采用SELDI蛋白质芯片技术研究氧化亚铁硫杆菌对磷酸盐缺失的反应(英文)

Research on The Proteome Response of Acidithiobacillus ferrooxidans to Phosphate Starvation by SELDI-Protein Chip Technologies

氧化亚铁硫杆菌(At.f)是能够利用Fe2+和硫化矿来获取能量的一种化能自养菌.这种细菌在金属硫化矿的生物浸出中起着重要的作用.在硫化矿的生物浸出过程中,浸矿细菌通常会遇到多种胁迫条件,如温度的变化、营养成分的缺失和pH值的变化等,这些因素会影响到细菌的活性.因此对在胁迫条件下这类细菌的应急反应生理机制的研究具有重要的意义.SELDI蛋白质芯片技术是近年一种高通量的蛋白质组学研究技术.测定了以Fe2+为能源正常条件培养的At.f和磷酸盐缺失培养At.f的生长情况,绘制了相应的生长曲线;采用NP20蛋白质芯片,对At.f总蛋白的蛋白质芯片上样量进行了优化.在此基础上,采用IMAC-Cu、SAX2、WCX2三种特异性SELDI蛋白质芯片技术,获取了磷酸盐缺失培养At.f与正常条件培养的At.f的比较蛋白质图谱,采用软件对比较蛋白质图谱进行分析,发现了磷酸盐缺失培养At.f的13个明显差异表达的蛋白质分子,为进一步分离鉴定这些差异表达蛋白质奠定了基础.

A cidithiobacillus ferrooxidans is a chemolithotrophic microorganism capable of using ferrous ions and sulphides as energy sources. This microorganism has an important role in the bioleaching of minerals. During this process, the bacteria arc normally subjected to several stressing conditions, such as temperature changes, lack of nutrients or pH changes, which may affect the efficiency of the bacterial action. SELDI is a recent technology that allows for high-throughput proteomics studies. The Protein Chip SELDI technology was used to generate comparative protein profiles of A cidithiobacillus ferrooxidans grown under phosphate starvation or normal condition additionally adding Fe^2＋ as energy resource. There were 13 significantly differential expressed protein＇s peaks found by using SELDI Protein Chip technologies, which made a solid foundation for further isolation these low molecular proteins by adopting technologies such as HPLC etc.