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全细胞细菌传感器在生物技术和环境科学中的应用。

Applications of whole-cell bacterial sensors in biotechnology and environmental science.

作者信息

Yagi Kiyohito

机构信息

Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamada-oka, Suita, Osaka, 565-0871, Japan.

出版信息

Appl Microbiol Biotechnol. 2007 Jan;73(6):1251-8. doi: 10.1007/s00253-006-0718-6. Epub 2006 Nov 17.

Abstract

Biosensors have major advantages over chemical or physical analyses with regard to specificity, sensitivity, and portability. Recently, many types of whole-cell bacterial biosensors have been developed using recombinant DNA technology. The bacteria are genetically engineered to respond to the presence of chemicals or physiological stresses by synthesizing a reporter protein, such as luciferase, beta-galactosidase, or green fluorescent protein. In addition to an overview of conventional biosensors, this minireview discusses a novel type of biosensor using a photosynthetic bacterium as the sensor strain and the crtA gene, which is responsible for carotenoid synthesis, as the reporter. Since bacteria possess a wide variety of stress-response mechanisms, including antioxidation, heat-shock responses, nutrient-starvation, and membrane-damage responses, DNA response elements for several stress-response proteins can be fused with various reporter genes to construct a versatile set of bacterial biosensors for a variety of analytes. Portable biosensors for on-site monitoring have been developed using a freeze-dried biosensing strain, and cell array biosensors have been designed for high-throughput analysis. Moreover, in the future, the use of single-cell biosensors will permit detailed analyses of samples. Signals from such sensors could be detected with digital imaging, epifluorescence microscopy, and/or flow cytometry.

摘要

生物传感器在特异性、灵敏度和便携性方面比化学或物理分析具有显著优势。最近,利用重组DNA技术开发了多种类型的全细胞细菌生物传感器。通过基因工程改造细菌,使其在检测到化学物质或生理应激时合成报告蛋白,如荧光素酶、β-半乳糖苷酶或绿色荧光蛋白。除了对传统生物传感器进行概述外,本综述还讨论了一种新型生物传感器,该传感器使用光合细菌作为传感菌株,并使用负责类胡萝卜素合成的crtA基因作为报告基因。由于细菌具有多种应激反应机制,包括抗氧化、热休克反应、营养饥饿和膜损伤反应,几种应激反应蛋白的DNA反应元件可以与各种报告基因融合,构建一套通用的用于多种分析物的细菌生物传感器。利用冻干的生物传感菌株开发了用于现场监测的便携式生物传感器,并设计了用于高通量分析的细胞阵列生物传感器。此外,未来单细胞生物传感器的使用将允许对样品进行详细分析。可以通过数字成像、落射荧光显微镜和/或流式细胞术检测来自此类传感器的信号。

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