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Construction and optimization of a Pseudomonas putida whole-cell bioreporter for detection of bioavailable copper

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Abstract

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.

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Acknowledgments

This research was supported by the Shenzhen municipal government under grants of CXB201111240110A and JCYJ20130329174424934.

We thank Prof. Steven Lindow of University of California, Berkeley for providing the plasmid pPROBE-NT and Priscilla Young for English-language editing.

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Authors and Affiliations

  1. Key Laboratory for Heavy Metal Pollution Control and Reutilization, School of Environment and Energy, Peking University Shenzhen Graduate School, Shenzhen, 518055, China

    Peng-Song Li, Zhi-Wen Peng, Jie Su & Hu-Chun Tao

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  1. Peng-Song Li
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  2. Zhi-Wen Peng
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  3. Jie Su
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  4. Hu-Chun Tao
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Correspondence to Hu-Chun Tao.

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Li, PS., Peng, ZW., Su, J. et al. Construction and optimization of a Pseudomonas putida whole-cell bioreporter for detection of bioavailable copper. Biotechnol Lett 36, 761–766 (2014). https://doi.org/10.1007/s10529-013-1420-2

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  • DOI: https://doi.org/10.1007/s10529-013-1420-2

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