Development and Application of a Synthetically-Derived Lead Biosensor Construct for Use in Gram-Negative Bacteria.
Development and Application of a Synthetically-Derived Lead Biosensor Construct for Use in Gram-Negative Bacteria.
复制标题
DOI：
10.3390/s16122174
复制
发表时间：
2016-12-18
期刊：
影响因子：
--
通讯作者：
Franks AE
中科院分区：
文献类型：
--
作者：
Bereza-Malcolm L;Aracic S;Franks AE
在过去几十年中，铅在制造业中的使用量大幅减少。然而，以前含铅产品的广泛使用及其不正确的处置已导致环境污染。有害量的铅通过吸入、摄入或直接接触积累，对所有生物体构成威胁，导致铅中毒。本研究利用合成生物学原理研制了一种基于质粒的全细胞细菌生物传感器，用于铅的检测。该先导生物传感器构建体的遗传元件由编码调节蛋白的pbrR及其趋异启动子区和无启动子的gfp组成。GFP表达受PbrR控制，以响应铅的存在。将先导生物传感器遗传元件克隆到低拷贝数宽宿主范围质粒上，该质粒可以稳定存在于一系列实验室和环境分离株中，包括假单胞菌属、希瓦氏菌属和肠杆菌属。构建的生物传感器被认为是敏感的，快速的，和具体的，因此，可以作为监测工具，铅污染的水。
The use of lead in manufacturing has decreased significantly over the last few decades. However, previous widespread use of lead-containing products and their incorrect disposal has resulted in environmental contamination. Accumulation of harmful quantities of lead pose a threat to all living organisms, through inhalation, ingestion, or direct contact, resulting in lead poisoning. This study utilized synthetic biology principles to develop plasmid-based whole-cell bacterial biosensors for detection of lead. The genetic element of the lead biosensor construct consists of pbrR, which encodes the regulatory protein, together with its divergent promoter region and a promoterless gfp. GFP expression is controlled by PbrR in response to the presence of lead. The lead biosensor genetic element was cloned onto a low-copy number broad host range plasmid, which can stably exist in a range of laboratory and environmental isolates, including Pseudomonas, Shewanella, and Enterobacter. The biosensors constructed were found to be sensitive, rapid, and specific and could, as such, serve as monitoring tools for lead-contaminated water.