猪繁殖与呼吸综合征病毒Nsp10基因的克隆及序列分析  

Cloning and sequence analysis of Nsp10 gene of PRRSV strain

作　　者：周思旋[1] 周碧君[1,2] 隆华 朱时杰[1] 文明[1,2] 王开功[1,2] 

机构地区：[1]贵州大学动物科学学院,贵州贵阳550025 [2]贵州大学动物疫病研究所,贵州贵阳550025 [3]贵州省畜牧兽医局,贵州贵阳550001

出　　处：《黑龙江畜牧兽医》2009年第10期10-13,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金：贵州省"十一五"农业科技攻关项目(黔科合NZ字[2005]3002);贵州省农业厅兽医科技计划项目(200806);贵阳市农业局科技计划项目([2008]动防-03号);贵州大学研究生创新基金项目(校研农200902)

摘　　要：根据猪繁殖与呼吸综合征病毒BJ-4序列,选择Nsp10基因保守区域设计并合成特异性引物,用RT-PCR方法扩增出猪繁殖与呼吸综合征病毒贵州地方株(命名为JL株)Nsp10基因片段,克隆到pMD18-T载体并测序,应用DNASTAR、C lustal_1.81和Mega序列分析软件进行同源性比较。结果表明:贵州JL株Nsp10基因的cDNA长699 bp,可编码233个氨基酸;贵州JL株Nsp10基因与早期分离毒株MLV株、BJ-4株、VR-2332株、CH-1a株、CH2002株和GS2004株相应序列的核苷酸同源性为91.7%～94.0%,氨基酸同源性为97.4%～98.7%;而与最近分离毒株HN2007株、SX2007株、GD2007株、YN2008株、GS2008株、XL2008株、TJ株和JXA1株的核苷酸同源性为98.3%～98.6%,氨基酸同源性为99.6%～100%。A pair of specific primers of Nsp10 gene was designed according to the gene sequence of PRRSV BJ -4 of American standard strain. The specific fragment of Guizhou isolate was been successfully amplified by RT - PCR and sequenced after being cloned directly into the pMD 18 -T. The homology of Nsp10 nucleotide and deduced amino acid were compared with different strains which shown on GenBank by the softwares of DNASTAR,Clustal 1.81 and Mega. It was showed that the cDNA of NsplO was 699 bp and it shared 91.7% -94.0% of nucleotide homology and 97.4 - 98.7 % of amino acid homo-logy with that of pristine stains （ MLV, BJ - 4, VR - 2332, CH - 1 a, CH2002 and GS2004 ） , respectively. And it showed 98.3% - 98.6% of nucleotide identity and 99.6% - 100% of amino acid identity with that of recent strainsc （ HN2007, SX2007, YN2008, GD2007, TJ and JXA1 ）, respectively.

关 键 词：猪繁殖与呼吸综合征病毒 Nsp10基因 克隆 序列分析 

分 类 号：S852.659.6[农业科学—基础兽医学]