鼠伤寒沙门菌cpxR和acrB双基因缺失菌株的构建及其对抗菌药物敏感性分析

doi:10.11843/j.issn.0366-6964.2016.03.024

Abstract

Abstract:

The aim of this study was to explore the role of cpxR gene in the multidrug resistance of Salmonella enterica Serovar Typhimurium and exclude a possibility that acrB masks the effect of cpxR on the other efflux pumps or outer membrane proteins.A acrB gene deletion strain JSΔacrB was constructed by the method of one-step inactivation of chromosomal genes on basis of Red homologous recombination system.A double gene deletion strain JSΔacrBΔcpxR∷kan was constructed by phage P22-mediated transduction with strain JSΔcpxR∷kan constructed in the previous study as the donor and strain JSΔacrB as the recipient.Meanwhile，the cpxR complemental strain JSΔacrBΔcpxR∷kan/pcpxR was prepared through introducting the expression plasmid pBAD-CpxR into the double gene mutant.At last the minimal inhibitory concentrations(MICs) of 10 kinds of commonly used representative antibacterial agents for JS and the above strains constructed in this study were determined with broth microdilution method.The double gene deletion strain and related cpxR complemental strain were constructed successfully.Strain JSΔacrB showed 32-，4-，4-，16-，32-，4-，2- and 128-fold decrease in the MICs of doxycycline，gentamycin，amikacin，ciprofloxacin，enrofloxacin，florfenicol，ceftriaxone and ceftiofur respectively，compared to the parent strain JS.Strain JSΔacrBΔcpxR∷kan showed 2-，8-，2- and 2-fold decrease in the MICs of doxycycline，gentamicin，amikacin，and ceftiofur compared to strain JSΔacrB.While the complemental strain JSΔacrBΔcpxR∷kan/pcpxR showed 4-，16-，2- and 4-fold increase in the MICs of the above four kinds of antibacterial agents compared to strain JSΔacrBΔcpxR∷kan.In conclusion，cpxR is able to regulate the antimicrobial susceptibility of S.enterica Serovar Typhimurium to doxycycline，gentamicin，amikacin，and ceftiofur.

CLC Number:

S852.612

HUANG Hui，LIU Bao-guang，SUN Ya-wei，MA Cai-hui，CHEN Li-peng，YUAN Li，PAN Yu-shan，HU Gong-zheng. Construction of cpxR and acrB Double Gene Deletion Strain of Salmonella enterica Serovar Typhimurium and Analysis of Its Susceptibility to Antibacterial Agents[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(3): 595-602.

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Construction of cpxR and acrB Double Gene Deletion Strain of Salmonella enterica Serovar Typhimurium and Analysis of Its Susceptibility to Antibacterial Agents

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