南方医科大学学报Issue(6)：904-908,5.DOI:10.3969/j.issn.1673-4254.2014.06.30
肠出血型大肠埃希菌O157：H7 ppk基因缺失株的构建及其生物学特性分析
Construction and characterization of enterohemorrhagic Escherichia coli O157:H7 ppk-deleted strain
摘要
Abstract
Objective To construct enterohemorrhagic Escherichia coli (EHEC) O157: H7 ppk gene deletion strains and study its biological characteristics. Methods The gene fragment of kanamycin resistance was amplified using a pair of homologous arm primers whose 5' and 3' ends were homologous with ppk gene and kanamycin resistance gene, respectively. EHEC O157:H7 EDL933w competent strains were prepared and transformed via electroporation with the amplification products. The ppk gene was replaced by kanamycin resistance gene using pKD46-mediated Red recombination system. The recombinant strain was confirmed by PCR and sequencing, and its morphology, growth ability and adhesion were assessed using Gram staining, OD60 value and Giemsa staining. Results and Conclusion We established a ppk-deleted EHEC O157:H7 EDL933w strain with kanamycin resistance and compared the biological characteristics of the wild-type and mutant strains, which may facilitate further study of the regulatory mechanism of ppk gene.关键词
EHEC O157:H7/ppk基因/Red重组系统/基因突变Key words
enterohemorrhagic Escherichia coli O157:H7/ppk gene/Red recombination system/gene mutation引用本文复制引用
韩朋,孙琦,赵素慧,张其威,万成松..肠出血型大肠埃希菌O157：H7 ppk基因缺失株的构建及其生物学特性分析[J].南方医科大学学报,2014,(6):904-908,5.基金项目
国家自然科学基金（81371765）；广东省自然科学基金（S2013010014393）@@@@Supported by National Natural Science Foundation of China (81371765) （81371765）
