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微生物学通报

副溶血弧菌染色体上毒素-抗毒素系统YefM-YoeB的鉴定
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国家自然科学基金(31802210);中国博士后科学基金(2018M630615);江苏省高校自然科学基金(18KJB230007)


Identification of the chromosomal YefM-YoeB toxin-antitoxin system in Vibrio parahemolyticus
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    摘要:

    【背景】副溶血弧菌是一种重要的食源性病原菌,给公众健康带来严重危害。毒素-抗毒素系统广泛存在于细菌和古生菌基因组中,具有重要的生物学功能。【目的】在副溶血弧菌中鉴定新的毒素-抗毒素系统,为从毒素-抗毒素系统角度探讨该菌致病性和耐药性的分子机制奠定基础。【方法】通过在线工具预测副溶血弧菌染色体上的假定II型毒素-抗毒素系统;通过生长曲线分析和稀释点板实验检测假定毒素对大肠杆菌的毒性作用及相应抗毒素的抗毒性作用;通过反转录PCR确定毒素和抗毒素基因是否共转录;通过生物信息学分析确定新鉴定毒素-抗毒素系统的同源蛋白;通过LacZ报告实验确定抗毒素及毒素-抗毒素复合物对自身启动子的调控作用。【结果】副溶血弧菌染色体中编码6个假定II型毒素-抗毒素系统;基因vp1820的表达产物(VP1820)对大肠杆菌具有杀菌活性,vp1821的表达产物(VP1821)能中和VP1820的毒性;基因vp1821和vp1820共转录;vp1821-vp1820编码YefM-YoeB毒素-抗毒素系统;抗毒素YefM正调控启动子,YefM-YoeB复合物负调控启动子。【结论】在副溶血弧菌中鉴定了一个新的II型毒素-抗毒素系统,即YefM-YoeB,为进一步研究该系统对副溶血弧菌致病性和耐药性的影响奠定了基础。

    Abstract:

    [Background] Vibrio parahemolyticus is an important food-borne pathogen causing severe threats to public health. Toxin-antitoxin (TA) systems are widely distributed in the genomes of bacteria and archaea, and possess important biological functions. [Objective] To identify novel TA systems in V. parahemolyticus, and lay the foundation for exploring the underlying mechanism of bacterial pathogenicity and drug resistance from the aspect of TA systems. [Methods] The putative chromosomal type II TA systems in V. parahemolyticus were predicted by a web-based tool. The toxic effects of the putative toxins on Escherichia coli and the antitoxic effects of the corresponding antitoxins were assessed by growth curves analyses and spot dilution assays. Reverse transcription PCR was used to determine whether the genes encoding the toxin and antitoxin were co-transcribed. The homologous proteins of the newly identified TA system were determined by bioinformatics analysis. The regulation of their own promoters by the antitoxin and antitoxin-toxin complex was detected by LacZ reporter assay. [Results] Six putative chromosomal TA systems were identified in V. parahemolyticus. The product of the vp1820 gene (VP1820) had bactericidal activity against E. coli, which could be counteracted by the product of the vp1821 gene (VP1821). The genes vp1821 and vp1820 are co-transcribed. The vp1821-vp1820 locus encodes the YefM-YoeB TA system. The YefM antitoxin positively regulates the promoter, while the YefM-YoeB complex negatively regulates the promoter. [Conclusion] This study identifies a novel type II TA system in V. parahemolyticus, namely YefM-YoeB, and lays the foundation for further research on the role of this system in the pathogenicity and drug resistance of V. parahemolyticus.

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卫慢,邱军,贾梦蝶,郑成坤. 副溶血弧菌染色体上毒素-抗毒素系统YefM-YoeB的鉴定[J]. 微生物学通报, 2021, 48(6): 2061-2072

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  • 在线发布日期: 2021-06-09
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