摘要:
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720bp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26 107.1Da,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。
Abstract:
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 ORF51was 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 239amino acids,with 100%identity with AngHV-1ORF51gene.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 1N-glycosylation sites,1O-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.
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