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副溶血弧菌(Vibrioparahaemolyticus)tdh2基因和鳗弧菌(Kanguillarum)ompU基因二联DNA疫苗制备及其对大菱鲆(Scophthalmusmaximus)免疫保护作用
引用本文:刘 瑞,赵明君,杨 慧,陈吉祥.副溶血弧菌(Vibrioparahaemolyticus)tdh2基因和鳗弧菌(Kanguillarum)ompU基因二联DNA疫苗制备及其对大菱鲆(Scophthalmusmaximus)免疫保护作用[J].海洋与湖沼,2011,42(4):580-586.
作者姓名:刘 瑞  赵明君  杨 慧  陈吉祥
作者单位:中国海洋大学海洋生物遗传学与基因资源利用教育部重点实验室;中国海洋大学海洋生物遗传学与基因资源利用教育部重点实验室;西北工业大学生命学院;中国海洋大学海洋生物遗传学与基因资源利用教育部重点实验室;兰州理工大学石油化工学院
基金项目:国家自然科学基金项目, 30972275 号; 国家 863 计划项目, 2007AA09Z416 号; 农业部公益性行业专项, nyhyzx07-046 号
摘    要:采用基因工程的方法,将副溶血弧菌的热稳定直接溶血素基因tdh2和鳗弧菌外膜蛋白基因ompU进行融合,在大肠杆菌中得到表达,并利用该融合基因构建二联DNA疫苗pEGFP.N1/tdh2-ompU。用DNA疫苗按10和50μg/尾的剂量通过肌肉注射免疫大菱鲆,对大菱鲆抵抗致病性副溶血弧菌和鳗弧菌的免疫效果进行研究。结果表明...

关 键 词:副溶血弧菌  鳗弧菌  热稳定溶血素  外膜蛋白  二联DNA疫苗  大菱鲆  免疫性
收稿时间:8/4/2010 12:00:00 AM
修稿时间:2010/10/9 0:00:00

CONSTRUCTION AND PROTECTION EVALUATION OF A BIVALENT DNA VACCINE CONTAINING tdh2 OF VIBRIO PARAHAEMOLYTICUS AND ompU OF V. ANGUILLARUM ON TURBOT SCOPHTHALMUS MAXIMUS
LIU Rui,ZHAO Ming-Jun,YANG Hui and CHEN Ji-Xiang.CONSTRUCTION AND PROTECTION EVALUATION OF A BIVALENT DNA VACCINE CONTAINING tdh2 OF VIBRIO PARAHAEMOLYTICUS AND ompU OF V. ANGUILLARUM ON TURBOT SCOPHTHALMUS MAXIMUS[J].Oceanologia Et Limnologia Sinica,2011,42(4):580-586.
Authors:LIU Rui  ZHAO Ming-Jun  YANG Hui and CHEN Ji-Xiang
Institution:Key Laboratory of Marine Genetics and Breeding, Ministry of Education, Ocean University of China;Key Laboratory of Marine Genetics and Breeding, Ministry of Education, Ocean University of China;Faculty of Sciences, Northwestern Polytechnical University;Key Laboratory of Marine Genetics and Breeding, Ministry of Education, Ocean University of China;School of Petrochemical Engineering, Lanzhou University of Technology
Abstract:Vibriosis is one of the most prevalent fish diseases caused by bacteria that belong to the genus Vibrio, in which Vibrio parahaemolyticus and V. anguillarum are two most common pathogenic agents. In this study, a thermostable direct hemolysin (TDH) gene tdh2 of V. parahaemolyticus and an outer membrane protein (OMP) gene ompU of V. anguillarum were inserted into prokaryotic expression vector pET28a (+) successively. Then the gene encoding the fusion protein TDH2-OmpU was used to construct bivalent DNA vaccine. The protective efficiency of such vaccine against V. parahaemolyticus and V. anguillarum was evaluated in turbot Scophthalmus maximus. Fish immunized with a single intramuscular injection of different doses of the tdh2-ompU DNA vaccine showed significantly higher serum antibody levels after vaccination, compared to fish injected with the control eukaryotic expression vector pEGFP-N1. Four weeks after inoculation, turbot vaccinated with the bivalent DNA vaccine were exposed to V. parahaemolyticus and V. anguillarum by intraperitoneal injection and intramuscular injection, respectively. The highest relative percent survival (RPS) rates, 100% against V. parahaemolyticus and 35% against V. anguillarum, respectively, were recorded. Furthermore, humoral immune responses and protective effects significantly increased at higher vaccine doses.
Keywords:Vibrio parahaemolyticus  V  anguillarum    Thermostable direct hemolysin    Outer membrane protein  Bivalent DNA vaccine    Turbot Scophthalmus maximus    Immunity
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