[1]廉乐乐,薛娇,李婉君,等.副溶血弧菌Ⅲ型分泌系统1内杆状蛋白基因vscI的功能分析[J].南京农业大学学报,2021,44(3):506-513.[doi:10.7685/jnau.202003056]
 LIAN Lele,XUE Jiao,LI Wanjun,et al.Functional analysis of the inner rod protein gene vscI in Vibrio parahaemolyticus Type Ⅲ secretion system 1[J].Journal of Nanjing Agricultural University,2021,44(3):506-513.[doi:10.7685/jnau.202003056]
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副溶血弧菌Ⅲ型分泌系统1内杆状蛋白基因vscI的功能分析()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
44卷
期数:
2021年3期
页码:
506-513
栏目:
动物科学
出版日期:
2021-05-10

文章信息/Info

Title:
Functional analysis of the inner rod protein gene vscI in Vibrio parahaemolyticus Type Ⅲ secretion system 1
作者:
廉乐乐1 薛娇1 李婉君1 任建鸾1 诸葛祥凯1 汤芳1 薛峰1 戴建君12
1. 南京农业大学教育部动物健康与食品安全国际合作联合实验室/动物医学院, 江苏 南京 210095;
2. 中国药科大学, 江苏 南京 211199
Author(s):
LIAN Lele1 XUE Jiao1 LI Wanjun1 REN Jianluan1 ZHUGE Xiangkai1 TANG Fang1 XUE Feng1 DAI Jianjun12
1. Joint Laboratory of Animal Health and Food Safety International Cooperation, Ministry of Educations/College of Veterianry Medicine, Nanjing Agricultural University, Nanjing 210095, China;
2. China Pharmaceutical University, Nanjing 211199, China
关键词:
副溶血弧菌T3SS1内杆状蛋白vscI
Keywords:
Vibrio parahemolyticusT3SS1inner rod proteinvscI
分类号:
S855.1
DOI:
10.7685/jnau.202003056
摘要:
[目的] Ⅲ型分泌系统(Type Ⅲ secretion systems,T3SS)在副溶血弧菌(Vibrio parahemolyticus)致病过程中发挥重要作用。本试验旨在探讨T3SS1内杆状蛋白基因vscI的功能。[方法] 以副溶血弧菌POR-1菌株为研究对象,构建vscI缺失株ΔvscI与回补株CΔvscI,检测野生株、缺失株与回补株的细胞毒性、细胞黏附等生物学特性,分析vscI缺失对副溶血弧菌感染细胞能力的影响;通过Western blot与ELISA检测HeLa细胞内效应蛋白的易位量。[结果] 与POR-1菌株相比,缺失菌株ΔvscI在生长速度上并无显著差异,但对HeLa细胞的黏附能力下降85%。细胞毒性检测结果显示:与POR-1菌株相比,缺失株ΔvscI感染细胞组释放LDH量显著下降60%。同时与野生株POR-1感染HeLa细胞组相比,ΔvscI感染HeLa细胞组发生凋亡的细胞数量显著下降了85%。腺苷酸环化酶报告系统研究结果显示:与POR-1菌株感染组相比,ΔvscI菌株感染HeLa细胞内的cAMP含量显著下降,CyaA融合蛋白的易位量显著下降,而在回补株CΔvscI中上述生物性状均恢复。[结论] vscI提高了副溶血弧菌对细胞的黏附能力和毒力,促进细胞凋亡,同时对于T3SS1效应子的易位是必需的。
Abstract:
[Objectives] The structural components and the biological functions of Vibrio parahemolyticus T3SS1 have not been fully described. In this study we investigated the role of gene vscI. [Methods] To do some research about the vscI,we constructed the deletion strain ΔvscI and complementation strain CΔvscI to analyze the cytotoxicity,adhesion and other biological characteristics of wild strain(POR-1),ΔvscI and CΔvscI when infected HeLa. Further,Western blot analysis and ELISA kits were used to research the translocation of effector protein in Hela cell. [Results] Compared with POR-1 infection group,the results showed that the number of apoptosis decreased significantly by about 80%(P<0.01),the adhesion decreased significantly by 85%(P<0.01),and the cytotoxicity decreased significantly by 60%(P<0.01) in ΔvscI infection group. In addition,Western blot and ELISA data indicated that the intracellular cAMP levels of the ΔvscI group was significantly lower than those of the POR-1,CΔvscI. [Conclusions] vscI improves the adhesion ability of V.parahaemolyticus to cells,and produces cytotoxicity,promotes cell apoptosis,and it is necessary for the translocation of T3SS1 effector.

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备注/Memo

备注/Memo:
收稿日期:2020-03-27。
基金项目:国家自然科学基金项目(31871893)
作者简介:廉乐乐,博士研究生。
通信作者:薛峰,教授,主要从事人畜共患病的病原学研究,E-mail:xuefeng@njau.edu.cn。
更新日期/Last Update: 1900-01-01