鳗鲡疱疹病毒FJ株ORF51基因的克隆及生物信息学分析

为了解本课题组分离的鳗鲡疱疹病毒福建株（AngHV-FJ） ORF51的结构特征，扩繁了 AngHV-FJ ，提取其基因组DNA ，经PCR扩增，获得ORF51基因，将其克隆至pMD19-T载体中，进行DNA测序，用生物信息学方法分析AngHV-FJ ORF51基因及其编码蛋白的结构和功能。结果显示，扩增到长约720 bp的ORF51基因序列，其与GeneBank上鳗鲡疱疹病毒欧洲株（AngHV-1） ORF51基因的序列完全一致。该基因编码239个氨基酸；预测ORF51基因编码蛋白的分子量为26107.1 Da ，等电点为7.66，是疏水性蛋白且偏碱性；有4个跨膜域；抗原表位预测显示抗原性较好；结构预测显示，存在1个N-糖基化位点、1个O-糖基化位点、10个磷酸化位点；亚细胞定位预测显示其主要存在于内质网。本研究为解析ORF51的功能及开展AngHV的基因工程疫苗研究奠定了基础。

Cloning and Bioinformatics Analysis of ORF51 Gene of Anguillae Herpesvirus FJ Strain

Anguillae herpesvirus Fujian strain （AngHV-FJ） was first isolated from China mainland by our lab .To study the structure and function of AngHV-FJ ORF51 ,AngHV-FJ was propagated ,and the ORF51 gene was amplified by PCR from the extracted virus genomic DNA ,then ORF51 was cloned into the vector pMD19-T for sequencing .The nucleotide and protein sequence of ORF51 was analyzed by bioinformatics softwares .The ORF51 gene is 720bp in length ,encoding 239 amino acids ,with 100% identity with AngHV-1 ORF51 gene .The protein has 26107.1Da molecular weight ,7.66 isoelectric point ,hydrophobic and alkaline ,four transmembrane regions with high antigenicity .Structure analysis indicated that ,the protein has 1 N-glycosylation sites ,1 O-glycosylation sites and 10 phosphorylation sites .Subcellular localization prediction showed that the protein is mainly localized in the endoplasmic reticulum . The study provided a reference for the future ORF51 prokaryotic or eukaryotic expression , and established the foundation for the Anguillae herpesvirus research on genetic background information ,pathogenesis ,molecular epidemiology and genetic engineering vaccine development .