Application of Firefly Luciferase (Luc) as a Reporter Gene for the Chemoautotrophic and Acidophilic Acidithiobacillus sp
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Application of Firefly Luciferase (Luc) as a Reporter Gene for the Chemoautotrophic and Acidophilic Acidithiobacillus spp. Xianke Chen1 · Xiujie Liu1 · Yuhui Gao1 · Jianqiang Lin1 · Xiangmei Liu1 · Xin Pang1 · Jianqun Lin1 · Linxu Chen1 Received: 17 February 2020 / Accepted: 3 September 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
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.
Introduction The γ-proteobacterium Acidithiobacillus spp., distributed in acidic sulfur/iron-containing habitats (acid mine drainages and hot springs) [1, 2], can oxidize various reduced inorganic sulfur compounds to fix CO2 for autotrophic growth (some species also have the ferrous oxidation capacity) [3]. Due to the highly efficient sulfur/iron metabolism and strong resistance mechanisms (extremely acidic, highly concentrations of metals, etc.), Acidithiobacillus spp. have been widely used in bioleaching and bioremediation [1, 4–6], such as recovering valuable metal copper from low-grade sulfidic ores or acid mine drainage (AMD) [5], bioleaching treatment waste electrical and electronic equipment (WEEE) [6]. Thus, research on the regulatory mechanisms of genes involved in sulfur/iron metabolism and stress resistance, are * Jianqun Lin [email protected] * Linxu Chen [email protected] 1
State Key Laboratory of Microbial Technology, Shandong University, No. 72 Binhai Road, Qingdao 266237, People’s Republic of China
of significance for revealing the survival strategies of these acidophilic bacteria in the extremely environments, and promoting the synthetic biology modification of Acidithiobacillus spp. for bioleaching and bioremediation. A variety of reporter genes, such as lacZ, gusA, and gfp, ha
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