Construction and functional studies of uropathogenic E. coli strains with ompT gene knockout.
- Author:
Tie ZHAO
1
;
Xingxing FANG
;
Xiaolu LIU
;
Liang PENG
;
Min LONG
;
Wenbing ZHANG
;
Jun LUO
;
Hong CAO
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Bacterial Proteins; genetics; Cell Line; Escherichia coli Infections; microbiology; Escherichia coli Proteins; genetics; Gene Knockout Techniques; Humans; Mice; Mice, Inbred C57BL; Porins; genetics; Urinary Tract Infections; microbiology; Uropathogenic Escherichia coli; genetics
- From: Journal of Southern Medical University 2012;32(7):956-959
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo explore the role of ompT gene in uropathogenic E. coli (UPEC) CFT073 strain in urinary tract infection (UTI).
METHODSAn ompT deletion mutant (COTD) was generated by λ Red recombineering in the UPEC CFT073 strain, which was characterized by PCR and sequencing. C57B/L6 mouse models of acute UTI with the mutant and wild-type strains were established to compare the colonization abilities of the two strains in the bladder. The adhesion of CFT073 mutant to human unthelial 5637 cells was also investigated in vitro.
RESULTSPCR and DNA sequencing confirmed the loss of ompT gene in the mutant COTD. The in vitro adhesion rate of the mutant strain COTD to 5637 cells was (6.7±2.2)%, significantly lower than that of (8.3±1.9)% of the wild-type strain (P<0.05). In the murine models of acute UTI, the mutant strain showed a mean colonization number of about (17±8)×10⁴ cfu, which was significantly lower than that of (7∓2)×10⁵ cfu of the wide-type CFT073 strain (P<0.05).
CONCLUSIONOmpT gene can be involved in the colonization of UPEC in the bladder tissue and plays an important role in the pathogenesis of UPEC-induced UTI.
