用于检测生物可利用铜的恶臭假单胞菌全细胞生物传感器的构建与优化
Construction and optimization of a Pseudomonas putida whole-cell bioreporter for detection of bioavailable copper.
作者信息
Li Peng-Song, Peng Zhi-Wen, Su Jie, Tao Hu-Chun
机构信息
Key Laboratory for Heavy Metal Pollution Control and Reutilization, School of Environment and Energy, Peking University Shenzhen Graduate School, Shenzhen, 518055, China,
出版信息
Biotechnol Lett. 2014 Apr;36(4):761-6. doi: 10.1007/s10529-013-1420-2. Epub 2013 Dec 10.
A Pseudomonas putida whole-cell bioreporter for detecting bioavailable copper was constructed by inserting a CueR-regulated sensor element upstream of a promoterless green fluorescent protein (GFP) reporter gene. The constructed bioreporter cells expressed GFP only in response to Cu and Ag when cultivated in different metal salt solutions. M9 supplemented medium provided higher sensitivity compared with LB medium. The optimal test condition was cell suspension with an OD600 of 0.4-0.5 incubated at 30 °C. The detection range of Cu is 1-70 mg/l under optimal test condition in M9 supplemented medium.
通过将一个受CueR调控的传感器元件插入无启动子绿色荧光蛋白（GFP）报告基因的上游，构建了一种用于检测生物可利用铜的恶臭假单胞菌全细胞生物传感器。当在不同金属盐溶液中培养时，构建的生物传感器细胞仅在对铜和银作出反应时表达绿色荧光蛋白。与LB培养基相比，添加M9的培养基具有更高的灵敏度。最佳测试条件是在30℃下孵育OD600为0.4-0.5的细胞悬液。在添加M9的培养基中，在最佳测试条件下铜的检测范围为1-70mg/l。
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