摘要
目的:旨在探索和分析嗜酸氧化亚铁硫杆菌重金属抗性基因Cd(II)-Responsive Transcriptional Regulator(CmtR)的解毒机制。方法:实验首先通过生物信息学的方法分析发现,嗜酸氧化亚铁硫杆菌一个重金属抗性基因(CmtR)序列上含有多个半胱氨酸的编码基因。实验将含有抗性基因CmtR的重组质粒转入E.Coli DH5α中,验证正确后,加IPTG诱导表达,分析了细胞内Cys、GSH和重金属含量的变化。结果:SDS-PAGE分析表明,CmtR蛋白质分子量大小约为19 kDa;抗性基因CmtR的解毒机制可能是由于Cys、GSH能够特异性结合重金属离子并将其排出体外,从而达到解毒的目的。结论:初步确定了嗜酸氧化亚铁硫杆菌重金属抗性基因CmtR的解毒机制。
Objective: To identify and analyze the gene of CmtR in the heavy-metal detoxify mechanism in Acidithiobacillus ferrooxidans ATCC 23270. Methods: This study analyzes that there were multiple encoding Cysteine genes Cd(Ⅱ)-responsive transcriptional regulator( CmtR) in A. ferrooxidans ATCC23270 by using bioinformatics firstly. We transfered the recombinational plasmid containing CmtR into E.Coli DH5α,after being verified correct,adding IPTG to induce CmtR overexpressing,and then analyzed the content change of intracellular Cys,GSH,and the related heavy metal. Results: SDS-PAGE analysis suggested that the size of protein is about 19 KD,the detoxification mechanisms of CmtR may was that Cysand GSH could specificly combine with heavy metal ions and transport them out of the organisms.Conclusion: the probable detoxification mechanisms of CmtR in A. ferrooxidans ATCC 23270 was confirmed preliminarily.
出处
《科技通报》
2018年第5期59-65,共7页
Bulletin of Science and Technology
基金
国家自然科学基金项目(51264029,41561094)
内蒙古自治区青年科技英才计划项目(NJYT-14-B12)
内蒙古自治区应用技术研究与开发资金项目
内蒙古科技大学创新基金(2014QNGG05)
