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[猪链球菌2型分选酶BCD基因敲除突变体的构建及体外测定]

[Construction and in vitro assay of the sortase BCD geneknock-out mutant of Streptococcus suis 2].

作者信息

Chen Hongna, Liao Hui, Wang Changjun, Pan Xiuzhen, Tang Jiaqi

机构信息

Institute of Military Medical Sciences, Nanjing Command, Nanjing 210002, China.

出版信息

Wei Sheng Wu Xue Bao. 2011 Mar;51(3):386-92.

Abstract

OBJECTIVE

Streptococcus suis 2 is an emerging zoonotic pathogen responsible for a wide range of life-threatening diseases in pigs and humans. In this study, we investigated the functionality of one of Streptococcus suis 2 sortases, known as the srtBCD.

METHODS

To obtain the isogenic mutant srtBCD, the competent cells of 05ZYH33 were subjected to electrotrans formation with recombinant plasmid based on the principle of homologous recombination. The resulting mutant strains was further confirmed by a series of PCR and reverse transcription PCR. To better assess the role of srtBCD gene in the virulence of 05ZYH33, cell adherence assays and experimental infection of mice was adopted.

RESULTS

A SrtBCD defective mutant of 05ZYH33 was found to be associated with growth curve upon cultivation in standard laboratory used in our in vitro assays. Furthermore, abolishment of the expression of srtBCD result in impaired interactions of S. suis with human laryngeal epithelial cell line. However, there is no differences when infection mice by the WT and mutant strain.

CONCLUSION

These results suggest that srtBCD are critical for the pathogen-host interaction of S. suis 2, but abolishment of srtBCD does not impair the full virulence of 05ZYH33. It is to expect that future study carried out with S. suis 2 to verification the conclusions.

摘要

目的

猪链球菌2型是一种新出现的人畜共患病原体,可导致猪和人类多种危及生命的疾病。在本研究中,我们调查了猪链球菌2型分选酶之一(称为srtBCD)的功能。

方法

为获得srtBCD基因缺失突变体,根据同源重组原理,用重组质粒对05ZYH33感受态细胞进行电转化。通过一系列PCR和逆转录PCR进一步确认所得突变株。为更好地评估srtBCD基因在05ZYH33毒力中的作用,采用细胞黏附试验和小鼠实验感染。

结果

在我们体外试验所用的标准实验室培养条件下,发现05ZYH33的SrtBCD缺陷突变体与生长曲线有关。此外,srtBCD表达的缺失导致猪链球菌与人喉上皮细胞系的相互作用受损。然而,野生型菌株和突变株感染小鼠时没有差异。

结论

这些结果表明,srtBCD对猪链球菌2型的病原体-宿主相互作用至关重要,但srtBCD的缺失并不损害05ZYH33的全部毒力。期望未来对猪链球菌2型进行研究以验证这些结论。

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