Construction of uropathogenic Escherichia coli strain with ppk1 gene deletion and study on its biolog-ical properties
10.3760/cma.j.issn.0254-5101.2013.07.010
- VernacularTitle:尿路致病性大肠杆菌ppk1基因缺失株的构建及其生物学特性研究
- Author:
Su LUO
;
Liang PENG
;
Jiayun PAN
;
Xiaoman WU
- Publication Type:Journal Article
- Keywords:
Urinary tract infection;
Escherichia coli;
Gene knockout;
Polyphosphate kinase 1
- From:
Chinese Journal of Microbiology and Immunology
2013;(7):531-536
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a Polyphosphate kinase 1 ( ppk1) gene deletion mutant of uro-pathogenic Escherichia coli (E.coli) CFT073, and to explore the biological properties of the mutant strain . Methods The ppk1 gene deletion strain (△pk1) was constructed based on CFT073 E.coli strain by usingλRed homologous recombination technology .A comparison analysis was conducted on adhesive and invasive abilities between CFT073 wild type strain and △pk1 strain in in vitro model of human bladder cancer epithe-lial cell 5637 .Crystal violet staining method was used to evaluate the influences of ppk1 gene deletion on biofilm formation.Results The CFT073 ppk1 deletion mutant strain was successfully constructed .Com-pared with the wild type strain ,△pk1 strain showed impaired adhesive and invasive abilities to 5637 cells. Moreover , the absorbance values of crystal violet at 570 nm at each time point of the mutant strain group were also lower than those of the wild-type strain group .Conclusion The ppk1 gene deletion mutant of uro-pathogenic E.coli CFT073 could be successfully constructed by Red homologous recombination technology and its biological properties indicates that ppk1 gene plays an important role in the pathogenesis of uropatho-genic E.coli infection through regulating the abilities of adhesion , invasion and biofilm formation .
