluorescence intensity of RNA revealed an excellent linear relationship with the concentration of PCP, showing a correlation coefficient (R²) of 0.9595. The final LOD was determined to be 0.002 μM. This study has successfully translated the determination of PCP into a fluorescent RNA output, thereby presenting a novel approach for detecting PCP within environmental settings.

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基于异位转录因子 NalC 的无细胞荧光生物传感器，用于检测五氯苯酚。

五氯苯酚（PCP）因其性质稳定、耐降解，曾被用作杀虫剂、杀菌剂和防腐剂。本研究旨在设计一种生物传感器，用于定量和快速检测五氯苯酚的能力。研究人员利用 NalC（一种对五氯苯酚有反应的异位转录因子）和体外转录技术，开发了一种无细胞荧光生物传感器。通过添加 DNA 模板和五氯苯酚，并采用电泳迁移分析法监测 RNA 的动态荧光变化，该研究证明了 NalC 在开发用于特异性检测五氯苯酚的传感器系统中的潜在适用性。该生物传感器具有定量检测五氯苯酚的能力，检测限（LOD）为 0.21 μM。加入核酸序列扩增后，RNA 的荧光强度与五氯苯酚的浓度呈良好的线性关系，相关系数 (R2) 为 0.9595。最终的检测限被确定为 0.002 μM。这项研究成功地将五氯苯酚的测定转化为荧光 RNA 输出，从而为在环境中检测五氯苯酚提供了一种新方法。

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来源期刊

Biotechnology Letters 工程技术-生物工程与应用微生物

CiteScore

5.90

自引率

3.70%

发文量

108

审稿时长

1.2 months

期刊介绍： Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.