摘要
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.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2008年第11期967-971,共5页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金资助项目（30470096）
