锦鲤疱疹病毒-CJ株ORF25基因的克隆及生物信息学分析

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

关键词:  锦鲤疱疹病毒  ORF25基因  生物信息学分析

Key words:  Kio herpesvirus  ORF25 gene  bioinformatics analysis

摘要:

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,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1824bp的ORF25基因,编码608个氨基酸；预测ORF25基因的理论分子质量为66825.44Da,等电点为7.947；疏水性为-3.067～3.056；抗原表位预测显示抗原性良好；结构预测显示,可能存在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。经分析表明,该基因的表达产物可能具有较好的免疫原性。

Abstract:

To study its 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 the ORF25 gene was analyzed by means of bioinformatics softwares. Then the phylogenetic tree was constructed. The results showed that a length of the ORF25 gene, encoding608 aa, was 1824bp. The molecular weight was 66825.44Da and the isoelectric point was 7.947. The protein has a good hydrophobic. The antigenicity of ORF25 was good. According to the protein structure analysis, there may be have 6 N-glycosylation sites, 25 O-glycosylation site and 28 phosphorylation sites. Phylogenetic analysis showed that the ORF25 gene of KHV-CJ was in the same branch with the USA strains. Above all, the ORF25gene may have good immunogenicity.