狐源沙门菌cpxR基因缺失菌株的构建及部分表型研究
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摘要
为研究狐源沙门菌双组份系统CpxR/A在细菌耐药和抗血清杀伤中的作用,本研究利用λ-Red重组技术构建了cpxR基因缺失株S-ΔcpxR,评价其在生物被膜形成、耐药、抗血清杀伤和细菌毒力中的重要作用。结果显示,与野生型相比,cpxR基因缺失株的生长速度、生化特性没有改变,但是生物被膜形成能力显著降低,对阿莫西林、阿米卡星和恩诺沙星的敏感性增加,血清中存活率和对小鼠的毒力显著降低。结果表明,沙门菌CpxR与毒力相关,为狐源沙门菌基因缺失苗的研究奠定基础。
To investigate the roles of two-component regulatory systems CpxR/A in the drug resistance and serum-killing of Salmonella isolated from fox,the related experiments was performed,after the knockout of cpxR gene by λ-Red recombination.The data demonstrated that,the cpxR-deficient mutant showed no difference in growth characteristics and biochemical characteristics,compared with wild type strain.However,the mutant strain showed increased sensibility to antibiotics including amoxicillin,amikacin and enrofloxacin,survival defection in serum and decrease bacterial virulence to mice.This study suggested that CpxR contributed to the virulence and will provide the foundation for the vaccine of Salmonella.
To investigate the roles of two-component regulatory systems CpxR/A in the drug resistance and serum-killing of Salmonella isolated from fox,the related experiments was performed,after the knockout of cpxR gene by λ-Red recombination.The data demonstrated that,the cpxR-deficient mutant showed no difference in growth characteristics and biochemical characteristics,compared with wild type strain.However,the mutant strain showed increased sensibility to antibiotics including amoxicillin,amikacin and enrofloxacin,survival defection in serum and decrease bacterial virulence to mice.This study suggested that CpxR contributed to the virulence and will provide the foundation for the vaccine of Salmonella.
引文
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[11] LIMA B P,LENNON C W,ROSS W,et al.In vitro evidence that RNA Polymerase acetylation and acetyl phosphate-dependent CpxR phosphorylation affect cpxP transcription regulation[J].FEMS Microbiol Lett,2016,363(5):w11.
[12] GANGAIAH D,RATERMAN E L,WU H,et al.Both MisR (CpxR) and MisS (CpxA) are required for neisseria gonorrhoeae infection in a murine model of lower genital tract infection[J].Infect Immun,2017,85(9):e00307-17.
[13] CERMINATI S,GIRI G F,MENDOZA J I,et al.The CpxR/CpxA system contributes to Salmonella gold-resistance by controlling the GolS-dependent gesABC transcription[J].Environ Microbiol,2017,19(10):4035-4044.
[2] 徐乐,周晓辉,何晓亮.革兰阴性菌的Cpx双组分调控系统[J].微生物学报.2014(3):269-275.
[3] 吴同垒,张志强,贾清辉,等.白痢沙门菌CpxR的生物信息学分析和原核表达[J].中国兽医学报,2017,37(10):1891-1895.
[4] HUJA S,OREN Y,BIRAN D,et al.Fur is the master regulator of the extraintestinal pathogenic Escherichia coli response to serum[J].MBio,2014,5(4):e01460-14.
[5] KURABAYASHI K,HIRAKAWA Y,TANIMOTO K,et al.Role of the CpxAR two-component signal transduction system in control of fosfomycin resistance and carbon substrate uptake[J].J Bacteriol,2014,196(2):248-256.
[6] DE LA CRUZ M A,PEREZ-MORALES D,PALACIOS I J,et al.The two-component system CpxR/A represses the expression of Salmonella virulence genes by affecting the stability of the transcriptional regulator HilD[J].Front Microbiol,2015(6):807.
[7] TIAN Z X,YI X X,CHO A,et al.CpxR activates MexAB-OprM efflux pump expression and enhances antibiotic resistance in both laboratory and clinical nalB-Type isolates of pseudomonas aeruginosa[J].PLoS Pathog,2016,12(10):e1005932.
[8] HOLDEN K M,BROWNING G F,NOORMOHAMMADI A H,et al.TonB is essential for virulence in avian pathogenic Escherichia coli[J].Comp Immunol Microbiol Infect Dis,2012,35(2):129-138.
[9] SAMANT S,LEE H,GHASSEMI M,et al.Nucleotide biosynthesis is critical for growth of bacteria in human blood[J].PLoS Pathog,2008,4(2):e37.
[10] SERRUTO D,RAPPUOLI R,SCARSELLI M,et al.Molecular mechanisms of complement evasion:learning from staphylococci and meningococci[J].Nat Rev Microbiol,2010,8(6):393-399.
[11] LIMA B P,LENNON C W,ROSS W,et al.In vitro evidence that RNA Polymerase acetylation and acetyl phosphate-dependent CpxR phosphorylation affect cpxP transcription regulation[J].FEMS Microbiol Lett,2016,363(5):w11.
[12] GANGAIAH D,RATERMAN E L,WU H,et al.Both MisR (CpxR) and MisS (CpxA) are required for neisseria gonorrhoeae infection in a murine model of lower genital tract infection[J].Infect Immun,2017,85(9):e00307-17.
[13] CERMINATI S,GIRI G F,MENDOZA J I,et al.The CpxR/CpxA system contributes to Salmonella gold-resistance by controlling the GolS-dependent gesABC transcription[J].Environ Microbiol,2017,19(10):4035-4044.