1株猪圆环病毒3型全基因组克隆与序列分析
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摘要
【目的】克隆获得1株猪圆环病毒3型(PCV3)全基因组序列并进行序列分析。【方法】以PCV3阳性临床样品为模板,用PCR技术扩增PCV3全基因组片段,随后进行克隆、序列测定与分析。【结果】扩增出1株PCV3全基因组片段,序列测定结果显示,这株PCV3全基因组大小为2 000bp,与其他国内外14株PCV3的核苷酸同源性均达98.6%以上,表明获得1株PCV3全基因组序列,且不同PCV3之间的全基因组核苷酸有非常高的同源性。PCV3与PCV1、PCV2之间的全基因组核苷酸同源性都低于50%。系统发育显示,3种PCV分别处于明显不同的分支,表明各自属于不同的基因型。【结论】获得1株PCV3全基因组序列,其与其他PCV3毒株之间的核苷酸同源性很高。
【Objective】The aim of this study is to clone and analyze the complete genome sequence of a porcine circovirus type 3(PCV3)strain.【Methods】Clinical PCV3 samples were used as templates to amplify the complete genome fragment of PCV3 with PCR method,and then the fragment was cloned,sequenced and analyzed.【Results】Sequence determination results showed that the complete genome of this PCV3 strain was 2 000 bp in size and shared more than 98.6%of nucleotide homology with other 14 PCV3 strains at home and abroad,indicating that the complete genome sequence of a PCV3 strain was obtained and the nucleotide homology of the complete genome of different PCV3 strains was very high.The nucleotide homology of the complete genome between PCV3 and PCV1 or PCV2 was all less than 50%.The phylogenetic analysis showed that the 3 kind of PCVs were in distinct branches and revealed that they belonged to different genotypes.【Conclusion】The complete genome sequence of a PCV3 strain was obtained and shared very high nucleotide homology with other PCV3 strains.
【Objective】The aim of this study is to clone and analyze the complete genome sequence of a porcine circovirus type 3(PCV3)strain.【Methods】Clinical PCV3 samples were used as templates to amplify the complete genome fragment of PCV3 with PCR method,and then the fragment was cloned,sequenced and analyzed.【Results】Sequence determination results showed that the complete genome of this PCV3 strain was 2 000 bp in size and shared more than 98.6%of nucleotide homology with other 14 PCV3 strains at home and abroad,indicating that the complete genome sequence of a PCV3 strain was obtained and the nucleotide homology of the complete genome of different PCV3 strains was very high.The nucleotide homology of the complete genome between PCV3 and PCV1 or PCV2 was all less than 50%.The phylogenetic analysis showed that the 3 kind of PCVs were in distinct branches and revealed that they belonged to different genotypes.【Conclusion】The complete genome sequence of a PCV3 strain was obtained and shared very high nucleotide homology with other PCV3 strains.
引文
[1]Phan T G,Giannitti F,Rossow S,Marthaler D,Knutson T,Li L,Deng X,Resende T,Vannucci F and Delwart E.Detection of a novel circovirus PCV3in pigs with cardiac and multi-systemic inflammation[J].Virology Journal,2016,13(1):184
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[10]杨红杰,于长青,林树伯,于红欣,周双海,李继良.伪狂犬病病毒gB基因荧光定量PCR检测方法的建立[J].北京农学院学报,2015,30(3):48-51
[11]Tischer I,Mields W,Wolff D,Vagt M,Griem W.Studies on epidemiology and pathogenicity of porcine circovirus[J].Archives of Virology,1986,91(3-4):271-276
[12]Allan G M,McNeilly F,Cassidy J P,Reilly G A,Adair B,Ellis W A,McNulty M S.Pathogenesis of porcine circovirus:experimental infection of colostrums deprived piglets and examination of pig foetal material[J].Veterinary Microbiology,1995,44(1):49-64
