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副溶血弧菌Ⅲ型分泌系统vscG基因缺失株的构建及其生物学特性
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国家自然科学基金(31871893)


Construction and characterization of the vscG gene mutant strain of Vibrio parahaemolyticus type Ⅲ secretion system
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    摘要:

    【背景】副溶血弧菌(Vibrio parahaemolyticus)是一种重要的食源性病原微生物,Ⅲ型分泌系统(type Ⅲ secretion system,T3SS)在致病过程中发挥重要作用。vscG基因编码的VscG蛋白是Ⅲ型分泌系统的伴侣蛋白。vscG基因在副溶血弧菌中的生物学功能尚未确定。【目的】构建副溶血弧菌vscG基因的缺失株和回补株,研究vscG基因对副溶血弧菌生物学特性的影响。【方法】在副溶血弧菌POR-1菌株基础上利用同源重组法构建vscG基因的缺失株ΔvscG和回补株CΔvscG,分析比较POR-1、ΔvscG和CΔvscG在生长性能、生物被膜形成能力、运动性、溶血活性、细胞黏附和细胞毒性等方面存在的差异。【结果】与POR-1菌株相比,缺失株ΔvscG和回补株CΔvscG的生长特性和溶血活性无明显差异,缺失株ΔvscG的生物被膜形成能力下降,运动性较POR-1菌株和回补株CΔvscG增强。细胞感染试验显示,vscG基因的缺失明显降低了副溶血弧菌对HeLa细胞的黏附和毒性作用。【结论】vscG基因影响副溶血弧菌生物被膜的形成和运动性,在该菌黏附细胞和发挥细胞毒性过程中具有重要作用,为进一步探讨副溶血弧菌T3SS的致病机理奠定基础。

    Abstract:

    [Background] Vibrio parahaemolyticus is an important food-borne pathogenic microorganism. Type Ⅲ secretion system (T3SS) plays an important role in the pathogenic process of V. parahaemolyticus. The VscG protein encoded by the vscG gene is a chaperone protein in T3SS. The biological function of vscG gene has not been determined. [Objective] To analysis the effect of the vscG gene on the biological characteristics, a deletion strain and complement strain of the vscG gene were constructed and characterized. [Methods] We constructed the vscG gene mutant strain and complemented strain of POR-1 strain using the homologous recombination method. Then we analyzed the growth characteristics, biofilm formation, motility, hemolytic activity, cells adhesion and cytotoxicity. [Results] Compared with POR-1 strain, the growth characteristics and hemolytic activity of the deletion strain ΔvscG and the complementing strain CΔvscG were not significantly different. The biofilm formation ability of ΔvscG was reduced. The motility was enhanced of ΔvscG as compared with POR-1 and CΔvscG. Cell infection tests showed that the deletion of the vscG gene significantly reduced the adhesion and toxicity of V. parahaemolyticus to HeLa cells. [Conclusion] The vscG gene affects the motility and formation of the biofilm of V. parahaemolyticus, it plays an important role in the adhesion and cytotoxicity to HeLa cells, laying a foundation for further exploring the pathogenic mechanism of T3SS in V. parahaemolyticus.

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薛娇,廉乐乐,李婉君,任建鸾,汤芳,薛峰,戴建君. 副溶血弧菌Ⅲ型分泌系统vscG基因缺失株的构建及其生物学特性[J]. 微生物学通报, 2021, 48(6): 2101-2110

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  • 在线发布日期: 2021-06-09
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