生物技术
生物技術
생물기술
BIOTECHNOLOGY
2009年
2期
9-13
,共5页
罗焱杰%张成桂%刘元东%吴学玲%管昊%廖荷欢%罗海浪%刘学端%邱冠周
囉焱傑%張成桂%劉元東%吳學玲%管昊%廖荷歡%囉海浪%劉學耑%邱冠週
라염걸%장성계%류원동%오학령%관호%료하환%라해랑%류학단%구관주
氧化亚铁硫杆菌%铜代谢%P1b型ATP酶%实时定量PER
氧化亞鐵硫桿菌%銅代謝%P1b型ATP酶%實時定量PER
양화아철류간균%동대사%P1b형ATP매%실시정량PER
Acidithiobacillus ferrooxidans%copper homeostasis%Plb- type ATPase%Real- time PCR
目的:氧化亚铁硫杆菌(Acidithiobocllius 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PER、反转录PER、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.
目的:氧化亞鐵硫桿菌(Acidithiobocllius ferrooxidans)在微生物冶金中髮揮著重要的作用,研究其銅代謝機理有著十分典型的意義.在A.ferrooxidans全基因組序列數據庫中,4箇基因被註釋與銅代謝相關.其中兩箇基因,Afe0454和Afe1073目前為止未髮現有實驗報道.本文旨在研究Afe0454和Afe1073與銅代謝的相關性.方法:通過一繫列的方法如實時定量PER、反轉錄PER、序列分析,將基因導入抗銅基因缺陷的大腸桿菌(Escherichia coli)菌株中等,研究瞭Afe0454和Afe1073.結果:與Afe0454相比,Afe1073的錶達對銅壓力更敏感;Afe1073作為一箇轉錄子單獨轉錄,而Afe0454與Afe0453一起轉錄;序列分析顯示Afe1073錶達一種典型的重金屬離子泵P1b1型ATP酶,而Afe0454錶達一種未知功能的跨膜蛋白;不像Afe0454,Afe1073能使P1b型ATP酶敲除的大腸桿菌菌株銅抗性提高.結論:單獨轉錄的Afe1073比Afe0454在銅代謝中髮揮的作用更加明顯.
목적:양화아철류간균(Acidithiobocllius ferrooxidans)재미생물야금중발휘착중요적작용,연구기동대사궤리유착십분전형적의의.재A.ferrooxidans전기인조서렬수거고중,4개기인피주석여동대사상관.기중량개기인,Afe0454화Afe1073목전위지미발현유실험보도.본문지재연구Afe0454화Afe1073여동대사적상관성.방법:통과일계렬적방법여실시정량PER、반전록PER、서렬분석,장기인도입항동기인결함적대장간균(Escherichia coli)균주중등,연구료Afe0454화Afe1073.결과:여Afe0454상비,Afe1073적표체대동압력경민감;Afe1073작위일개전록자단독전록,이Afe0454여Afe0453일기전록;서렬분석현시Afe1073표체일충전형적중금속리자빙P1b1형ATP매,이Afe0454표체일충미지공능적과막단백;불상Afe0454,Afe1073능사P1b형ATP매고제적대장간균균주동항성제고.결론:단독전록적Afe1073비Afe0454재동대사중발휘적작용경가명현.
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 ho-meostasis, 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 oomplementation experiment to copper sensitive strain of Escheriehia coli were carried out to identify the gene Aft 1073 and Afe0454. Result: Com-pared with Afe0454, Afe1073 is more intensive response to copper stress; unlike the transcripton of Afe1073 containing only Afe1073, gene Afe0454 cotranseribes 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 Af0454 is just a kind of unknown functional transmembrane protein. Moreover, plasmid carrying Afe1073 can partially rescue the P1b- type ATPase disrupted Escherichia coil; 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.
