铜绿假单胞菌外膜亚蛋白质组图谱的建立  

Establishment of Subproteome Map of Pseudomonas aeruginosa OMPs

作　　者：章喻军[1] 王三英[1] 

机构地区：[1]厦门大学生命科学学院,福建厦门361005

出　　处：《厦门大学学报（自然科学版）》2008年第1期111-115,共5页Journal of Xiamen University：Natural Science

基　　金：国家自然科学基金973项目(2006CB101807)资助

摘　　要：铜绿假单胞菌是人类重要的病原菌,容易造成免疫缺陷患者如烧伤、艾滋病、癌症或囊性纤维化感染,对该菌外膜蛋白的研究有助于了解该菌的致病机制,为进一步疫苗研制奠定基础.采用月桂酰基氨酸钠的方法提取了铜绿假单胞菌的外膜蛋白,首次建立了外膜亚蛋白质组学表达图谱,鉴定了23个外膜蛋白,占所鉴定蛋白的82.1%.然后对其中的一个外膜蛋白基因PA3988进行了克隆及原核表达与纯化,经免疫小鼠获得抗血清后对外膜蛋白进行双向Westernblotting分析,验证了质谱的结果.这些研究结果对铜绿假单胞菌外膜蛋白的深入研究具有积极作用.Pseudomonas aeruginosa is an opportunistic human pathogen,especially for immunocompromised individuals such as the patients with burn victims,AIDS,cancer or cystic fibrosis.Meanwhile,bacterial outer membrane proteins（OMPs） are important for Gram-negative bacteria because they are involved in adhesion to and invasion of host cells,and in response to the chemical and physical factors of the external milieu.In the current study,the OMPs of P.aeruginosa were investigated with the use of sub-proteomic methodologies by analysis of sodium lauroyl sarcosine-insoluble fraction.Thirty-one proteins were identified,of which 23 were OMPs,accounting for 82.1%.Their function was assigned.Furthermore,PA3988 gene,one of the identified 23 protein genes,was randomly selected for cloning.The cloned gene was expressed for antibody preparing.The prepared antibody was used as the primary antibody in 2-DE Western blotting for confirmation of an identified spot by mass spectrometry.This established complete proteomic profile sheds new insight into the function（s） of E.coli outer envelope.

关 键 词：外膜蛋白基因 铜绿假单胞菌 蛋白质组图谱 蛋白质组学 免疫缺陷 致病机制 月桂酰基 疫苗研制 

分 类 号：S858.235[农业科学—临床兽医学] Q939.112[农业科学—兽医学]