安徽农业大学学报2012，Vol.39Issue(6)：854-858,5.
利用Red重组系统敲除APEC毒力岛irp2基因
Deletion of irp2 gene of avian pathogenic E.coli strain with HPI by red recombination
摘要
Abstract
The aim of this research was to construct irp2 gene deletion mutant of avian pathogenic E.coli (APEC) which harbored the HPI by using Red recombination system. PCR products were obtained by using primers with 51 bp extension homologous to irp2 using template plasmid pKD3 which carrying chloramphenicol resistance gene flanked by FRT sites. With the plasmid pKD46 mediated Red recombinant system, the PCR products were transformed into APEC (CE02) by electroporation to recombine irp2 gene, and the recombinant was selected on chloramphenicol agar plate. Then the plasmid pCP20 which encoded the Flp recombinase was introduced into the recombinant to eliminate chloramphenicol resistance gene. The results showed that gene of irp2 deletion mutant of APEC was successfully constructed. The recombinant strains would be the foundation for the further research on the role of irp2 gene in pathogenic mechanism of APEC.关键词
禽致病性大肠杆菌/HPI毒力岛/Red同源重组酶/irp2/缺失Key words
avian pathogenic E.coli/ high pathogenicity island/ Red recombinase/ irp2\ deletion分类
农业科技引用本文复制引用
李叶芳,涂健,邵颖,刘红梅,祁克宗..利用Red重组系统敲除APEC毒力岛irp2基因[J].安徽农业大学学报,2012,39(6):854-858,5.基金项目
国家自然科学基金(30871851)资助. （30871851）
