Construction of blaNDM-1gene deletion mutant of Enterobacter cloacae and analysis of its biological characteristics
10.3760/cma.j.issn.0254-5101.2017.12.003
- VernacularTitle:阴沟肠杆菌blaNDM-1基因敲除株的构建及其生物学特性
- Author:
Jing YAO
1
;
Na DU
;
Shumin LIU
;
Min NIU
;
Mengshuang ZHANG
;
Ciyan CHEN
;
Yan DU
Author Information
1. 650032,昆明医科大学第一附属医院医学检验科 云南省实验诊断研究所 云南省检验医学重点实验室
- Keywords:
Enterobacter cloacae;
blaNDM-1gene;
Red homologous recombination;
Gene knockout
- From:
Chinese Journal of Microbiology and Immunology
2017;37(12):897-902
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a blaNDM-1gene deletion mutant of Enterobacter cloacae and to analyze its biological characteristics. Methods The blaNDM-1gene deletion mutant was constructed by using Red homologous recombination technology and verified by PCR and RT-qPCR. Antimicrobial susceptibility profiles,growth curves and in vitro competition abilities of the original strain and the blaNDM-1gene deletion mutant were analyzed. Results PCR,DNA sequencing and RT-qPCR showed that the blaNDM-1gene dele-tion mutant was successfully constructed. Antimicrobial susceptibility test showed that the original strain was resistant to imipenem,meropenem and ertapenem, while the blaNDM-1gene deletion mutant was sensitive to all. The original strain and the blaNDM-1gene deletion mutant had similar growth curves in Luria-Bertani liq-uid medium. In vitro competition experiment revealed that the competitive index of them was 0.69. Conclu-sion Red homologous recombination technology can be used to knockout the blaNDM-1gene of Enterobacter cloacae,which is associated with antimicrobial resistance and competitiveness.
