目的:研究伤寒沙门菌ssrB基因在氧应激早期对其他基因表达的调节。方法:通过同源重组的方法利用自杀质粒制备伤寒沙门菌ssrB基因缺陷变异株;采用伤寒沙门菌全基因组芯片比较野生株和ssrB基因缺陷变异株在氧应激早期的基因表达差异,并对其中部分表达差异基因进行实时定量PCR验证;利用HeLa细胞进行细菌侵袭实验,研究ssrB基因对伤寒沙门菌侵袭能力的影响。结果:成功制备伤寒沙门菌ssrB基因缺陷变异株;基因芯片结果分析显示,在氧应激早期,与野生株相比,伤寒沙门菌ssrB基因缺陷变异株有68个基因表达下调,有20个基因表达上调,其中与侵袭相关的基因表达下调。实时定量PCR与芯片结果一致。ssrB基因缺陷变异株侵袭上皮细胞的能力仅为野生株的34.6%。结论:SsrB在伤寒沙门菌氧应激早期对基因表达起重要调节作用,并且能够增强伤寒沙门菌侵袭上皮细胞的能力。
Objective: To investigate the regulation of ssrB on gene expression of Salmonella enterica serovar Typhi(S.Typhi) at early-stage of oxidative stress.Methods: The ssrB deleted mutant of S.Typhi was generated through homologous recombination mediated by suicide plasmid;the gene expression profiles of the wild-type strain and the ssrB deleted mutant at early-stage of oxidative stress was investigated by genomic microarray assay;qRT-PCR was performed to further validate the results of microarray assay.The HeLa cells were invaded by S.Typhi to explore the influence of ssrB on invasion of epithelial cells.Results: The ssrB deleted mutant of S.Typhi was prepared successfully;analysis of genomic assay showed that,compared to the wild-type strain,68 genes were up-regulated and 20 genes were down-regulated in the ssrB deleted mutant at early stage of oxidative stress.The results of qRT-PCR assay were consistent with the results of microarray assay.The ability of the ssrB deleted mutant to invade the epithelial cells was only 34.6% of the wild-type strain.Conclusion: SsrB of S.Typhi played an important role in regulating gene expression at early stage of oxidative stress and enhanced the ability of S.Typhi to invade epithelial cells.