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在大肠杆菌中克隆、表达和生物信息学分析嗜酸氧化亚铁硫杆菌 L1 中的重金属抗性基因 afe_1862。

Cloning, Expression, and Bioinformatics Analysis of Heavy Metal Resistance Gene afe_1862 from Acidithiobacillus ferrooxidans L1 in Escherichia coli.

机构信息

School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou, 730050, China.

Lanzhou Institute of Husbandry and Pharmaceutical Science of CAAS, Lanzhou, 730050, China.

出版信息

Biol Trace Elem Res. 2019 May;189(1):291-300. doi: 10.1007/s12011-018-1462-3. Epub 2018 Aug 16.

Abstract

Molecular studies of copper and cadmium resistances in acidophilic bacteria are significant in biomining. In this study, afe_1862, which encodes a heavy metal-binding protein in Acidithiobacillus ferrooxidans L1, was amplified using PCR, cloned into the pET32a plasmid, and sequenced. Following SDS-PAGE analysis, optimization of the expression conditions and heterologous overexpression of afe_1862 in Escherichia coli BL21 in the presence of Cu and Cd were studied as well. The results indicated that AFE_1862 has higher resistance to Cu than Cd. Bioinformatics analysis illustrated that AFE_1862 has a conserved HMA domain containing heavy metal-binding sites, which may play a role in transporting or detoxifying heavy metals.

摘要

在生物采矿中,研究嗜酸菌的铜和镉抗性的分子机制具有重要意义。在这项研究中,使用 PCR 扩增了嗜酸氧化亚铁硫杆菌 L1 中的编码重金属结合蛋白的 afe_1862,将其克隆到 pET32a 质粒中,并进行测序。通过 SDS-PAGE 分析,研究了在存在 Cu 和 Cd 的情况下,afe_1862 在大肠杆菌 BL21 中的表达条件优化和异源过表达。结果表明,AFE_1862 对 Cu 的抗性高于 Cd。生物信息学分析表明,AFE_1862 具有保守的 HMA 结构域,包含重金属结合位点,可能在运输或解毒重金属方面发挥作用。

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