大肠杆菌O157：H7narX与narP缺失株的构建及其对细胞粘附与A／E损伤的观察  

Construction of narX and narP mutants and their effect on adhesion and A/E lesion in E. coli O157:H7

作　　者：徐雪芳[1] 刘学通[1] 熊衍文[1] 徐建国[1] 

机构地区：[1]中国疾病预防控制中心传染病预防控制所,传染病预防控制国家重点实验室,北京102206

出　　处：《中国人兽共患病学报》2013年第4期330-334,共5页Chinese Journal of Zoonoses

基　　金：传染病预防控制国家重点实验室留学归国人员启动经费项目(2012SKLID401);国家“艾滋病和病毒性肝炎等重大传染病防治”科技重大专项(2011ZX10004-001)资助~~

摘　　要：目的大肠杆菌O157∶H7是一种重要的可以引起人类严重疾病的肠出血性大肠杆菌(enterohaemorrhagic E.coli,EHEC)血清型。为获得narX与narP缺失株并观察其对细胞粘附与A/E(attaching and effacing)损伤的影响。方法本研究采用λRed重组技术,利用PCR产物,通过Bet和Exo蛋白促进同位交换的方法,构建参与呼吸系统调控的narX和narP突变株。结果成功构建了narX和narP突变株,通过细胞粘附实验和免疫荧光染色,发现两个突变株对EBL细胞的粘附力都有明显下降,而在A/E损伤形成能力方面,只有narX突变株有明显下降。结论 narX和narP均对细胞的粘附有影响,narX对细胞A/E损伤有影响。进一步的机制还需深入的实验研究来证明。E. coli O157 ： H7 is one of the most important serotypes of EHEC causing diseases in human. To obtain narX and narP mutants and investigate their effect on adhesion and A/E lesion in E. coli O157, narX and narP mutants were constructed by ）,-Red recombineering technology promoted by Bet and Exo proteins using PCR products. By using bacterial in- fection assays and immunofluorescence microscopy, it was found that the adhesion ability were decreased both in narX and narP mutant. In addition, the A/E lesion formation was only down-regulated in narX mutant. In conclusion, narX and narP gene might impact the adhesion and narX could influence A/E lesion formation.

关 键 词：大肠杆菌O157 H7 narX NARP 

分 类 号：Q933[生物学—微生物学]