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ISSN 1000-0615

主管 中国科学技术协会

主办 中国水产学会

鲤疱疹病毒Ⅱ型主要免疫原性蛋白的鉴定

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2020. 鲤疱疹病毒Ⅱ型主要免疫原性蛋白的鉴定. 水产学报, 44(9): 1441-1447. doi: 10.11964/jfc.20190611828
引用本文: 2020. 鲤疱疹病毒Ⅱ型主要免疫原性蛋白的鉴定. 水产学报, 44(9): 1441-1447. doi: 10.11964/jfc.20190611828
Wa GAO, Hong WEN, Hao WANG, Jiaquan LU, Liqun LÜ, Yousheng JIANG. 2020. Identification of major immunogenic proteins from Cyprinid herpesvirus 2. Journal of Fisheries of China, 44(9): 1441-1447. doi: 10.11964/jfc.20190611828
Citation: Wa GAO, Hong WEN, Hao WANG, Jiaquan LU, Liqun LÜ, Yousheng JIANG. 2020. Identification of major immunogenic proteins from Cyprinid herpesvirus 2. Journal of Fisheries of China, 44(9): 1441-1447. doi: 10.11964/jfc.20190611828

鲤疱疹病毒Ⅱ型主要免疫原性蛋白的鉴定

  • 基金项目:

    国家现代农业产业技术体系建设专项(CARS-45-19);第四届中国科协青年人才托举工程(中国水产学会);上海海洋大学优秀本科生进实验室项目(2018)

详细信息

Identification of major immunogenic proteins from Cyprinid herpesvirus 2

  • Fund Project: 国家现代农业产业技术体系建设专项(CARS-45-19);第四届中国科协青年人才托举工程(中国水产学会);上海海洋大学优秀本科生进实验室项目(2018)
More Information
  • 为了鉴定CyHV-2的主要免疫原性蛋白,本研究用分离的CyHV-2-YC-1分离株(下简称CyHV-2)感染异育银鲫尾鳍细胞系(GiCF),用蔗糖密度梯度超速离心法对细胞感染液中的CyHV-2进行纯化,纯化后的CyHV-2病毒免疫小鼠制备抗CyHV-2多克隆抗体。纯化的病毒颗粒经变性聚丙烯酰胺凝胶电泳 (SDS-PAGE)和考马斯亮蓝染色后,用抗CyHV-2多克隆抗体进行Western blotting分析和质谱鉴定。结果显示,透射电镜下观察发现50%~66%的蔗糖梯度多见完整囊膜包裹的CyHV-2病毒颗粒,也有少量囊膜破损的病毒颗粒。Western blotting结果显示,抗CyHV-2多克隆抗体与多种病毒蛋白具有特异性免疫反应,质谱鉴定显示,其中8种主要免疫原性蛋白分别是ORF92、ORF115、ORF25、ORF57、ORF66、ORF72、ORF131和ORF132。研究表明,通过蔗糖密度梯度超速离心法提纯CyHV-2病毒颗粒后,本研究制备的抗CyHV-2多克隆抗体能够特异性识别CyHV-2病毒的主要免疫原性蛋白。本研究将为CyHV-2免疫学检测方法的建立以及疫苗的研制提供更多的候选抗原。
  • 加载中
  • 图 图版Ⅰ  CyHV-2感染异育银鲫尾鳍细胞

    Figure 图版Ⅰ.  GiCF cells infected with CyHV-2

    图 1  CyHV-2蔗糖密度梯度超速离心纯化结果

    Figure 1.  Results of CyHV-2 purification by sucrose density gradient ultracentrifugation

    图 图版Ⅱ  透射电镜下纯化的CyHV-2颗粒

    Figure 图版Ⅱ.  Purified particles of CyHV-2 under transmission electron microscope

    图 2  纯化的CyHV-2病毒SDS-PAGE电泳(a)和Western blotting结果(b)

    Figure 2.  Results of SDS-PAGE of purified CyHV-2 virions (a) and Western blotting analysis (b)

    表 1  CyHV-2主要免疫原性蛋白LC-MS/MS鉴定

    Table 1.  Identification of major immunogenic proteins from CyHV-2 by LC-MS/MS

    序号
    no.
    蛋白
    protein
    蛋白编号
    accession
    分子质量/ku
    mass
    可信度
    −10lgP
    总肽段总数
    peptides
    定位
    localization
    蛋白描述
    description
    1 ORF92 YP_007003911.1 140 447 67 核衣壳
    nucleocapsid
    主要衣壳蛋白
    major capsid protein
    2 ORF115 YP_007003933.1 84 245 13 囊膜
    envelope
    膜蛋白
    membrane protein
    3 ORF25 YP_007003844.1 66 155 5 囊膜
    envelope
    膜蛋白
    membrane protein
    4 ORF57 YP_007003878.1 64 348 32 未知
    unknown
    未知蛋白
    unknown protein
    5 ORF66 YP_007003885.1 45 309 24 核衣壳
    nucleocapsid
    衣壳三重联体蛋白1
    putative capsid triplex subunit 1
    6 ORF72 YP_007003891.1 41 324 17 核衣壳
    nucleocapsid
    衣壳三重联体蛋白2
    putative capsid triplex subunit 2
    7 ORF131 YP_007003952.1 35 87 2 囊膜
    envelope
    膜蛋白
    membrane protein
    8 ORF132 YP_007003951.1 17 150 4 囊膜
    envelope
    膜蛋白
    membrane protein
    注:蛋白可信度得分,值越大表明该蛋白包含的可信肽段越多
    Notes: −10lgP, a larger value indicates that the protein contains more reliable peptides
    下载: 导出CSV
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收稿日期:  2019-06-06
修回日期:  2019-09-17

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