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.
作者
张彩勤
王旭荣
杨增岐
赵炳武
宋长绪
ZHANG Cai-qin;WANG Xu-rong;YANG Zeng-qi;ZHAO Bing-wu;SONG Chang-xu(Northwest A&F Universty,Yangling Shaanxi712100,China;Institute of Veterinary Medicine,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,China)
出处
《西北农业学报》
CSCD
北大核心
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)
