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PRRSV分离株Nsp2基因和ORF5基因的遗传变异分析
引用本文:王剑非,孙建全,张金强,迟小伟,刘杰,丛晓燕,时建立,李俊,吴家强.PRRSV分离株Nsp2基因和ORF5基因的遗传变异分析[J].家畜生态,2011(1):48-53.
作者姓名:王剑非  孙建全  张金强  迟小伟  刘杰  丛晓燕  时建立  李俊  吴家强
作者单位:山东省农业科学院;山东省畜禽疫病防治与繁育重点实验室;山东中医药大学附属医院;山东大学附属济南市中心医院;
基金项目:国家转基因生物新品种培育科技重大专项(2009ZX08009-143B); 山东省自然科学基金重点项目(Z2007D06); 山东省中青年科学家科研奖励基金(2009BSC02016); 山东省留学回国人员科技活动择优资助项目(鲁人社函[2009]341号)
摘    要:为了解山东省PRRSV流行毒株的遗传变异情况和分子流行病学背景,对2007~2009年分离到的14株山东省内的PRRSV流行毒株进行了Nsp2和ORF5基因序列的测定和分析。结果显示,与参考毒株JXA1相比,14株PRRSV Nsp2与ORF5基因的核苷酸同源性分别达到96.1%~99.8%和98.0%~99.8%,氨基酸序列同源性分别达到92.9%~100%和97.5%~99.5%。序列分析结果表明,14株毒株在Nsp2蛋白内部均存在编码30个氨基酸的碱基对的不连续缺失;ORF5基因编码的GP5蛋白不存在缺失,但存在点突变。遗传进化分析表明,07年分离毒株JQ与参考毒株JXA1亲缘关系很近;09年分离的12株PRRSV,2株仍与国内2006~2007年间的分离株在同一分支,9株已经处在不同分支,1株单独处在一个分支。2007~2009年山东地区流行的PRRSV分离株具有年度特征,无明显的地域特征。

关 键 词:猪繁殖与呼吸综合征病毒  分离  Nsp2基因  ORF5基因  变异分析

Genetic Variations of Nsp2 Gene and ORF5 Gene of Porcine Reproductive and Respiratory Syndrome Virus Isolates
WANGg Jian-fei,SUN Jian-quan,ZHANG Jin-qiang,CHI Xiao-wei,LIU Jie,CONG Xiao-yan,SHI Jianli,LI Jun,WU Jia-qiang.Genetic Variations of Nsp2 Gene and ORF5 Gene of Porcine Reproductive and Respiratory Syndrome Virus Isolates[J].Ecology of Domestic Animal,2011(1):48-53.
Authors:WANGg Jian-fei  SUN Jian-quan  ZHANG Jin-qiang  CHI Xiao-wei  LIU Jie  CONG Xiao-yan  SHI Jianli  LI Jun  WU Jia-qiang
Institution:WANGg Jian-fei1,SUN Jian-quan1,2,ZHANG Jin-qiang1,CHI Xiao-wei3,LIU Jie4,CONG Xiao-yan1,SHI Jianli1,LI Jun1,WU Jia-qiang1,2(1.Shandong Academy of Agricultural Sciences,Jinan,Shandong 250100,China,2.Shandong Provincial Key Lab of Animal Diseases Control and Breeding,3.Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Shandong 250012,4.Jinan Central Hospital Affiliated to Shandong University,Shandong 250013,China)
Abstract:To fully understand the knowledge of highly pathogenic porcine reproductive and respiratory syndrome(PRRSV) molecular epidemiology and genetic variation in Shandong area,a total of 14 PRRSV isolates were isolated in 2007~2009,and Nsp2 gene and ORF5 gene of these isolates were amplified by RT-PCR and sequenced.Comparing with JXA1 strain,Nsp2 gene shared 96.1%~99.8% of nucleotide homologies and 92.9%~100% of the deduced amino acid homologies,while the ORF5 gene shared 98.0%~99.8% of nucleotide homologies and 97.5%~99.5% of the deduced amino acid homologies.Sequence results showed that all the 14 PRRSV isolates had discontinuous 30 amino acids deletions in Nsp2;the GP5 protein encoded by ORF5 gene had a few mutation but no deletion.The phylogenetic tree revealed JQ strain isolated in 2007 had closer relationship with JXA1 strain.Among the 12 PRRSV strains isolated in 2009,two strains still locate in the same branch with the domestic strains isolated between 2006 and 2007,nine strains constitute an independent branch,and 1 strain(DLS-1) forms in a separate branch,indicating that those PRRSV isolates have no obvious geographical characteristics but have an annual feature in Shandong area.
Keywords:PRRSV  isolation  NSP2 gene  ORF5 gene  variation analysis  
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