军事医学2017，Vol.41Issue(12)：978-982,5.DOI:10.7644/j.issn.1674-9960.2017.12.007
肠出血性大肠杆菌z5150毒素基因在细胞黏附过程中的作用
Role of enterohemorrhagic E.coli toxin gene z5150 in cell adhesion
摘要
Abstract
Objective To construct EHEC△z5150 using Red recombination and explore its pathogenicity.Methods Overexpression of the z5150 gene was used to test its toxicity.The kanamycin resistant gene flanked by homologues of target genes was amplified by PCR before being transferred into EHEC containing pKD 46 plasmid,and the target fragments were recombined into genome by Arabia sugar.The kanamycin resistance gene was removed by pFLP 2 plasmid mediated FLP site-specific recombination.Cell adhesion ability and virulence of the mutant were detected by infecting HT 29 cells and BALB/c female mice,respectively.Results and Conclusion The mutant strain EHEC △z5150 was successfully constructed.The absence of z5150 did not affect the normal growth of bacteria,but after excessive expression,it inhibited the growth.Deletion of z5150 showed significant difference in intestinal epithelial cell adhesion and pathogenicity compared with wild strain.This study provides a gene knockout technique for the study of EHEC gene function and of the relationships between RelEz5150 and EHEC pathogenicity.关键词
肠出血性大肠杆菌/Red重组系统/毒素/RelE/致病性Key words
enterohemorrhage E.coli/λRed recombinant system/toxin/RelE/pathogenicity分类
生物科学引用本文复制引用
何志利,谭琳琳,王建新,李崭,陈芳红,李涛,王慧..肠出血性大肠杆菌z5150毒素基因在细胞黏附过程中的作用[J].军事医学,2017,41(12):978-982,5.基金项目
国家自然科学基金资助项目(81401643) （81401643）
国家重点基础研究发展计划资助项目(2015CB554202) （2015CB554202）
