氧化亚铁硫杆菌中铜代谢相关两个基因的差异(英文)  

Differences between Two Copper Related Genes in the Genome of Acidithiobacillus ferrooxidans

作　　者：罗焱杰[1] 张成桂[1] 刘元东[1] 吴学玲[1] 管昊[1] 廖荷欢[1] 罗海浪[1] 刘学端[1] 邱冠周[1] 

机构地区：[1]中南大学资源加工与生物工程学院,湖南长沙410083

出　　处：《生物技术》2009年第2期9-13,共5页Biotechnology

基　　金：国家"973"项目(No.2004CB619201);国家自然科学基金项目(No.30428014,30670044,50621063)资助

摘　　要：目的:氧化亚铁硫杆菌(Acidithiobacillus ferrooxidans)在微生物冶金中发挥着重要的作用,研究其铜代谢机理有着十分典型的意义。在A.ferrooxidans全基因组序列数据库中,4个基因被注释与铜代谢相关。其中两个基因,Afe0454和Afe1073目前为止未发现有实验报道。本文旨在研究Afe0454和Afe1073与铜代谢的相关性。方法:通过一系列的方法如实时定量PCR、反转录PCR、序列分析,将基因导入抗铜基因缺陷的大肠杆菌(Escherichia coli)菌株中等,研究了Afe0454和Afe1073。结果:与Afe0454相比,Afe1073的表达对铜压力更敏感;Afe1073作为一个转录子单独转录,而Afe0454与Afe0453一起转录;序列分析显示Afe1073表达一种典型的重金属离子泵P1b1型ATP酶,而Afe0454表达一种未知功能的跨膜蛋白;不像Afe0454,Afe1073能使P1b型ATP酶敲除的大肠杆菌菌株铜抗性提高。结论:单独转录的Afe1073比Afe0454在铜代谢中发挥的作用更加明显。Objective：Acidithiobacillus ferrooxidans plays a very important role in bioleaching industry, and its mechanism of copper homeostasis is very typical. In on-line A.ferrooxidans sequence database （http：www.tigr.org）, four genes were annotated to be involved in copper homeostasis, until now no experimental research about two of them, Afe0454 and Afe1073 has been reported. This paper aimed to identify the role gene Afe1073 and Afe0454 playing in the copper homeostasis. Method：Real-time PCR, Reverse transcriptase-PCR, sequence analysis and complementation experiment to copper sensitive strain of Escherichia coli were carried out to identify the gene Afe1073 and Afe0454. Result：Compared with Afe0454, Afe1073 is more intensive response to copper stress, unlike the transcripton of Afe1073 containing only Afe1073, gene Afe0454 cotranscribes with Afe0453. In addition, based on sequence analysis, gene Afe1073 expresses a P1b1-type ATPase which is a kind of heavy metal pump, but the protein expressed by Afe0454 is just a kind of unknown functional transmembrane protein. Moreover, plasmid carrying Afe1073 can partially rescue the P1b-type ATPase disrupted Escherichia coli, but Afe0454 cannot do so. Conclusion：All of these suggest the role of single transcribed gene Afe1073 playing in copper homeostasis is more obvious and apparent compared with Afe0454.

关 键 词：氧化亚铁硫杆菌 铜代谢 P1b型ATP酶 实时定量PCR 

分 类 号：Q751[生物学—分子生物学]