锦鲤疱疹病毒-CJ株ORF25基因的克隆及生物信息学分析

为了解锦鲤疱疹病毒中国吉林株(KHV-CJ)ORF25蛋白的结构特征和进化关系,采用框镜鲤尾鳍原代细胞增殖KHV-CJ,提取其DNA,经PCR扩增,获得ORF25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 ORF25基因的结构和功能,并与GenBank上已公布的三株KHV构建系统进化树。结果显示,获得了长1824 bp的ORF25基因,编码608个氨基酸;预测ORF25基因编码蛋白的理论分子质量为66825.44 Da,等电点为7.947;抗原表位预测显示抗原性良好;编码蛋白存在6个N-糖基化位点、25个O-糖基化位点和28个磷酸化位点;系统进化树分析显示,与锦鲤疱疹病毒美国株属同一分支。

Cloning and bioinformatics analysis of ORF25 gene of Kio herpesvirus-CJ strain

To study structure and function,the ORF25 gene was amplified by PCR from koi herpesvirus China’s Jilin strains(KHV-CJ) infected mirror carp primary fin cells,and then was cloned into the vector pMD18-T to construct the recombinant plasmid pMD18-T-ORF25.The structure and function of ORF25 gene was analyzed by means of bioinformatics softwares.Then the phylogenetic tree was constructed.The results showed that the length of ORF25 gene,encoding 608 amino acids,was 1824 bp.The molecular weight was 66825.44 Da and the isoelectric point was 7.947.The protein has a good hydrophobicity.The antigenicity of ORF25 was good.According to the protein structure analysis,there may be 6 N-glycosylation sites,25 O-glycosylation sites and 28 phosphorylation sites.Phylogenetic analysis showed that the ORF25 gene of KHV-CJ was in the same branch with the USA strains.