锦鲤疱疹病毒CJ株ORF83基因的克隆及生物信息学分析  

Cloning and bioinformatics analysis of ORF83 gene of kio herpesvirus CJ strain

作　　者：李新伟[1] 周井祥[2] 朱霞[1] 王好[1] 吕文亮[1] 

机构地区：[1]吉林农业大学动物生产及产品质量安全教育部重点实验室,吉林长春130118 [2]吉林农业大学动物科技学院,吉林长春130118

出　　处：《中国兽医科学》2011年第10期1016-1020,共5页Chinese Veterinary Science

基　　金：国家自然科学基金项目(30671621)

摘　　要：为了解锦鲤疱疹病毒中国吉林株(KHV-CJ)ORF83蛋白的结构特征和进化关系,用框镜鲤尾鳍原代细胞增殖KHV-CJ,提取其DNA,经PCR扩增,获得ORF83基因,将其克隆到pMD18-T载体中,构建重组质粒。应用生物信息学方法初步分析KHV-CJ ORF83基因的结构和功能,并与GenBank上已公布的3株KHV构建系统进化树。结果显示,获得了长687 bp的ORF83基因,编码229个氨基酸;编码蛋白的理论分子质量为25 822.18 u,等电点为8.600;疏水性为-2.733～3.100;信号肽切割部位最可能位于第22位的G(甘氨酸)和第23位的V(缬氨酸)之间;跨膜结构分析表明,ORF83有4个跨膜区;抗原表位预测显示,编码蛋白的抗原性良好;结构预测显示,ORF83可能存在1个N-糖基化位点、11个O-糖基化位点和13个磷酸化位点;系统进化树分析显示,KHV-CJ株与锦鲤疱疹病毒美国株和以色列株属同一分支。结果表明,ORF83基因编码的蛋白可能具有较好的免疫原性,能够诱导产生中和抗体。The ORF83 gene was cloned by PCR method from kio herpesvirus China Jilin（KHV-CJ） strain infected Mirror carp fin cells,and was then cloned into the pMD18-T vector to construct a recombinant plasmid.The structure and function of the gene was researched with bioinformatics method.Phylogenetic tree was constructed with other ORF83 genes from three KHV strains in GenBank.In result,an ORF83 gene of 687bp was obtained from KHV-CJ strain.The ORF83 gene encoded a protein of 229aa with a molecular weight of 25822u,and isoelectric point as 8.600,and the hydrophobicity was between-2.733 and 3.100.The most probably signal peptide splitting site was between 22 bit（glycine） and 23 bit（Valine）.The coding protein of ORF83 had four transmembrane regions and fine antigenic determinants showed by online analysis.Structure analysis showed that there might be one N-glycosylation sites,11 O-glycosylation site and 13 phosphorylation sites in the encoding protein.Phylogenetic analysis showed that KHV-CJ strain was at the same branch with strains of the United States and the Israel.Results showed that the protein encoded by the ORF83 gene from KHV-CJ strain might have fine immunogenicity,and could induce neutralizing antibodies.

关 键 词：锦鲤疱疹病毒 ORF83基因 生物信息学分析 

分 类 号：S852.659.1[农业科学—基础兽医学]