摘要
鲫(Carassius auratus)造血器官坏死症是我国养殖鲫中新出现的一种传染性病毒性疾病,其病原为鲤疱疹病毒Ⅱ型(Cy HV-2),为研制针对该病的有效诊断试剂和疫苗,分析了Cy HV-2 ORF25基因编码蛋白的抗原表位,利用原核表达系统对Cy HV-2 ORF25编码蛋白富含抗原表位的区域进行截短表达,制备了抗Cy HV-2 ORF25编码蛋白的多克隆抗体,通过间接免疫荧光和免疫保护力测定等实验研究了截短表达蛋白的免疫原性。结果显示,原核表达的截短蛋白t ORF25分子质量约为28 k W,蛋白纯化后免疫日本大耳兔制备的多克隆抗体可特异性识别在Gi CB上增殖的Cy HV-2。将截短表达蛋白t ORF25免疫异育银鲫后,感染Cy HV-2,其相对免疫保护率达47%。
Cyprinid herpesvirusⅡ(CyHV-2)infection is a newly emerged infectious disease of farmed Carassius auratus in China and has caused huge economic losses to the aquaculture industry.Currently,the researches on the diagnosis and control methods of this disease have become a great concern.In this study,the epitopes sites of CyHV-2 ORF25 encoding protein were analyzed.The antigen-rich region of this gene was truncated and expressed in prokaryotic expression system.Meanwhile,the polyclonal antibody against the expressed protein was generated and the immunogenicity of the trucated CyHV-2 ORF25 protein was investigated by indirect immunological fluorescent assay and the immune protection test.The results showed that the truncated protein,tORF25,had a molecular weight of 28 kW,which was the same as the predicted size.Ployclonal antibody was prepared in Japanese white rabbit by immunizing the animal with purified tORF25 protein.The indirect fluorescent assay demonstrated that this polyclonal antibody could specifically recognize CyHV-2 proliferated on the GiCB cells.The challenge test showed that the relative percentage survival of gibel carp immunized with the tORF25 protein reached to 47%.
作者
周勇
史玉恒
范玉顶
刘文枝
江南
赵建青
许晨
曾令兵
ZHOU Yong;SHI Yu-heng;FAN Yu-ding;LIU Wen-zhi;JIANG Nan;ZHAO Jian-qing;XU Chen;ZENG Ling-bing(Yangtze River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Wuhan 430223,China)
出处
《淡水渔业》
CSCD
北大核心
2018年第6期40-45,共6页
Freshwater Fisheries
基金
中国水产科学研究院中央级公益性科研院所基本科研业务费专项(2017HY-ZD0306
2016HY-ZD0501)
国家自然科学基金(31802346)
国家"现代农业产业技术体系建设专项资金"资助(CARS-45-16)
武汉市关键技术攻关计划(2016020101010083)
国家质检总局科技计划(2016IK116)
关键词
鲤疱疹病毒Ⅱ型
ORF25截短蛋白
原核表达
多克隆抗体
免疫原性
Cyprinid herpesvirusⅡ
truncated ORF25 protein
prokaryotically expression
polyclonal antibody
immunogenicity
