王俊霖,招茵,苏茵茵,周诗慧,曾福源,谢妙,王娜,简纪常,庞欢瑛

• 1:广东海洋大学水产学院//广东省水产经济动物病原生物学及流行病学重点实验室//广东省教育厅水产经济动物病害控制重点实验室
• 2:中国检验检疫科学研究院

摘要(Abstract)：

【目的】构建溶藻弧菌(Vibrio alginolyticus)Ⅲ型分泌系统(Type Ⅲ secretion system,T3SS)中的调控蛋白操纵子ExsD基因缺失株,研究exsD缺失株表型特征。【方法】采用overlapPCR技术构建缺失株ΔexsD,PCR检测缺失株ΔexsD的遗传稳定性,比较缺失株与野生株之间的生长速度、泳动能力、胞外酶活性、药物敏感性、毒力,分别采用结晶紫和共聚焦电镜方法测定生物膜的差异变化,通过qRT-PCR方法分析exsD的缺失对T3SS效应蛋白Hop转录的影响。【结果】缺失株ΔexsD构建成功;与野生株相比,ΔexsD遗传稳定性和生长速度无显著差别,但泳动能力极显著上升(P <0.01),胞外蛋白酶活性显著上升(P <0.05),形成生物膜能力在24 h时提高(P <0.05);相比野生株,ΔexsD对米诺环素、庆大霉素、卡那霉素、多西环素、新霉素的敏感性从耐药变成中度敏感;ΔexsD缺失株的毒力显著上升,野生株对斑马鱼的半数致死剂量是缺失株的3.68倍。实时荧光定量结果显示,与HY9901野生型相比,exsD基因的缺失增加了效应蛋白Hop的转录表达。【结论】exsD基因对T3SS的致病性起负调控作用,为进一步阐明exsD基因在溶藻弧菌中的作用机理提供一定的理论基础。

关键词(KeyWords)： 溶藻弧菌;Ⅲ型分泌系统;exsD基因;基因敲除;缺失株;表型特征

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金（32073015）;; 广东省自然科学基金（2021A1515011078）;; 广东省科技创新战略专项资金（大学生科技创新培育）（pdjh2021b0239）;; 广东海洋大学大学生创新创业训练计划项目（CXXL2020131）;; 广东海洋大学本科生创新团队（CCTD201802）

作者(Author): 王俊霖,招茵,苏茵茵,周诗慧,曾福源,谢妙,王娜,简纪常,庞欢瑛

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