猪繁殖与呼吸综合征河南毒株Henan-1的分离及其NSP2、ORF3、ORF5的序列分析  

Isolation and Sequence Analysis of NSP2,ORF3 and ORF5 of PRRSV Henan-1 Strain from Henan Province

作　　者：张彩勤[1] 王旭荣[1] 杨增岐[1] 赵炳武[2] 宋长绪[2] 

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100 [2]广东省农业科学院兽医研究所,广州510640

出　　处：《西北农业学报》2008年第1期1-6,共6页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家食品安全重大专项(2001BA804A31);广东省农业科技攻关(2003B21404、20052490100和2006B20301013);广东省自然科学基金(04002083);广东省自然基金重点项目(06105311)

摘　　要：将2007年1月河南省某猪场送检的疑似高热病病料处理后接种于Marc-145细胞，出现细胞聚集、固缩、脱落等特异性细胞病变。根据GenBank中猪繁殖与呼吸综合征病毒（PRRSV）的基因序列设计并合成针对NSP2、ORF3和ORF5基因的引物，RT-PCR扩增目的基因并将其克隆入pMD18-T载体进行测序。并与国内外已发表的毒株的序列进行比较。结果表明：分离毒株Henan-1是美洲型PRRSV，而且NSP2基因发生30个AA的不连续缺失；Henan-1的NSP2、ORF3和ORF5基因与JXA1、JX0612、HZ-X、HB-1（sh）、CH-1a、HB-2（sh）、SP、ATCC VR-2332、Henan-HN1、Resp PRRS／Repro MLV、PA8、BJ-4等毒株等位基因的核苷酸同源性分别为83．0％～99．0%、88．3％～99．2％、88．6％～99．5％，推导的氨基酸同源性分别为66．7％～97．9％、84．7％～98．8％、87．1％～99．5％；而ORF3和ORF5基因与LV4．2．1等位基因核苷酸同源性分别为64．7％和64．2％，推导的氨基酸同源性分别为56．7％和61．0％。基因系统发育树分析表明：Henan-1与JXA1、JX0612株的亲缘关系很近，与HB-1（sh）亲缘关系较近，与HZ-X、CH-1a、HB-2（sh）、SP、RespPRRS／Repro MLV等毒株处于不同的分支。The samples collected from Henan province were inoculated to Marc-145 cells, the PRRSV specific cytopathogenic effect （CPE） was observed and the isolates named Henan-1 obtained. NSP2, ORF3 and ORF5 genes of PRRSV Henan-1 strain were amplified by RT-PCR and sequenced after being cloned directly into the pMD18-T. The sequences of NSP2, ORF3 and ORF5 genes of Henan-1 strain were compared with sequences of other PRRSV strains in GenBank by Blast and DNAStar software. The results demonstrated that Henan-1 strain could classified into the North American type of PRRSV and its NSP2 had 30 amino acids deletion . The NSP2, ORF3 and ORF5 of Henan-1 shared 83.0%-99.0%,88. 3%-99. 2% and 88. 6%-99.5%of nucleotide homology , and 66. 7%-97.9%,84.7%-98.8% and 87.1%-99.5% of amino acid identity , with that of JXA1,JX0612,HZX, HB-1 （sh） ,CH-1a ,HB-2（sh）, SP, ATCC VR-2332 ,Henan-HN1,PAS,BJ-4 and Resp PRRS/Repro MLV respectively. The ORF3 and ORF5 of Henan-1 shared only 64.7%, 64.2% of nucleotide homology, and 56. 7%, 61.0% of amino acid homology with that of LV4. 2. 1 respectively. The phylogenetic tree revealed Henan 1 had closer relationship with JXA1 and JX0612 strains than that of HB-1（sh） strain.

关 键 词：猪繁殖与呼吸综合征病毒 Henan-1毒株 序列分析 

分 类 号：S852.65[农业科学—基础兽医学]