猪繁殖与呼吸综合征病毒强弱毒株全基因序列比对分析
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摘要
为了解猪繁殖与呼吸综合病毒(PRRSV)致弱的分子学基础,对PRRSV JXA1株第5代(强毒)与第86代毒株(致弱毒)进行全基因组测序,将JXA1原始毒株和JXA1致弱毒株与其他2对PRRSV原始毒株/弱毒疫苗株进行基因序列比对分析。结果显示,这3对毒株(1对JXA1原始毒株/致弱毒株、2对PRRSV原始毒株/弱毒疫苗株)中结构蛋白的氨基酸突变率较高,表明毒力的改变与结构蛋白的改变关系更大;将JXA1株与JXA1-86株序列比对分析,发现编码Nsp5、Nsp6、Nsp8、Nsp12、M、N蛋白的氨基酸未发生变化,表明JXA1株毒力的降低可能与这些蛋白无关;将各代次PRRSV序列比对分析,发现在第70代到第86代序列之间,ORF1a中A_(928)V、I_(1155)M、E_(1629)D,GP2中I_(118)V,GP3中H_(79)N,GP4中I_(124)V,GP5中K_(59)N发生了改变,表明其毒力的减弱可能与以上几处变异有关。
To study the attenuation molecular basis of porcine reproductive and respiratory syndrome virus(PRRSV),whole genomes of PRRSV JXA1-5(parental strain)and JXA1-86(attenuated strain)were sequenced,compared and analyzed.And genomic sequences of JXA1/JXA1-86 were compared with the other two pairs of virulent parental/attenuated vaccine strains.The results showed that there was higher amino acid mutation rate in structural proteins among the three pairs of strains indicating that PRRSV JXA1 strain virulence changes were more related to structure proteins variations.Compared with JXA1,the amino acids encoding Nsp5,Nsp6,Nsp8,Nsp12,M and N proteins of JXA1-86 strain did not change,suggesting that virulence attenuating might not be relevant to those proteins.The genomic sequences of each generation PRRSV sequence were compared and analyzed,and the results showed that the variations among A_(928)V,I_(1155)M,E_(1629)D in ORF1 a,I_(118)V in GP2,H_(79)N in GP3,I_(124)V in GP4 and K_(59)N in GP5 between 70th and 86 th generations were found,which turned out that virulence attenuating might be associate with those variations.
To study the attenuation molecular basis of porcine reproductive and respiratory syndrome virus(PRRSV),whole genomes of PRRSV JXA1-5(parental strain)and JXA1-86(attenuated strain)were sequenced,compared and analyzed.And genomic sequences of JXA1/JXA1-86 were compared with the other two pairs of virulent parental/attenuated vaccine strains.The results showed that there was higher amino acid mutation rate in structural proteins among the three pairs of strains indicating that PRRSV JXA1 strain virulence changes were more related to structure proteins variations.Compared with JXA1,the amino acids encoding Nsp5,Nsp6,Nsp8,Nsp12,M and N proteins of JXA1-86 strain did not change,suggesting that virulence attenuating might not be relevant to those proteins.The genomic sequences of each generation PRRSV sequence were compared and analyzed,and the results showed that the variations among A_(928)V,I_(1155)M,E_(1629)D in ORF1 a,I_(118)V in GP2,H_(79)N in GP3,I_(124)V in GP4 and K_(59)N in GP5 between 70th and 86 th generations were found,which turned out that virulence attenuating might be associate with those variations.
引文
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[16]Yan Y L,Guo X,Ge X N,et al.Monoclonal antibody and porcine antisera recognized B-cell epitopes of Nsp2protein of a Chinese strain of porcine reproductive and respiratory syndrome virus[J].Virus Res,2007,126(1-2):207-215.
[17]李志杰,丁壮,孟轲音,等.PRRSV缺失变异毒株JL/07/SW株的分离鉴定及序列分析[J].中国兽医学报,2009,29(7):830-835.
[18]Fang Y,Snijder E J.The PRRSV replicase:Exploring the multifunctionality of an intriguing set of nonstructural proteins[J].Virus Res,2010,154(1-2):61-76.
[19]Music N,Gagono C A.The role of porcine reproductive and respiratory syndrome(PRRS)virus structural and non-structural proteins in virus pathogenesis[J].Anim Health Res Rev,2010,11(2):135-163.
