肠炎沙门菌YbjX基因缺失株的构建及生物学特性研究
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摘要
为研究肠炎沙门菌外膜蛋白YbjX对该菌致病作用的影响,本研究利用λ-Red同源重组系统构建了肠炎沙门菌(C50336)YbjX基因缺失突变株(C50336ΔYbjX),并对其生物学特性进行分析,探究YbjX蛋白在肠炎沙门菌致病过程中的作用。结果显示,缺失株C50336ΔYbjX与野生型菌株和回复菌株相比,其生长速度、生化特性无明显差异,但其在血清中和蛋清中的存活能力明显下降。此外,药敏试验结果表明YbjX基因缺失会引起沙门菌药物敏感性增强。进一步的,动物致病性实验表明,YbjX基因缺失会引起沙门菌毒力和巨噬细胞内存活能力的严重下降。本研究证实YbjX基因与肠炎沙门菌毒力密切相关,为进一步阐释肠炎沙门菌感染致病机制奠定基础。
To investigate the role of outer membrane protein YbjX in Salmonella enteritidis infection, we generated a YbjX-knockout mutant by λ-Red homologous recombination, and examined the growth characteristics, biochemical characteristics, antibiotic-resistance and virulence of YbjX deleted mutant. The data showed that, the YbjX-knockout didn't affect the growth characteristics and biochemical characteristics C50336 strains. But compared to the wild type and complemented strains, the YbjX-deficient mutant displayed sharply impaired survival in chicken and egg albumen. Sensitivity to Polymyxin B and some other antibiotics were significantly increased. The apparently attenuated virulence and impaired survival in macrophage were also observed in YbjX-deleted mutant. This study determined the close relationship of YbjX and Salmonella's virulence and provided insight into the underlying mechanism of Salmonella caused infection.
To investigate the role of outer membrane protein YbjX in Salmonella enteritidis infection, we generated a YbjX-knockout mutant by λ-Red homologous recombination, and examined the growth characteristics, biochemical characteristics, antibiotic-resistance and virulence of YbjX deleted mutant. The data showed that, the YbjX-knockout didn't affect the growth characteristics and biochemical characteristics C50336 strains. But compared to the wild type and complemented strains, the YbjX-deficient mutant displayed sharply impaired survival in chicken and egg albumen. Sensitivity to Polymyxin B and some other antibiotics were significantly increased. The apparently attenuated virulence and impaired survival in macrophage were also observed in YbjX-deleted mutant. This study determined the close relationship of YbjX and Salmonella's virulence and provided insight into the underlying mechanism of Salmonella caused infection.
引文
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[2] Patrick ME,Adcock PM,Gomez TM,et al.Salmonella enteri-tidis infections,United States,1985-1999[J].Emerg Infect Dis,2004,10(1):1-7.DOI:10.3201/eid1001.020572
[3] Braden CR.Salmonella enterica serotype Enteritidis and eggs:a national epidemic in the United States[J].Clin Infect Dis,2006,43(4):512-517.DOI:10.1086/505973
[4] Mckelvey JA,Yang M,Jiang Y,et al.Salmonella enterica serovar enteritidis antimicrobial peptide resistance genes aid in defense against chicken innate immunity,fecal shedding,and egg deposition[J].Infect Immun,2014,82(12):5185-5202.DOI:10.1128/IAI.02387-14
[5] 王佐强,姚玉峰.沙门菌效应蛋白对宿主细胞的影响及分子机制[J].微生物学报:1-11.
[6] Song X,Li C,Qi K,et al.The role of the outer membrane protein gene YbjX in the pathogenicity of avian pathogenic Escherichia coli[J].Avian Pathol,2018,47(3):294-299.DOI:10.1080/03079457.2018.1448053
[7] 张志强,李永慧,吴同垒,等.致泥鳅点状出血征温和气单胞菌的分类鉴定[J].中国兽医学报.2017(6):1070-1076.
[8] Wang CY,Wang SW,Huang WC,et al.Prc contributes to Escherichia coli evasion of classical complement-mediated serum killing[J].Infect Immun,2012,80(10):3399-3409.DOI:10.1128/IAI.00321-12
[9] 蒋惠源,曹鸣辉,曹学松,等.非编码小RNA(sraB)调控肠炎型沙门菌的抗蛋清抑菌作用[J].微生物学报.2010(11):1537-1544.
[10] Monack DM.Salmonella persistence and transmission strategies[J].Curr Opin Microbiol,2012,15(1):100-107.DOI:10.1016/j.mib.2011.10.013
[11] Zwir I,Latifi T,Perez JC,et al.The promoter architectural landscape of the Salmonella PhoP regulon[J].Mol Microbiol,2012,84(3):463-485.DOI:10.1111/j.1365-2958.2012.08036.x
[12] Fu J,Qi L,Hu M,et al.Salmonella proteomics under oxidative stress reveals coordinated regulation of antioxidant defense with iron metabolism and bacterial virulence[J].J Proteomics,2017,157:52-58.DOI:10.1016/j.jprot.2017.02.004
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