鸭瘟病毒UL47基因克隆及其分子特性分析  

Cloning and molecular characterization of UL47 gene of duck plague virus

作　　者：罗丹丹[1] 程安春[1,2] 汪铭书[1,2] 沈爱梅[1] 朱德康[1,2] 贾仁勇[2] 罗启慧[2] 崔恒敏[2] 王印[2] 徐志文[2] 王小玉[2] 陈孝跃[1,2] 

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

出　　处：《中国兽医学报》2011年第1期29-35,共7页Chinese Journal of Veterinary Science

基　　金：教育部“长江学者和创新团队发展计划”创新团队项目(IRT0848),教育部高等学校科技创新工程重大项目(706050);现代农业产业技术体系建设专项资金资助项目(nycytx-45-42)

摘　　要：通过测定本实验室构建的鸭瘟病毒(Duck plague virus,DPV)DNA基因文库中重组质粒的DNA序列,结合NCBI的ORF Finder和Blast工具分析得到了该病毒UL47基因的ORF。采用PCR扩增出了UL47基因并将其克隆到pMD18-T载体上,经PCR和酶切鉴定以及进一步的核酸斑点杂交试验证实该基因即为DPV UL47基因。利用生物信息学软件ProtScale、SignalP3.0、Scansite、TMpred、Prosite、DNAStar以及在线的EMBOSS等分析了UL47基因的分子特性。结果显示,该基因大小为2 367bp,编码788aa,而且与GenBank上多种α疱疹病毒同源蛋白的核酸和氨基酸序列具有较高的同源性。系统进化树分析表明,DPV UL47与禽类疱疹病毒(α-疱疹病毒)的进化关系最近。密码子偏爱性结果显示,DPV UL47编码相同氨基酸的不同密码子使用频率差异较大,UL47基因密码子使用模式更接近真核生物。研究结果为进一步开展DPV UL47基因功能研究奠定了基础。Sequencing a recombinant plasmid selected from DPV CHv strain genomic library constructed in our lab,and we got the UL47 gene by using Blast and ORF Finder on NCBI.DPV UL47 gene was amplified by PCR and cloned into pMD18-T vector,and strongly confirmed by PCR amplification,restriction digestion and oligonucleotide probe hybridization.Then the DPV UL47 was analyzed by bioinformatics tools of ProtScale,SignalP3.0,Scansite,TMpred,Prosite,DNAStar and EMBOSS online programs.The results showed that the DPV UL47 gene was composed of 2 367 nucleotides,and encoding a polypeptide of 788 amino acid residues.Moreover,the nucleic acid and amino acid sequence of DPV UL47 had higher homology with its homologous protein of alphaherpesvirus than those of others.The phylogenetic tree analysis showed that the DPV clusters with some avian herpesviruses such as GaHV-2,GaHV-3,MeHV-1 and MDV-2 in a monophyletic clade are closer than those of other alphaherpesvirus members and as a result it has a close evolutionary relationship with alphaherpesviruses.Besides,condon preference analysis demonstrated that the alternative codons for the same amino acid in UL47 had distinctly different frequency and comparison of the codon usage in the UL47 gene of different organisms revealed that there were 30 codons showing distinct usage differences between DEV and Escherichia coli,equivalently 24 between DEV-to-yeast and DEV-to-human.Therefore,the eukaryotic expression system may be more suitable for the expression of the DEV UL47 gene.

关 键 词：鸭瘟病毒 UL47基因 分子特性 

分 类 号：S852.65[农业科学—基础兽医学]