CAJ | 학술논문

为了阐述 O157:H7与 O157的致病力差异与蛋白质表达差异之间的关系,我们利用双向电泳技术和质谱技术对大肠杆菌两类菌株之间进行蛋白质组学差异分析.菌体蛋白经双向电泳技术分离后,利用软件Image MasterTM 2D Platinum 7.0对所得双向电泳图谱进行差异分析并借助质谱技术对差异蛋白质点进行鉴定,获得了背景清晰、分辨率高、重复性好的菌体总蛋白的双向电泳图谱.两样品图谱差异分析结果表明,共确定了28个有效的蛋白质差异点(Av. ratio>2.0, ANOVA<0.05),经质谱鉴定将这28个差异点注释成23种蛋白质.对得到注释的23种蛋白质进行功能分类,发现7类蛋白质与致病性密切相关,即溶菌酶抑制剂、通用应激蛋白、LuxS、鞭毛蛋白以及 3种外膜蛋白.本结果将为进一步研究 O157:H7菌株的致病因子,探究O157:H7与其它大肠杆菌菌株的致病力差异,以及建立快速鉴别诊断 O157:H7的试剂盒提供重要的依据.
위료천술 O157:H7여 O157적치병력차이여단백질표체차이지간적관계,아문이용쌍향전영기술화질보기술대대장간균량류균주지간진행단백질조학차이분석.균체단백경쌍향전영기술분리후,이용연건Image MasterTM 2D Platinum 7.0대소득쌍향전영도보진행차이분석병차조질보기술대차이단백질점진행감정,획득료배경청석、분변솔고、중복성호적균체총단백적쌍향전영도보.량양품도보차이분석결과표명,공학정료28개유효적단백질차이점(Av. ratio>2.0, ANOVA<0.05),경질보감정장저28개차이점주석성23충단백질.대득도주석적23충단백질진행공능분류,발현7류단백질여치병성밀절상관,즉용균매억제제、통용응격단백、LuxS、편모단백이급 3충외막단백.본결과장위진일보연구 O157:H7균주적치병인자,탐구O157:H7여기타대장간균균주적치병력차이,이급건립쾌속감별진단 O157:H7적시제합제공중요적의거.
Differential proteomic analysis between E. coli strains O157:H7 and O157 was carried out, in order to clarify the relationship between virulence and differentially expressed proteins of different E. coli strains. Two-dimensional electrophoresis (2-DE) was used to separate bacterial proteins and Image MasterTM 2D Platinum 7.0 software performed variation analysis of the obtained 2D maps. The differentially expressed proteins were identified by mass spectrometry. The 2D maps of bacterial total protein with clear background, high resolution and good reproducibility were obtained. The variation analysis of two samples was performed and a total of 28 differentially expressed protein spots (by fliter Av. ratio>2.0 and ANOVA<0.05) were detected. These protein spots were identified as 23 unique protein groups by mass spectrometry. The functional classification of annotated proteins indicated that seven kinds of proteins were closely related to bacterial virulence including C-lysozyme inhibitor, universal stress protein, LuxS, flagellin and 3 outer membrane proteins. This research provides an important proteomic basis for studying virulence factors, finding pathogenicity differences and establishing the rapid diagnosis kit of the bacterial strain O157:H7.