目的研究致肾盂。肾炎大肠杆菌(UPEC)132与UPEC J96及非致病性大肠杆菌K-12MG1655的蛋白质组表达差异。方法采用双向凝胶电泳技术(2-DE)比较UPEC 132、UPEC J96与非致病性大肠杆菌K-12MG1655的菌体蛋白表达图谱差异,差异蛋白用胰蛋白酶进行胶内酶切,肽混合物使用基质辅助激光解吸-电离飞行时间质谱仪(MALDI—TOF—MS)进行质谱分析,将肽质量指纹谱数据输入互联网上的蛋白质数据库进行检索。结果UPEC132识别蛋白点数(466±11)明显高于Ecoli K-12 MG1655(338±15),也高于UPEC J96(382±12);3菌株共有的蛋白点为298个,2株UPEC共有的蛋白点为56个,UPEC 132特有的蛋白点为89个。MALDI—TOF—MS分析获得UPEC132特异的及显著上调表达的蛋白点22个,涉及毒力因子、物质代谢转运、蛋白合成调节、生物氧化及未知功能的多种蛋白质。结论UPEC菌株与非致病性大肠杆菌间的蛋白质组存在差异,UPEC 132与J96蛋白质组间也有显著不同,为其致病机制的深入研究提供线索。
Objective To study proteome variation between uropathogenic E. coli (UPEC)132, UPEC J96 and non-uropathogenic E. coli K-12 MG1655. Methods Two-dimensional gel electrophoresis(2- DE ) was applied to compare the differential expression proteins between UPEC 132, UPEC J96 and non-uropathogenic E. coli K-12 MG1655. The differential expression proteins were digested in gel by enzyme. The mass of generated peptides were measured by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). The data obtained from peptide mass fingerprinting (PMF) were researched using the internet available database. Results The number of protein spots recognized from UPEC 132 was 466± 11, significantly more than that of E. coil K-12 MG1655 (338 ± 15) and UPEC J96(382 ± 12) ; there were 298 protein spots shared by the three E. coli strains, 56 protein spots shared by two UPEC strains, and 89 protein spots characterized by UPEC 132. Twenty-two differential expression or significantly increased expression protein spots, involved in virulence factors, metabolism and transportation, regulation of protein synthesis, biological oxidation and unknown functions, were successfully identified by MALDI- TOF-MS. Conclusion The proteome from UPEC 132 and non-uropathogenic E. coli K-12 MG1655, or UPEC 132 and UPEC J96 was differentially expressed. It will provide important information on the pathogenesis of UPEC 132.