首页 | 本学科首页   官方微博 | 高级检索  
     检索      

鲤疱疹病毒3型T分离株主要免疫原性蛋白的鉴定
引用本文:石存斌,任燕,曾伟伟,王英英,李莹莹,王庆,郑树城.鲤疱疹病毒3型T分离株主要免疫原性蛋白的鉴定[J].淡水渔业,2018(2):68-72.
作者姓名:石存斌  任燕  曾伟伟  王英英  李莹莹  王庆  郑树城
作者单位:1. 中国水产科学研究院珠江水产研究所,农业部渔药创制重点实验室,广东省水产动物免疫技术重点实验室,广州510380;2. 上海海洋大学水产与生命学院,上海201306
基金项目:中国水产科学研究院基本科研业务费资助,国家科技支撑计划,现代农业产业技术体系建设专项资金资助,中国水产科学研究院基本科研业务费资助
摘    要:为了鉴定鲤疱疹病毒3型(Cyprinid Herpesvirus 3,CyHV-3)主要免疫原性蛋白,研究采用CyHV-3-T分离株感染CCB细胞系,运用蔗糖密度梯度超速离心方法对CyHV-3进行纯化,纯化的病毒颗粒经SDSPAGE和考马斯亮蓝染色后,用锦鲤(Cyprinus carpio haematopterus)抗CyHV-3阳性血清进行Western blotting分析和液相色谱串联质谱鉴定。结果表明,透射电镜下可观察到大量完整囊膜包裹或只有裸露核衣壳的CyHV-3颗粒,Western blotting结果显示抗CyHV-3阳性血清与多种病毒蛋白具有明显特异性免疫反应,质谱鉴定表明其中4种免疫原性蛋白分别为ORF92、ORF66、ORF72和ORF81,其中ORF66和ORF72为首次鉴定的具有免疫原性衣壳蛋白。本研究将为CyHV-3血清学诊断方法的建立、亚单位疫苗或DNA疫苗的研制提供更多候选抗原。

关 键 词:鲤疱疹病毒3型  锦鲤疱疹病毒  免疫原性  免疫蛋白质组学  衣壳蛋白

Identification of major immunogenic proteins from Cyprinid Herpesvirus 3 strain T
SHI Cun-bin,REN Yan,ZENG Wei-wei,WANG Ying-ying,LI Ying-ying,WANG Qing,ZHENG Shu-cheng.Identification of major immunogenic proteins from Cyprinid Herpesvirus 3 strain T[J].Freshwater Fisheries,2018(2):68-72.
Authors:SHI Cun-bin  REN Yan  ZENG Wei-wei  WANG Ying-ying  LI Ying-ying  WANG Qing  ZHENG Shu-cheng
Abstract:CCB cell line was infected with CyHV-3 isolate T in this study to identify major immunogenic proteins from CyHV-3.Virus suspensions were collected after freezing and thawing repeatedly when apparent cytopathic effect (CPE) was observed and purification was performed by 20% to 66% sucrose density gradient in combination with ultracentrifuga-tion.The obtained purified virions were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS -PAGE) and stained with coomassie brilliant blue.Furthermore, western blotting analysis was performed by using anti-ser-um from koi naturally infected with CyHV-3.Some protein bands showed clearly and specifically immunoreactive, they were cut for further characterization through liquid chromatography tandem mass spectrometry.Transmission electron microscope (TEM) results showed that the purified CyHV-3 virions contained intact envelope or naked nucleocapsid with high purity and typical characteristic in line with previous report.Quite some protein bands showed immunoreactive using anti-serum against CyHV-3 from koi by western blotting, 4 of which clearly immunogenic proteins were further identified as capsid protein ORF92, ORF66, ORF72 and envelope protein ORF81 through LC-MS/MS, respectively.In addition, ORF66 and ORF72 was also the newly identified capsid protein showed immunogenicity.This study might provide more antigen can-didates for development of serological diagnosis methods for CyHV-3 and DNA vaccine or subunit vaccine preparation.
Keywords:
本文献已被 CNKI 万方数据 等数据库收录!
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号