[20]Brown E,Lawson S,Welbon C,et al.Antibody response to porcine reproductive and respiratory syndrome virus(PRRSV)nonstructural proteins and implications for diagnostic detection and differentiation of PRRSV typesⅠandⅡ[J].Vaccine Immunol,2009,16(5):628-635.
[2]Changhoon P,Hwi W S,Ikjae K,et al.A new modified live porcine reproductive and respiratory syndrome vaccine improves growth performance in pigs under field conditions[J].Clinical and Vaccine Immunology,2014,21(9):1350-1356.
[3]Kim J M,Kwon W Y,Choi E J,et al,Complete genome sequence of type 1porcine reproductive and respiratory syndrome virus strain E38,isolated from south Korea with a novel deletion[J].Genome Announcements,2015,3(5):1180-1194.
[4]李桃梅,赖志,崔立.猪繁殖与呼吸综合征病毒垂直传播及其危害研究进展[J].中国畜牧兽医,2015,42(5):1311-1317.
[5]郭宝清,陈章水,刘文兴,等.从疑似PRRS流产胎儿分离PRRSV的研究[J].中国畜禽传染病,1996,18(2):1-4.
[6]童光志,周艳君,都晓芳,等.高致病性PRRSV的分离鉴定及其分子流行病学分析[J].中国预防兽医学报,2007,29(5):321-326.
[7]Han W,Wu J J,Deng X Y,et al.Molecular mutations associated with the in vitro passage of virulent porcine reproductive and respiratory syndrome virus[J].Virus Genes,2009,38(2):276-284.
[8]Kimman T G,Cornelissen L A,Moormann R J,et al.Challenges for porcine reproductive and respiratory syndrome virus(PRRSV)vaccinology[J].Vaccine,2009,27(28):3704-3718.
[9]Zhou L,Yang H.Porcine reproductive and respiratory syndrome in China[J].Virus Res,2010,154:31-37.
[10]Yu X L,Zhou Z,Cao Z,et al.Assessment of the safety and efficacy of an attenuated live vaccine based on highly pathogenic porcine reproductive and respiratory syndrome virus[J].Clinical and Vaccine Immunology,2015,22(5):493-502.
[11]Leng X,Li Z,Xia M,et al.Mutations in the genome of the highly pathogenic porcine reproductive and respiratory syndrome virus potentially related to attenuation[J].Vet Microbiol,2012,157(1-2):50-60.
[12]Yu X L,Chen N H,Deng X Y,et al.Genomic sequencing reveals mutations potentially related to the over attenuation of a highly pathogenic porcine reproductive and respiratory syndrome virus[J].Clinical and Vaccine Immunology,2013,20(4):613-619.
[13]Zhou F,Zhao J,Chen L,et al.Complete genome sequence of a novel porcine reproductive and respiratory syndrome virus that emerged in China[J].Genome Announcements,2015,3(4):e0072-15.
[14]Shen S,Kwang J,Liu W,et al.Determination of the complete nucleotide sequence of a vaccine strain of porcine reproductive and respiratory syndrome virus and identification of the NSP2gene with a unique insertion[J].Arch Virol,2000,145(5):871-883.
[15]de Lima M,Pattnaik A K,Flores E F,et al.Serologic marker candidates identified amongst B-cell linear epitopes of Nsp2and structural proteins of a North American strain of porcine reproductive and respiratory syndrome virus[J].Virology,2006,353(2):410-421.
[16]Yan Y L,Guo X,Ge X N,et al.Monoclonal antibody and porcine antisera recognized B-cell epitopes of Nsp2protein of a Chinese strain of porcine reproductive and respiratory syndrome virus[J].Virus Res,2007,126(1-2):207-215.
[17]李志杰,丁壮,孟轲音,等.PRRSV缺失变异毒株JL/07/SW株的分离鉴定及序列分析[J].中国兽医学报,2009,29(7):830-835.
[18]Fang Y,Snijder E J.The PRRSV replicase:Exploring the multifunctionality of an intriguing set of nonstructural proteins[J].Virus Res,2010,154(1-2):61-76.
[19]Music N,Gagono C A.The role of porcine reproductive and respiratory syndrome(PRRS)virus structural and non-structural proteins in virus pathogenesis[J].Anim Health Res Rev,2010,11(2):135-163.
[20]Brown E,Lawson S,Welbon C,et al.Antibody response to porcine reproductive and respiratory syndrome virus(PRRSV)nonstructural proteins and implications for diagnostic detection and differentiation of PRRSV typesⅠandⅡ[J].Vaccine Immunol,2009,16(5):628-635.