鸭坦布苏病毒E基因的分子特性分析  

The Molecular Property Analysis of Duck Tembusu Virus E Gene

作　　者：孙涛[1] 王超[2] 邓明俊[1] 王群[1] 郑小龙[1] 徐彪[1] 

机构地区：[1]山东出入境检验检疫局,山东青岛266002 [2]泰安出入境检验检疫局,山东泰安250014

出　　处：《中国动物检疫》2015年第12期59-65,68,共8页China Animal Health Inspection

摘　　要：为给鸭坦布苏病毒(duck tembusu virus,DTMUV)囊膜糖蛋白选择良好的抗原域和宿主表达系统提供参考依据,本研究通过选取DTMUV QD株基因文库中的一个重组质粒并测序,结合NCBI的ORF Finder和BLAST工具发现一个全长为1488 bp完整开放阅读框架(Open Reading Frame,ORF)片段。将该ORF编码蛋白通过NCBI的BLASTP分析和DNAstar进化树分析,确定该蛋白与类黄病毒E基因编码的蛋白具有较高的相似性。随后应用生物信息学分析工具Conserved Domains确定保守结构域、运用Signal P4.1预测信号肽、TMHMM 2.0预测跨膜区。应用在线程序Net NGlyc1.0预测糖基化位点,Net Phos2.0预测磷酸化位点、Prot Scale进行疏水性分析。最后运用在线EMBOSS和自动蛋白同源建模数据库进行3D结构预测以及密码子偏爱性分析。结果表明,E蛋白与其它黄病毒衣壳蛋白具有相似的功能,没有信号肽切割位点,在451-468和475-492aa区域含有跨膜区,为相对低表达基因,含有较多种类的稀有密码子,与人密码子使用频率较为接近。这为进一步研究DTMUV E基因的体外表达和宿主选择提供了分子生物学依据。To provide the reference for the selection of antigen domain and gene expression host system of duck Tembusu virus,one recombinant plasmid comprising an ORF about 1488 bp was sequenced from DTMUV QD genebank combing NCBI ORF Finder and BLAST analysis. By analysis of NCBI BLASTP and DNAstar evolutionary tree,the ORF encoded protein have high similarity with the E gene of Flaviviriade virus class. Then the conservative structure domain was determined by Conserved Domain tools,signal peptide was predicted using Signal P4.1 and the trans-membrane region was predicted by TMHMM 2.0. Applying Net NGlyc1.0 and Net Phos2.0,Prot Scale online,glycosylation sites,phosphorylation sites,hydrophobicity were analyzed. Finally,the fraction and using frequency of different codes in same amino acids were investigated by EMBOSS,and 3D structure was predicted by protein homology modeling analysis. Results showed：the E protein in this study have similar function with other Flaviviriade virus class capsid protein,no signal peptide cutting locus was discovered,and contained trans-membrane region in 451-468 and 475-492 AA area. And the E gene in this study which containing many kinds of rare codon,expression level was relatively low,codon usage frequency is close to with people. The study may provides the basis of molecular biology and contribute to in vitro expression selecting in further research.

关 键 词：鸭坦布苏病毒 生物学信息学 结构预测 

分 类 号：S852.65[农业科学—基础兽医学]