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A Novel Phage Element of Salmonella enterica Serovar Enteritidis P125109 Contributes to Accelerated Type III Secretion System 2-Dependent Early Inflammation Kinetics in a Mouse Colitis Model

机译:肠炎沙门氏菌肠炎沙门氏菌P125109的新型噬菌体元素有助于加速III型分泌系统2依赖的小鼠结肠炎模型中的早期炎症动力学。

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摘要

Salmonella enterica subsp. I serovar Enteritidis exhibits type III secretion system 2 (TTSS2)-dependent early colonization and inflammation kinetics faster than those of closely related S. enterica serovar Typhimurium. To investigate the accelerated TTSS-2-dependent pathogenic potential of S. Enteritidis, we focused on its genome. Results of a previously published comparative genomic study revealed the presence of mutually exclusive genes in both serovars. In this study, we investigated the roles of six S. Enteritidis-specific genes in vivo by using differential fluorescence induction (DFI) through putative gene-specific promoters. The promoter construct associated with the gene locus SEN1140 induced green fluorescent protein (GFP) expression in the gut lumen, lamina propria, mesenteric lymph nodes, and related systemic organs. To further investigate the potential role of SEN1140, we compared a SEN1140 deletion mutant with S. Typhimurium in a TTSS1-deficient background. Interestingly, the S. Enteritidis mutant lacking SEN1140 did not show the unique TTSS-2-dependent early colonization and inflammation kinetic phenotype of S. Typhimurium. Consistent with this result, complementation of SEN1140 restored the TTSS-2-dependent accelerated inflammatory potential of S. Enteritidis. This report presents a suitable screening strategy that uses a combination of DFI, fluorescence-activated cell sorting, quantitative PCR, and wild-type isogenic tagged-strain techniques to explore the unique roles of S. Enteritidis-specific genes in bacterial pathogenesis.
机译:肠沙门氏菌亚种。肠炎沙门氏菌表现出III型分泌系统2(TTSS2)依赖的早期定植和炎症动力学比密切相关的肠炎沙门氏菌鼠伤寒沙门氏菌。为了研究肠炎沙门氏菌的TTSS-2依赖性致病潜力,我们将重点放在其基因组上。先前发表的比较基因组研究的结果表明,两种血清型中都存在互斥基因。在这项研究中,我们通过使用推定的基因特异性启动子的差分荧光诱导(DFI),研究了六个肠炎沙门氏菌特异性基因在体内的作用。与基因位点SEN1140相关的启动子构建体诱导了肠腔,固有层,肠系膜淋巴结和相关全身器官中的绿色荧光蛋白(GFP)表达。为了进一步研究SEN1140的潜在作用,我们在TTSS1缺失的背景下比较了SEN1140缺失突变体和鼠伤寒沙门氏菌。有趣的是,缺乏SEN1140的肠炎沙门氏菌突变体未显示鼠伤寒沙门氏菌独特的TTSS-2依赖性早期定植和炎症动力学表型。与该结果一致,SEN1140的互补恢复了肠炎沙门氏菌的TTSS-2依赖性加速的炎症潜能。本报告提出了一种合适的筛选策略,该方法结合使用DFI,荧光激活细胞分选,定量PCR和野生型等基因标记菌株技术,以探索肠炎沙门氏菌特异性基因在细菌发病中的独特作用。

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