阪崎肠杆菌蛋白质组学研究及特征蛋白的质谱分析  

Two-dimensional Electrophoresis and MALDITOF/MS Analysis for Enterobacter sakazakii Specific Protein

作　　者：杨捷琳 魏黎明[2] 顾鸣 方晓明 韩伟 

机构地区：[1]上海市出入境检验检疫局,上海200135 [2]复旦大学生物医学研究院,上海200032

出　　处：《复旦学报（自然科学版）》2008年第5期620-626,632,共8页Journal of Fudan University：Natural Science

基　　金：国家质量监督检验检疫总局科技项目(2006IK108)

摘　　要：阪崎肠杆菌为婴幼儿奶粉中新出现的食源性致病菌,生态学及毒力因子尚不清楚.利用双向电泳通过裂解液组成、上样量、聚焦时间等相关技术的比较研究和条件优化,获得阪崎肠杆菌菌体全蛋白双向电泳图谱,提取部分蛋白质点进行胰酶酶解,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF/MS)测定肽质量指纹图谱,MASCOT软件查询Swiss-Prot数据库,最终鉴定出阪崎肠杆菌中特异性蛋白,包括大肠杆菌氧化态硫醇过氧化物酶A链,大肠杆菌SfaA蛋白,麦芽糖高亲和结合蛋白A链,饥饿应激蛋白,阪崎肠杆菌组成型表达蛋白,鼠伤寒沙门氏菌脱氧核糖磷酸醛缩酶,大肠杆菌O157铁离子摄取调节子等,反映出阪崎肠杆菌与肠杆菌科近似菌株蛋白表达的相似性及生长特性.Powdered infant formula pathogen infection by Enterobacter sakazakii is closely correlated with meningitis, severe neurological sequelae such as hydrocephlus, quadraplegia. Although considerable progress has been made during recent years to establish the bacterium detection method, little have been known about the mechanism of pathogenesis of E. sakazakii infection. The proteomic techniques were applied in this work to globally analyze the protein expression profiles of E. sakazakii, aiming at elucidating the components of E. sakazakii replication and the cellular function at stationary growth station. The protein mixtures of three subeellular fractions from E. sakazakii were separated by 2-DE under pH 3-10 gradients. Specifically expressed spots were identified by MALDI-TOF MS, followed by database searching. A total of 50 comparative proteins were identified unambiguously, including proteins associated with host eytoskeleton, intracellular traffic, oxidative and translation, proliferation, etc.

关 键 词：阪崎肠杆菌 双向电泳 质谱 特征性蛋白 

分 类 号：Q936[生物学—微生物学]