PRRSV山西株ORF5和Nsp2基因序列分析
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摘要
采用RT-PCR方法对2009-2011年山西省分离的5株猪繁殖与呼吸综合征病毒(Porcine reproductive and respiratory syndrome virus,PRRSV)的ORF5和Nsp2(2 503~3 269nt)基因进行克隆和测序,并对其基因序列和推导的氨基酸序列与国内外毒株进行了同源性分析。序列分析结果显示,5株分离株Nsp2基因与国内分离的PRRSV变异株(JXA1、HuN、HUN4、HUB1)的序列同源性最高,为96.8%~98.2%,且缺失位置一致,均存在2个位点30个氨基酸缺失;ORF5基因大小为603bp,编码200个氨基酸,第13、151位均为具有强毒特性的精氨酸(R),137位为丝氨酸(S),表明这5株均为野毒株,与国内分离的PRRSV变异株(JXA1、HuN、HUN4、HUB1)毒株的序列同源性最高,为96.5%~98.0%。结果表明,山西省内目前流行的PRRSV为Nsp2缺失30个氨基酸的变异毒株。
ORF5and Nsp2genes of 5PRRSV strains isolated from Shanxi province were amplified by RT-PCR,cloned and sequenced.The obtained sequences and the deduced amino acid were analyzed and compared with the other published PRRSV strains.Sequence comparision showed that Nsp2genes of the 5isolated strains were up to 96.8%-98.2%homology with JXA1,HuN,HUN4and HUB1isolated from China and had the same 30amino acid deletion.The ORF5genes were603bp in length and consist of 200amino acids and the site of 13and 151were Arginine(R)which were related to virulent strains,and the site of 137was serine(S)indicating that the 5isolated strains were wild strains.The nucleotide homology are 96.5%-98.0% comparing with JXA1,HuN,HUN4and HUB1isolated from China.The results showed the prevalent PRRSV strains in Shanxi were the Nsp2gene deleted 30amino acid,which laid the foundation for the scientific prevention and control of PRRS.
ORF5and Nsp2genes of 5PRRSV strains isolated from Shanxi province were amplified by RT-PCR,cloned and sequenced.The obtained sequences and the deduced amino acid were analyzed and compared with the other published PRRSV strains.Sequence comparision showed that Nsp2genes of the 5isolated strains were up to 96.8%-98.2%homology with JXA1,HuN,HUN4and HUB1isolated from China and had the same 30amino acid deletion.The ORF5genes were603bp in length and consist of 200amino acids and the site of 13and 151were Arginine(R)which were related to virulent strains,and the site of 137was serine(S)indicating that the 5isolated strains were wild strains.The nucleotide homology are 96.5%-98.0% comparing with JXA1,HuN,HUN4and HUB1isolated from China.The results showed the prevalent PRRSV strains in Shanxi were the Nsp2gene deleted 30amino acid,which laid the foundation for the scientific prevention and control of PRRS.
引文
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[11]刘飞,王开功,周碧君,等.贵州省PRRSV分离株Nsp2基因的克隆及序列分析[J].中国预防兽医学报,2010,32(12):924-928.
[2]董建国,刘永刚,石文达,等.高致病性PRRSV HuN株Nsp4的原核表达及其抗体水平检测[J].中国预防兽医学报,2011,33(3):219-222.
[3]Meulenberg J J M,Hulst M,De Meijer E J,et al.Lelystad virus,the causative agent of porcine epidemic abortion and respiratory syndrome(PEARS),is related to LDV and EAV[J].Virology,1993,192:62-72.
[4]安同庆,田志军,冷超良,等.我国大陆地区PRRSV的遗传衍变分析[A]//金宁一.第四次猪病学研讨会论文集[C].河南郑州:中国畜牧兽医学会动物传染病学分会,2010.
[5]范培虎,危艳武,郭龙军,等.2005—2010年我国部分地区PRRSV流行毒株的遗传变异分析[J].动物医学进展,2011,32(3):1-7.
[6]高志强,郭鑫,杨汉春,等.猪繁殖与呼吸综合征病毒缺失变异株的基因组特征[J].畜牧兽医学报,2005,36(6):578-584.
[7]Stadejek T,Stankevicius A,Storggaard T,et al.Identification of radically different variants of porcine reproductive and respiratory syndrome virus in Eastern Europe:towards a common ancestor for European and American viruses[J].J GenVirol,2002,83:1861-1873.
[8]周海范,夏平安,崔保安,等.猪繁殖与呼吸综合征病毒河南分离株ORF5基因的克隆与变异分析[J].中国兽医学报,2007,27(6):810-813.
[9]Wesley R D,Mengeling W L,Lager K M,et al.Differentiation of a porcine reproductive and respiratory syndrome virus vaccine strain from North American field strains by restriction fragment lengh polymorphism analysis of ORF5[J].J Vet Diagn Invest,1998,10:140-144.
[10]Ansari I H,Kwon B,Osorio F A,et al.Influenceof Nlinked glycosylation of porcine reproductive and respiratorysyndrome virus GP5on virus infectivity,antigenicity,and ability to induce neutralizing antibodies[J].J Virol,2006,80(8):3994-4004.
[11]刘飞,王开功,周碧君,等.贵州省PRRSV分离株Nsp2基因的克隆及序列分析[J].中国预防兽医学报,2010,32(12):924-928.