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Application of Firefly Luciferase (Luc) as a Reporter Gene for the Chemoautotrophic and Acidophilic Acidithiobacillus spp.

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Abstract

Acidithiobacillus spp. are the most active bacteria in bioleaching and bioremediation, because of their remarkable extreme environmental adaptabilities and unique metabolic characteristics. The researches on regulatory mechanisms of energy metabolism and stress resistance are critical for the understanding and application of Acidithiobacillus spp. However, the lack of an ideal reporter gene has become an obstacle for studying genes expression and regulatory mechanism in these chemoautotrophic bacteria. In this study, we reported the firefly luciferase as a reporter gene for Acidithiobacillus caldus (A. caldus) and created a firefly luciferase (Luc) reporter system. The Luc system was applied for the quantitative analysis of the transcription strength of the promoters of tetH gene and the feoA gene in A. caldus. Moreover, the regulating effect of ferric uptake regulator (Fur) on the feoP gene in A. caldus was determined using the Luc system. The Luc reporter system is not only used in the study of regulatory mechanism of A. caldus, but also applied in the researches of other Acidithiobacillus species. Therefore, this study provides a new useful tool for the studies on the molecular biological mechanism and synthetic biological modification of these chemoautotrophic bacteria, which would promote the industrial application of Acidithiobacillus spp.

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Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (31570036, 31872621, 31570041.), the Project of Taishan Industry Leading Talent in Shandong province (LJNY201603), the State Key Laboratory of Microbial Technology Foundation (M2017-01), People’s Republic of China. We are grateful to Pro. Jun Fu from Shandong University for providing the plasmid pBR322-Luc containing firefly luciferase gene. We thank the supports from Core Facilities Sharing Platform for Life and Environment Sciences of Shandong University, including Rui Wang for luciferase activity detection, Cheng-jia Zhang and Nan-nan Dong for providing bacteriological incubator.

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Authors and Affiliations

  1. State Key Laboratory of Microbial Technology, Shandong University, No. 72 Binhai Road, Qingdao, 266237, People’s Republic of China

    Xianke Chen, Xiujie Liu, Yuhui Gao, Jianqiang Lin, Xiangmei Liu, Xin Pang, Jianqun Lin & Linxu Chen

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  1. Xianke Chen
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  2. Xiujie Liu
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  3. Yuhui Gao
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  7. Jianqun Lin
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Contributions

XC contributes to the performance of experiments, the analysis of the data, the writing and visualization of the manuscript. XL and YG contribute to methods investigation and data validation. JL, XL and XP provide foundings and suggestions for this manuscript. JL and LC contribute to the design of experiment, the organization and modification of the manuscript.

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Correspondence to Jianqun Lin or Linxu Chen.

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Chen, X., Liu, X., Gao, Y. et al. Application of Firefly Luciferase (Luc) as a Reporter Gene for the Chemoautotrophic and Acidophilic Acidithiobacillus spp.. Curr Microbiol 77, 3724–3730 (2020). https://doi.org/10.1007/s00284-020-02195-w

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