鸭瘟病毒UL28基因的克隆、鉴定及序列分析  

Cloning,Characterization and Sequence Analysis of Duck plague virus UL28 Gene

作　　者：娄昆鹏[1] 程安春[1,2] 汪铭书[1,2] 朱德康[1,2] 贾仁勇[1] 罗启慧[1] 刘菲[2] 陈孝跃[1] 

机构地区：[1]四川农业大学动物医学院禽病防治研究中心,雅安625014 [2]动物疫病与人类健康四川省重点实验室,雅安625014

出　　处：《农业生物技术学报》2010年第4期737-745,共9页Journal of Agricultural Biotechnology

基　　金：教育部"长江学者和创新团队发展计划"创新团队项目(No.PCSIRT0848);国家自然科学基金项目(No.30771598);现代农业产业技术体系建设专项资金资助(No.nycytx-45-12);高等学校科技创新工程重大项目培育资金项目(No.706050)共同资助

摘　　要：对已构建的鸭瘟病毒(DPV)DNA基因文库重组质粒序列测序后,结合ORF Finder和BLAST工具分析得到了DPVUL28基因的开放阅读框。PCR扩增后将其连接至pMD18-T克隆载体,获得完整基因。经测序、PCR和双酶切鉴定,进一步用核酸探针斑点杂交确认完整片段(ORF2412)属于DPVDNA(GenBank登录号EF643557)。运用生物信息学分析软件NNPPversion2.2、Translate tool、DNAStar等分析序列特性,并用EMBOSS软件包的CHIPS和CUSP程序分析密码子使用情况。结果显示,其具有疱疹病毒_UL28类似家族成员的典型特征,包含1个保守结构域:疱疹病毒加工和组装蛋白(PRTP),并含有多个与功能相关的磷酸化和N-糖基化位点,编码多肽疏水区大于亲水区,是一种膜外蛋白。DPVUL28基因与禽类疱疹病毒(α-疱疹病毒)的亲缘关系最近。UL28基因在密码子选择使用上显示出较强的偏爱性。A new gene （UL28） from Duck plaque virus （DPV） DNA gene library constructed in Key Laboratory of Animal Disease and Human Health of Sichuan Province was selected and sequenced.A complete open reading frame （ORF2412） was found by ORF Finder and BLAST.It was amplified by PCR and cloned into pMD18-T vector.The positive plasmid was selected and identified by PCR and incision enzyme.Furthermore,it was confirmed by nucleic acid dot-hybridization to make sure the ORF2412 belonged to DPV DNA,and analyzed by bioinformatics tools of NNPP version 2.2,Translate tool,DNAStar and so on.Furthermore,CHIPS and CUSP methods of EMBOSS package were used to analyze the codon usage.The results showed that the typical characteristic of herpesvirus_UL28 was possessed;it contained one structural domain of PRTP and numbers of feature associated with the phosphorylation sites and N-glycosylation sites.Hydrophobic region was greater than hydrophilic region;it was a kind of protein out of membrane.Phylogenetic demonstrated that it was more closely related to poultry herpesvirus （α-herpesvirus）.In addition,DPV UL28 gene was shown strong bias in codon usage.

关 键 词：鸭瘟病毒 UL28基因 克隆 序列分析 

分 类 号：S858.325.3[农业科学—临床兽医学]