[13]Opriessnig T,Meng X J,Halbur P G.Porcine circoviruses type 2-associated disease:update on current terminology,clinical manifestations,pathogenesis,diagnosis,and intervention strategies[J].Journal of Veterinary Diagnostic Investigation,2007,19:591-615
[14]刘建奎,魏春华,陈小燕,虞慧芸,刘佳悦,范佳霖,戴爱玲,吴昌标,杨小燕.福建省新发猪圆环病毒3型流行病学调查及遗传变异分析[J].中国预防兽医学报,2018(2):160-163
[15]庞笑语,王建昌,韩庆安,王金凤,张若曦,李睿文,董世山,孙继国,袁万哲.河北省猪群中检出猪圆环病毒3型[J].中国动物检疫,2017,34(11):70-73
[16]Hamel A L,Lin L L,Nayar G P.Nucleotide sequence of porcine circovirus associated with postweaning multisystemic wasting syndrome in pigs[J].Journal of Virology,1998,72(6):5262-5267
[2]Palinski R,Pi1eyro P,Shang P,Yuan F,Guo R,Fang Y,Byers,E Hause B M.A novel porcine circovirus distantly related to known circoviruses is associated with porcine dermatitis and nephropathy syndrome and reproductive failure[J].Journal of Virology,2016,91(1):1-16
[3]Stadejek T,Wozniak A,Milek D,Biernacka K.First detection of porcine circovirus type 3on commercial pig farms in Poland[J].Transboundary&Emerging Diseases,2017,64(5):1-4
[4]Kwon T,Yoo S J,Park C K,Lyoo Y S.Prevalence of novel porcine circovirus 3in Korean pig populations[J].Veterinary Microbiology,2017,207:178-180
[5]Ku X,Chen F,Li P,Wang Y,Yu X,Fan S,Qian P,Wu M,He Q.Identification and genetic characterization of porcine circovirus type 3in China[J].Transboundary&E-merging Diseases,2017,64(3):703
[6]Zheng S,Wu X,Zhang L,Xin C,Liu Y,Shi J,Peng Z,Xu S,Fu F,Yu J,Sun W,Xu S,Li J,Wang J.The occurrence of porcine circovirus 3without clinical infection signs in Shandong Province[J].Transboundary&Emerging Diseases,2017,64(5):1337-1341
[7]Fan S,Ku X,Chen F,Wang Y,Yu Y,He Q.Complete genome sequence of a novel porcine circovirus type 3strain,PCV3/CN/Hubei-618/2016,isolated from China[J].Genome Announcements,2017,5(15):1-17
[8]Mankertz A,Mankertz J,Wolf K,Buhk H J.Identification of a protein essential for replication of porcine circovirus[J].Journal of General Virology,1998,79(2):381-384
[9]Nawagitgul P,Morozov I,Bolin S R,Harms P A,Sorden SD,Paul P S.Open reading frame 2of porcine circovirus type2encodes a major capsid protein[J].Journal of General Virology,2000,81(9):2281-2287
[10]杨红杰,于长青,林树伯,于红欣,周双海,李继良.伪狂犬病病毒gB基因荧光定量PCR检测方法的建立[J].北京农学院学报,2015,30(3):48-51
[11]Tischer I,Mields W,Wolff D,Vagt M,Griem W.Studies on epidemiology and pathogenicity of porcine circovirus[J].Archives of Virology,1986,91(3-4):271-276
[12]Allan G M,McNeilly F,Cassidy J P,Reilly G A,Adair B,Ellis W A,McNulty M S.Pathogenesis of porcine circovirus:experimental infection of colostrums deprived piglets and examination of pig foetal material[J].Veterinary Microbiology,1995,44(1):49-64
[13]Opriessnig T,Meng X J,Halbur P G.Porcine circoviruses type 2-associated disease:update on current terminology,clinical manifestations,pathogenesis,diagnosis,and intervention strategies[J].Journal of Veterinary Diagnostic Investigation,2007,19:591-615
[14]刘建奎,魏春华,陈小燕,虞慧芸,刘佳悦,范佳霖,戴爱玲,吴昌标,杨小燕.福建省新发猪圆环病毒3型流行病学调查及遗传变异分析[J].中国预防兽医学报,2018(2):160-163
[15]庞笑语,王建昌,韩庆安,王金凤,张若曦,李睿文,董世山,孙继国,袁万哲.河北省猪群中检出猪圆环病毒3型[J].中国动物检疫,2017,34(11):70-73
[16]Hamel A L,Lin L L,Nayar G P.Nucleotide sequence of porcine circovirus associated with postweaning multisystemic wasting syndrome in pigs[J].Journal of Virology,1998,72(6):5262-5267