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Copy number of ArsR reporter plasmid determines its arsenite response and metal specificity

  • Environmental biotechnology
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Abstract

The key component in bacteria-based biosensors is a transcriptional reporter employed to monitor induction or repression of a reporter gene corresponding to environmental change. In this study, we made a series of reporters in order to achieve highly sensitive detection of arsenite. From these reporters, two biosensors were developed by transformation of Escherichia coli DH5α with pLHPars9 and pLLPars9, consisting of either a high or low copy number plasmid, along with common elements of ArsR-luciferase fusion and addition of two binding sequences, one each from E. coli and Acidithiobacillus ferrooxidans chromosome, in front of the R773 ArsR operon. Both of them were highly sensitive to arsenite, with a low detection limit of 0.04 μM arsenite (~ 5 μg/L). They showed a wide dynamic range of detection up to 50 μM using high copy number pLHPars9 and 100 μM using low copy number pLLPars9. Significantly, they differ in metal specificity, pLLPars9 more specific to arsenite, while pLHPars9 to both arsenite and antimonite. The only difference between pLHPars9 and pLLPars9 is their copy numbers of plasmid and corresponding ratios of ArsR to its binding promoter/operator sequence. Therefore, we propose a working model in which DNA bound-ArsR is different from its free form in metal specificity.

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Funding

This work was supported by the High-level Leading Talent Introduction Program of GDAS (2016GDASRC-0208) and the Science and Technology Planning Project of Guangzhou City (201707020021) to XL, the National Natural Science Foundation of China (31600077) and the China Postdoctoral Science Foundation Grant (2017M612622) to YF, and the Science and Technology Project of Guangdong Province (2016B070701017) to MX.

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Author notes

  1. Xianqiang Li

    Present address: Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, 100 Central Xianlie Road, Guangzhou, 510070, Guangdong, China

  2. Yun Fang and Chunjie Zhu have equal contribution to the study.

Authors and Affiliations

  1. Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, 100 Central Xianlie Road, Guangzhou, 510070, Guangdong, China

    Yun Fang, Chunjie Zhu, Xingjuan Chen, Meiying Xu, Guoping Sun & Jun Guo

  2. State Key Laboratory of Applied Microbiology Southern China, Guangzhou, China

    Yun Fang, Chunjie Zhu, Xingjuan Chen, Meiying Xu, Guoping Sun, Jun Guo & Xianqiang Li

  3. Science and Technology Library of Guangdong Province, Guangdong Institute of Science and Technology Information and Development Strategy, Guangzhou, China

    Yan Wang

  4. Signosis Inc., 1700 Wyatt Drive, suite10-12, Santa Clara, CA, USA

    Jinnon Yoo, Cuijuan Tie, Xin Jiang & Xianqiang Li

Authors
  1. Yun Fang
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  2. Chunjie Zhu
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Correspondence to Meiying Xu or Xianqiang Li.

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Fang, Y., Zhu, C., Chen, X. et al. Copy number of ArsR reporter plasmid determines its arsenite response and metal specificity. Appl Microbiol Biotechnol 102, 5753–5761 (2018). https://doi.org/10.1007/s00253-018-9042-1

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  • DOI: https://doi.org/10.1007/s00253-018-9042-1

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