珠三角地区鸭坦布苏病毒的全基因序列测定与分析  

Molecular cloning and sequence analyses of the complete genome of duck Tembusu virus isolated from the Pearl River Delta region

作　　者：张克山[1] 陈芳艳[2] 刘金[1] 蔡丝丝[1] 刘湘红[1] 张靖鹏[1] 陈瑞爱[1,3] 王林川[1,3] 

机构地区：[1]华南农业大学兽医学院,广东广州510642 [2]华南农业大学动物科学学院,广东广州510642 [3]广东大华农动物保健品股份有限公司,广东新兴527400

出　　处：《华南农业大学学报》2015年第3期13-19,共7页Journal of South China Agricultural University

基　　金：广东大华农动物保健品股份有限公司科研基金(113A2012206-1)

摘　　要：【目的】了解2010年以来分离于珠三角地区鸭坦布苏病毒病（Duck Tembusu virus，DTMUV）GDNS2010．1、GDNS2010．2、GDZQ2012、GDPY2013株的分子特征．【方法】参考GenBank收藏的DTMUVJS804株，共设计合成11对特异性引物，扩增了4株病毒的全基因序列片段．【结果和结论】4株病毒全长约为10990bp，无Ploy（A）尾结构，仅含有1个大的ORF，共编码3426个氨基酸，5′非编码区（5′UTR）各含有94hp．在3′非编码区（3′UTR），GDNS2010．1、GDNS2010．2毒株在103395～103411bp处缺失10个碱基．联合前期试验已测定的2株病毒序列（GDHZ2012．1、GDHZ2012．2），用DNAstar和MEGA6．0序列分析软件将6株病毒同GenBank收藏的国内的其他坦布苏病毒株比对，发现核酸序列相似性在98％以上，与Ntaya病毒和Sitiawan病毒相似性较大，相似性都在73％左右；与西尼罗河病毒、日本乙型脑炎病毒、黄热病毒等相似性皆低于63％．6株病毒囊膜蛋白E核酸序列相似性为97．5％～99．9％，推导氨基酸序列相似性在97％以上，其中同一地区分离的毒株相似性最高，达99．9％；在囊膜蛋白E154位存在一潜在的糖基化位点Asn-Try-Ser，在E蛋白结构域Ⅱ第E289位，极性带正电荷的Lys变为极性带负电荷的Glu，推测这一位点可能为病毒的毒力位点．【Objective】In order to comprehend the molecular characteristics of duck Tembusu virus,the isolations of GDNS2010. 1,GDNS2010. 2,GDZQ2012,GDPY2013 in the Pearl River Delta region were analyzed.【Method】From the reference strains( Gen Bank accession no. JS804),eleven pairs of specific primers were designed and synthesized to amplify genome sequence fragments of GDNS2010. 1,GDNS2010. 2,GDZQ2012,GDPY2013. The whole genome sequences of these four viruses were obtained. The complete genome sequences of four viruses were obtained by sequencing and splicing. 【Re-sult and conclusion】Analyses showed that the full length of these four viruses was 10 990 bp,no Ploy( A) tail structure comprising one large ORF,encoding 3 426 amino acids. The 5' non-coding region( 5'UTR) contained 94 bp. Ten bases were missing from 103 395 bp to 103 411 bp of GDNS2010. 1 and GDNS2010. 2 strains in the 3' non-coding region( 3' UTR). Two sequences of GDHZ2012. 1 and GDHZ2012. 2 preliminarily determined were joined for a sequence analysis. DNAstar and MEGA6. 0were used to analyze the sequences of six virus strains. Comparisons with other domestic Tembusu viruses recorded in Gen Bank showed that the similarity of the nucleic acid sequence was more than 98%. In comparison with Ntaya virus,Sitiawan virus,the similarity was about 73%. Compared with West Nile virus,Japanese encephalitis virus and yellow fever virus,the similarities were less than 63%. The similarity for nucleic acid sequence of envelope protein in the six isolates remained between 97. 5%- 99. 9%;the similarity of deduced amino acid sequence was found to be more than 97% whereas isolates from the same area had 99. 9% similarity. In the site of E154 in envelope,there are a potential glycosylation site Asn-Try-Ser and a potential virulence loci in E289.

关 键 词：鸭坦布苏病毒 克隆测序 全基因组序列 囊膜蛋白毒力位点 鸭黄病毒 

分 类 号：S855.3[农业科学—临床兽医学]