Construction of a Vibrio anguillarum flagellin B mutant and analysis of its immuno-stimulation effects on Macrobrachium rosenbergii
文献类型： 外文期刊
作者： Gao, Quanxin 1 ; Yi, Shaokui 1 ; Luo, Jinping 1 ; Xing, Qianqian 1 ; Lv, Jiali 1 ; Wang, Panhuang 1 ; Wang, Cuihua 2 ; Li 1 ;
作者机构： 1.Huzhou Univ, Coll Life Sci, Huzhou Cent Hosp,Zhejiang Prov Key Lab Aquat Reso, Key Lab Aquat Anima Genet Breeding & Nutr,Chinese, Huzhou 313000, Peoples R China
2.Chinese Acad Fishery Sci, East China Sea Fisheries Res Inst, Key Lab Marine & Estuarine Fisheries, Minist Agr, Shanghai 200090, Peoples R China
关键词： Vibrio anguillarum; Flagellin B; Toxicity
期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES （ 影响因子：6.953； 五年影响因子：6.737 ）
ISSN： 0141-8130
年卷期： 2021 年 174 卷
页码：
收录情况： SCI
摘要： Vibrio anguillarum is a globally distributed aquatic pathogen, and its flagellin B (FlaB) protein can evoke innate immune responses in hosts. In order to explore the role of FlaB in V. anguillarum infection, we constructed a FlaB-deficient mutant using overlapping PCR and two-step homologous recombination, and gene sequencing confirmed successful knockout of the FlaB gene. Scanning electron microscopy showed no significant differences in the morphological structure of the flagellum between wild-type and FlaB-deficient strains. The mutant was subsequently injected into the freshwater prawn (Macrobrachium rosenbergii) to explore its pathogenicity in the host, and expression of myeloid differentiation factor 88, prophenoloxidase, catalase, superoxide dismutase and glutathione peroxidase was investigated by real-time PCR. The results showed that deletion of FlaB had little effect on V. anguillarum-induced expression of these immune-related genes (p > 0.05). In general, the FlaB mutant displayed similar flagella morphology and immune characteristics to the wild-type strain, hence we speculated that knockout of FlaB might promote the expression and function of other flagellin proteins. Furthermore, this study provides a rapid and simple method for obtaining stable mutants of V. anguillarum free from foreign plasmid DNA. (C) 2021 Elsevier B.V. All rights reserved.
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