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[假定调控蛋白STM14_3514减少鼠伤寒沙门氏菌对上皮细胞的侵袭]

[Putative regulatory protein STM14_3514 decreases Salmonella Typhimurium invasion of epithelial cells].

作者信息

Jiang Lingyan, Zhou Qixing, Wang Peisheng, Jiang Xiaohan, Feng Lu

出版信息

Wei Sheng Wu Xue Bao. 2017 Apr 4;57(4):500-12.

Abstract

OBJECTIVE

To study the function and mechanism of STM14_3514 gene that encoded in Salmonella pathogenicity island (SPI)-1 of Salmonella enterica serovar Typhimurium strain ATCC 14028.

METHODS

We constructed STM14_3514 mutant strain and a complemented strain of the mutant. Through mice experiment, attachment assays, invasion assays, macrophage replication assays, western blot, and Quantitative real-time PCR analysis (qRT-PCR), we compared the virulence of the mutant strain to that of the wild-type 14028.

RESULTS

STM14_3514 mutant shows increased virulence to mice, and the bacterial number of STM14_3514 mutant in liver, spleen, and ileum was more abundant than that of the wild-type strain. The increased virulence of STM14_3514 mutant is caused by its elevated invasion ability to epithelial cells (>2-fold and P<0.05). qRT-PCR and western blot results show that STM14_3514 reduced the expression of HilA and another SPI-1invasion locus. Moreover, the repression of HilA by STM14_3514 is mediated by HilC.

CONCLUSION

STM14_3514 is a negative regulator in SPI-1, which can repress HilA and SPI-1invasion locus through HilC, and possibly contribute to the repression on SPI-1 after bacterial invasion.

摘要

目的

研究鼠伤寒沙门氏菌菌株ATCC 14028的沙门氏菌致病岛(SPI)-1中编码的STM14_3514基因的功能及机制。

方法

构建STM14_3514突变株及其互补菌株。通过小鼠实验、黏附试验、侵袭试验、巨噬细胞复制试验、蛋白质印迹法和定量实时PCR分析(qRT-PCR),比较突变株与野生型14028的毒力。

结果

STM14_3514突变株对小鼠的毒力增强,其在肝脏、脾脏和回肠中的细菌数量比野生型菌株更多。STM14_3514突变株毒力增强是由于其对上皮细胞的侵袭能力提高(>2倍且P<0.05)。qRT-PCR和蛋白质印迹结果表明,STM14_3514降低了HilA和另一个SPI-1侵袭位点的表达。此外,STM14_3514对HilA的抑制作用是由HilC介导的。

结论

STM14_3514是SPI-1中的负调节因子,可通过HilC抑制HilA和SPI-1侵袭位点,可能在细菌侵袭后对SPI-1起抑制作用。

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