锦鲤疱疹病毒-CJ株ORF136基因的克隆及生物信息学分析  

Cloning and bioinformatics analysis of the ORF136 gene of Koi herpesvirus-CJ strain

作　　者：刘春杰 杨丽华 王好[2] 周井祥[2] 

机构地区：[1]长春科技学院,长春130600 [2]吉林农业大学,长春130118

出　　处：《黑龙江畜牧兽医》2015年第6期172-174,290,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目(30671621)

摘　　要：为了了解锦鲤疱疹病毒中国吉林株(KHV-CJ)ORF136蛋白的结构特征和进化关系,试验用锦鲤尾鳍原代细胞增殖KHV-CJ,提取DNA,经PCR扩增获得ORF136基因,将其克隆到p MD18-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 ORF136基因的结构和功能,并与Gen Bank上已公布的KHV进行比较。结果表明:ORF136有2个跨膜区;抗原表位预测显示,抗原性良好;结构预测显示,可能存在1个潜在的N-糖基化位点和19个潜在的O-糖基化位点;BLAST比对分析显示,KHV-CJ ORF136与KHV-U ORF136和KHV-I ORF136的亲缘关系最近,与KHV-GZ11ORF136的亲缘关系较近,与KHV-GZ10 ORF136、KHV-QY08 ORF136和KHV-J ORF136亲缘关系最远,KHV-CJ可能来源于欧美国家和以色列。说明ORF136基因可能具有较好的免疫原性。To understand the protein structural features and evolutionary relationships of the ORFI36 of the China Jilin strain from Koi herpesvirus （ KHV - CJ）, the primary cells of koi caudal fin were used for the proliferation of the KHV - CJ to extract its DNA. The ORF136 gene was obtained by PCR amplification, and then was cloned into the vector pMD18 -T to constrUct the recombinant plasmid. The structure and function of the ORF136 gene of the KHV - CJ were preliminarily studied by bioinformatics method ,and then compared with those of the other KHV strains published in GenBank. The resuhs showed that the ORF136 gene had two transmembrane regions; the epitope prediction showed that the antigenicity of the ORF136 gene was good; the structure prediction showed that there might be one N -glycosylation site,nineteen potential O - glycosylation sites; A BLAST alignment analysis showed that the KHV -CJ ORF136 gene had the closest kinship with the KHV -U ORF136 gene and KHV - I ORF136 gene, and a closer kinship with the KHV - GZll ORF136 gene, and the farthest kinship with the KHV - GZIO ORF136 gene, KHV- QY08ORF136 gene and KHV- JORF136 gene; the KHV- CJ might come from the European and American countries and Israel. The results indicate that the ORF136 gene may have good immunogenicity.

关 键 词：锦鲤疱疹病毒(KHV) ORF136基因 生物信息学分析 克隆 免疫原性 吉林株 

分 类 号：S852.659.1[农业科学—基础兽医学] S965.116[农业科学—兽医学]