锦鲤疱疹病毒丹东株ORF83基因克隆及生物信息学分析

发布时间：2018-06-12 13:15

本文选题：锦鲤疱疹病毒 + ORF基因　；参考：《基因组学与应用生物学》2017年04期

【摘要】：锦鲤疱疹病毒病是具有高致病性、传染性的疾病,对渔业生产造成巨大的危害。为了研究锦鲤疱疹病毒衣壳蛋白ORF83的结构和功能,本研究采集感染锦鲤疱疹病毒活鱼样品,经DNAStar软件预测分析、DNA提取,PCR扩增、重组质粒构建等步骤成功克隆到ORF83(1)和ORF83(2)基因。应用生物信息学分析蛋白结构与功能,并构建系统进化树。结果显示,两蛋白均能编码前体蛋白,均由α螺旋、β折叠、无规则卷曲组成,均有4个跨膜区域;抗原性较好;但不含保守结构域;ORF83(1)N端11个氨基酸序列为信号肽序列;ORF83(2)N端33个氨基酸序列为信号肽序列;ORF83(1)氨基酸序列存在1个潜在的N-糖基化位点、ORF83(2)氨基酸序列存在3个潜在的N-糖基化位点,两蛋白均潜在13个磷酸化位点,无潜在的O-糖基化位点;系统进化树分析表明,KHV-DD ORF83与美国株、以色列株、日本株均具有同源性。通过研究表明,本研究成功克隆到ORF83基因,并对其结构和功能进行分析,获得重要的生物信息学数据,为进一步研究锦鲤疱疹病毒发病机制和疱疹病毒抗体制备等提供了理论基础。
[Abstract]:Koi herpesvirus disease is a highly pathogenic and infectious disease, which causes great harm to fishery production. In order to study the structure and function of Koi herpesvirus capsid protein ORF83, live fish samples infected with Koi herpesvirus were successfully cloned into ORF83P1and ORF83m-2) genes by DNA Star software prediction analysis, PCR amplification of DNA extraction and construction of recombinant plasmid. The structure and function of protein were analyzed by bioinformatics and phylogenetic tree was constructed. The results showed that both proteins could encode precursor proteins, which were composed of 伪 -helix, 尾 -fold, irregular crimp, and had four transmembrane regions, and had good antigenicity. However, there were one potential N-glycosylation site and three potential N-glycosylation sites in the N-terminal 11 amino acid sequences with no conserved domain ORF83N terminal and the signal peptide sequence ORF83Amino 1), and the ORF83Amino acid sequence with the N terminal 33 amino acid sequences of ORF83Amins was a signal peptide sequence, and there were three potential N-glycosylation sites in the amino acid sequence of ORF83Amino 1), and there were three potential N-glycosylation sites in the amino acid sequence. The phylogenetic tree analysis showed that KHV-DD ORF83 had homology with American, Israeli and Japanese strains. The results showed that ORF83 gene was cloned successfully, and its structure and function were analyzed, and important bioinformatics data were obtained. It provides a theoretical basis for further study on pathogenesis of Koi herpesvirus and preparation of HSV antibody.
【作者单位】：中国水产科学研究院黑龙江水产研究所;
【基金】：国家科技支撑计划(2012BAD26B01) 现代农业产业技术体系建设专项资金(CARS-46-02) 中央级公益性科研院所基本科研业务费专项(HSY201401)共同资助
【分类号】：S941.41

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