副猪嗜血杆菌<em>ptsG</em>基因缺失株的构建及其部分生物学特性分析

doi:10.3969/j.issn.1674-7968.2021.12.012

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摘要葡萄糖磷酸转移酶系统(glucose-specific phosphotransferase system, PTS^Glc)中葡萄糖特异酶ⅡBC(glucose-specific enzyme ⅡBC, EⅡBC^Glc)对细菌的代谢有着全面的影响,但EⅡBC^Glc在副猪嗜血杆菌(Glaesserella parasuis)中的作用还不清楚。本研究采用自然转化法构建副猪嗜血杆菌EⅡBC^Glc蛋白的编码基因(phosphotransferase system, ptsG)缺失株及互补株,比较野毒株和缺失株在胰蛋白胨大豆肉汤+葡萄糖培养基的生长状况、细菌形态、生物被膜形成能力、对氧化压力的耐受能力及对小鼠(Mus musculus)的毒力作用。结果表明,ptsG基因敲除后,副猪嗜血杆菌可以摄取更多的还原糖供自身生长。通过透射电镜观察,发现ΔptsG与野生株相比,菌体内部有更多的高电子密度区。此外,ptsG基因的缺失导致副猪嗜血杆菌生物被膜形成能力减弱以及对过氧化氢的耐受能力减弱。而ΔptsG与野生株对小鼠的毒力作用没有明显差异。本研究表明ptsG基因对副猪嗜血杆菌生物被膜的形成和氧化应激耐受能力具有重要的作用,为研究副猪嗜血杆菌的致病机制提供了基础资料。

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	闫雪锋
	肖五淀
	古从伟
	何曼莉
	赵明德
	何绿琴

关键词 ：副猪嗜血杆菌, PTS^Glc系统, ptsG, 生物被膜, 氧化应激

Abstract：In the glucose-specific phosphotransferase (PTS^Glc) system, glucose-specific enzyme ⅡBC (EⅡBC^Glc) has a comprehensive effect on the metabolism of bacteria, but of which the function is still unclear in Glaesserella parasuis. In this study, EⅡBC^Glc protein coding gene (ptsG) mutant and comlementary strain were constructed by natural transformation method. The growth status of wild strain SC1401 and ptsG mutant strains in the trypton soy broth (TSB)+glucose medium were compared in this study. Moreover, the bacterial morphology, biofilm formation, tolerance to oxygen pressure and virulence to mice (Mus musculus) of the SC1401 and ptsG mutant strain were measured. The results showed that the ptsG mutant strain could take in more reducing sugars for its own growth. Through transmission electron microscopy that compared with the wild strain, ptsG mutant strain had more high electron density areas inside the bacteria. In addition, the formation of biofilm and the tolerance to hydrogen peroxide were decreased in ptsG mutant strain. But there was no significant difference between SC1401 and ptsG mutant strains on mice. This study found that the ptsG gene played an important role in the formation of biofilm and oxidative stress tolerance, which will lay a foundation for the study of pathogenic mechanism in Glaesserella parasuis.

Key words： Glaesserella parasuis PTS^Glc system ptsG Biofilm Oxidative stress

收稿日期: 2021-03-26

ZTFLH:

S852.61

基金资助:泸州市人民政府与西南医科大学科技战略合作计划(2019LZXNYDJ20); 西南医科大学校级项目(2019ZQN141; 2020ZRQNA010)

通讯作者: *helvqin617@163.com

引用本文:

闫雪锋, 肖五淀, 古从伟, 何曼莉, 赵明德, 何绿琴. 副猪嗜血杆菌ptsG基因缺失株的构建及其部分生物学特性分析[J]. 农业生物技术学报, 2021, 29(12): 2387-2395.
YAN Xue-Feng, XIAO Wu-Dian, GU Cong-Wei, HE Man-Li, ZHAO Ming-De, HE Lyu-Qin. Construction of Glaesserella parasuis ptsG Mutant Strain and Analysis of Its Partial Biological Characteristics. 农业生物技术学报, 2021, 29(12): 2387-2395.

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http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2021.12.012 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2021/V29/I12/2387

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