1株J亚群禽白血病病毒env基因克隆及生物信息学分析  

Cloning and Bioinformatics Analysis of env Gene of Avian Leukosis Virus Subgroup J Strain

作　　者：陈玫婷 郑从森 梁泽贤 林巧儿 常传哲 周俊 冯军 李林林 覃丽梅 CHEN Meiting;ZHENG Congsen;LIANG Zexian;LIN Qiaoer;CHANG Chuanzhe;ZHOU Jun;FENG Jun;LI Linlin;QIN Limei(College of Life Science and Engineering,Foshan University,Foshan 528231,China;Institute of Animal Health,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,China)

机构地区：[1]佛山科学技术学院生命科学与工程学院,佛山528231 [2]广东省农业科学院动物卫生研究所,广州510640

出　　处：《中国畜牧兽医》2023年第5期1785-1795,共11页China Animal Husbandry & Veterinary Medicine

基　　金：广东省自然科学基金面上项目(2021A1515012388)。

摘　　要：【目的】通过生物信息学方法分析1株禽白血病病毒(Avian Leukosis virus,ALV)的囊膜蛋白env的分子特征。【方法】利用鸡成纤维细胞(DF-1)培养疑似禽白血病病鸡的血清样品,通过ELISA和实时荧光定量PCR鉴定ALV亚群。对env基因PCR产物进行测序,利用SeqMan拼接env基因。将该env基因与国内外多个ALV亚群进行序列比对及遗传进化分析,并借助多种生物信息学软件对env蛋白进行分析,包括理化性质、亲/疏水性、跨膜区、信号肽、糖基化位点、磷酸化位点、乙酰化位点、二级结构、三级结构、抗原决定簇和B细胞抗原表位预测。【结果】ALV-J毒株的env基因全长1719 bp,获得GenBank登录号为OP837418,其与J亚群ALV位于同一进化分支,相似性为89.3%~94.8%。该env蛋白有2个跨膜区,是由572个氨基酸组成的不稳定的亲水蛋白,不稳定系数预测值为43.49,总平均亲水性为―0.201;该env蛋白为非分泌蛋白,无信号肽,有17个N-糖基化修饰位点、77个O-糖基化修饰位点、42个磷酸化修饰位点、21个潜在抗原决定簇和9个连续碱基数超过10个的B细胞线性表位。该env蛋白的二级结构中无规则卷曲占比最大,为38.29%。【结论】env基因是导致ALV遗传变异的关键基因,其编码的囊膜蛋白env具有不稳定性,存在多个蛋白修饰位点和抗原表位。【Objective】The purpose of this study was to characterize the envelope protein(env)of an Avian leukosis virus(ALV)strain by bioinformatics method.【Method】The serum samples from suspected avian leukosis ckickens were cultured in chicken fibroblast(DF-1),and the subgroups of ALV was identified by ELISA and Real-time quantitative PCR.The PCR products of env gene were sequenced and spliced by SeqMan.The env gene sequence was compared with several ALV subpopulations both domestically and abroad,and the genetic evolution analysis was performed.The env proteins were analyzed with a variety of bioinformatics software,including physicochemical properties,hydrophilicity and hydrophobicity,transmembrane domain,signal peptide,glycosylation site,phosphorylation site,secondary structure,tertiary structure,acetylation site,epitope and B cell epitopes prediction.【Result】The full length of env gene of ALV-J strain was 1719 bp,and GenBank accession No.:OP837418.It was located in the same evolutionary branch with ALV-J subgroup,and the similarity was 89.3%-94.8%.The env protein had two transmembrane domains and was a hydrophilic and unstable protein composed of 572 amino acids with a predicted instability coefficient of 43.49 and a total average hydrophilicity of-0.201.The env protein was a non-secreted protein,without signal peptide,with 17 N-glycosylation modification sites,77 O-glycosylation modification sites,42 phosphorylation modification sites,21 potential epitope sites and 9 linear B cell epitopes with more than 10 consecutive bases.In the secondary structure of the env protein,the proportion of randorn coil was the largest(38.29%).【Conclusion】The env gene was a key gene that caused genetic variation in ALV,and its encoded envelope protein env was unstable,with multiple protein modification sites and antigenic epitopes.

关 键 词：禽白血病病毒(ALV) J亚群 ENV基因 生物信息学 

分 类 号：S858.31[农业科学—临床兽医学]