锦鲤疱疹病毒CJ株ORF27基因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）ORF27基因编码蛋白的结构特征和进化关系，采用镜鲤尾鳍原代细胞增殖KHV-CJ，提取其DNA，经PCR扩增，获得ORF27基因，将其克隆在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 ORF27基因的结构和功能，并与GenBank上公布的三株KHV构建系统进化树。结果显示：获得207bp的ORF27基因，编码69个氨基酸；预测ORF27编码蛋白的理论分子质量为7366.62Da，等电点为4.487；抗原表位预测显示抗原性良好；编码蛋白存在1个N-糖基化位点、4个O糖基化位点和5个磷酸化位点；系统进化树分析表明与锦鲤疱疹病毒美国株（KHV-U）属同一分支。

Cloning and Bioinformatics Analysis of ORF27 Gene in Koi Herpesvirus CJ Strain

The ORF27 gene in China＇ s Jilin strains （KHV-CJ）of koi herpesvirus was amplified from primary fin cells of infected mir-ror carp by PCR, and then was cloned into the vector pMD 18-T to construct the recombinant plasmid pMD 18-T-ORF27. The structure and fimction of ORF27 gene was analyzed by means of bioinformatics software, and then the phylogenetic tree was constructed. The results showed that the ORF27 gene encoding 69 amino acids was 207bp in length, with molecular weight of 7366.62 Da and isoelec-tric point of 4.487. It was found that the ORF27 had good hydrophobicity, and antigenicity. The protein structure analysis revealed that there were 1 N-glycosylation sites, 40-glycosylation sites and 5 phosphorylation sites in the ORF27. Phylogenetic tree showed that the ORF27 gene of KHV-CJ was in the same branch with the USA strains（KHV-U）of koi herpesvirus.