伪狂犬病病毒FJ-2015株主要免疫原基因的遗传进化分析
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摘要
为了解伪狂犬病病毒(PRV)变异株主要免疫原相关基因的序列遗传特征,本研究对PRV FJ-2015株g B、g C和g D基因进行克隆和测序分析。结果显示,FJ-2015株3个主要免疫原基因推导氨基酸序列与2012年以来国内新分离PRV变异株同源性较高,分别为99.7%~100%、99.6%~100%和98.5%~99.8%;而与国外分离株氨基酸同源性较低,分别为96.9%~97.5%、93.1%~93.3%和97.3%~98.8%,其中与疫苗株Bartha-K61同源性最低,为96.9%、93.1%和97.3%;氨基酸比对显示,与经典病毒株相比,该病毒株发生多个位点的一致性替换、插入或缺失,并处于重要的抗原表位区;遗传进化关系表明,该病毒株g B基因遗传演化与分离时间和分离国家相关,g C基因显示遗传多样性,而g D基因具有一定的福建区域性遗传演化关系。以上结果表明,PRV FJ-2015属于PRV变异株,与其它分离株存在一定的遗传差异。本研究丰富了猪PRV流行的分子特征。
To understand the genetic characteristics of the main immunogen related genes of variant pseudorabies virus(PRV),the g B, g C and g D genes of the PRV FJ-2015 strain were cloned and sequenced in this study. The results demonstrated that the amino acid sequences of the 3 main immunogen genes of the PRV FJ-2015 strain shared high homologies with that of variant PRV strains isolated since 2012, and the homologies were 99.7%-100%, 99.6%-100% and 98.5%-99.8%, respectively. However, the g B, g C and g D genes of the PRV FJ-2015 strain shared lower homologies with that of the foreign PRV strains, the homologies were 96.9%-97.5%, 93.1%-93.3% and 97.3%-98.8%, respectively, and the lowest homologies were shared with Bartha-K61 vaccine strain, which were 96.9%, 93.1% and 97.3%, respectively. The amino acid comparison showed that the multiple variations took place on the PRV FJ-2015 strain compared with the classical PRV strains, including substitutions, insertions and/or deletions,some of which were located in epitopes. Phylogenetic analysis based on these gene sequences indicated that the g B gene differed significantly depending upon the isolation countries and isolated times, the g C gene showed genetic diversity and the g D gene was unique in the Fujian region. The above results indicated that PRV FJ-2015 strain was a variant pseudorabies virus which had certain genetic differences from other PRV isolate strains. This study enrichs the molecular epidemiology of PRV.
To understand the genetic characteristics of the main immunogen related genes of variant pseudorabies virus(PRV),the g B, g C and g D genes of the PRV FJ-2015 strain were cloned and sequenced in this study. The results demonstrated that the amino acid sequences of the 3 main immunogen genes of the PRV FJ-2015 strain shared high homologies with that of variant PRV strains isolated since 2012, and the homologies were 99.7%-100%, 99.6%-100% and 98.5%-99.8%, respectively. However, the g B, g C and g D genes of the PRV FJ-2015 strain shared lower homologies with that of the foreign PRV strains, the homologies were 96.9%-97.5%, 93.1%-93.3% and 97.3%-98.8%, respectively, and the lowest homologies were shared with Bartha-K61 vaccine strain, which were 96.9%, 93.1% and 97.3%, respectively. The amino acid comparison showed that the multiple variations took place on the PRV FJ-2015 strain compared with the classical PRV strains, including substitutions, insertions and/or deletions,some of which were located in epitopes. Phylogenetic analysis based on these gene sequences indicated that the g B gene differed significantly depending upon the isolation countries and isolated times, the g C gene showed genetic diversity and the g D gene was unique in the Fujian region. The above results indicated that PRV FJ-2015 strain was a variant pseudorabies virus which had certain genetic differences from other PRV isolate strains. This study enrichs the molecular epidemiology of PRV.
引文
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[8]夏德利,黄立平,危艳武,等.猪伪狂犬病病毒流行毒株gC基因的分子特征及其对小鼠的毒力试验[J].中国兽医科学,2017,47(6):713-720.
[9]Wang Yin-biao,Qiao Song-lin,Li Xue-wu,et al.Molecular epidemiology of outbreak-associated pseudorabies virus(PRV)strains in central China[J].Virus Genes,2015,50(3):401-409.
[10]杨文萍,顾真庆,孙海凤,等.伪狂犬病毒流行毒株的抗原性和血清中和特性分析[J].畜牧与兽医,2014,46(10):11-14.
[11]Zaripov M M,Morenkov O S,Shmatchenko V V,et al.Immunological and functional characteristics of epitopes and regions of gB glycoprotein of Aujeszky's disease virus[J].Vopr Virusol,2011,46(2):41-45.
[12]刘存,陈书民,刘砚涵,等.伪狂犬病病毒Qihe547分离株主要保护性抗原基因序列的变异分析[J].中国预防兽医学报,2016,38(4):331-334.
[13]范克伟,戴爱玲,李晓华,等.闽西地区猪伪狂犬病病毒变异株gC基因克隆与序列分析[J].中国兽医杂志,2016,52(1):47-48.
[14]曾显成,吴异健,陈家祥,等.猪伪狂犬病病毒FZ株的分离鉴定及gD基因序列分析[J].福建农林大学学报(自然版),2011,40(5):501-510.
[2]Schroter C,Vallbracht M,Altenschmidt J,et al.Mutations in pseudorabies virus glycoproteins gB,gD,and gH functionally compensate for the absence of gL[J].J Virol,2015,90(5):2264-2272.
[3]Bohm S W,Eckroth E,Backovic M,et al.Structure-based func-tional analyses of domains II and III of pseudorabies virus glycoprotein H[J].J Virol,2015,89(2):1364-1376.
[4]Mulder W,Pol J,Kimman T,et al.Glycoprotein D-negative pseudorabies virus can spread transneuronally via direct neuron-to-neuron transmission in its natural host,the pig,but not after additional inactivation of gE or gI[J].J Virol,1996,70(4):2191-2200.
[5]Klupp B G,Lomniczi B,Visser N,et al.Viral pathogens and immunity mutations affecting the UL21 gene contribute to avirulence of pseudorabies virus vaccine strain Bartha[J].Virology,1995,212(2):466-473.
[6]童光志,陈焕春.伪狂犬病流行现状及我国应采取的防制措施[J].中国兽医学报,1999,19(1):1-2.
[7]彭金美,安同庆,赵鸿远,等.猪伪狂犬病病毒新流行株的分离鉴定及抗原差异性分析[J].中国预防兽医学报,2013,35(1):1-4.
[8]夏德利,黄立平,危艳武,等.猪伪狂犬病病毒流行毒株gC基因的分子特征及其对小鼠的毒力试验[J].中国兽医科学,2017,47(6):713-720.
[9]Wang Yin-biao,Qiao Song-lin,Li Xue-wu,et al.Molecular epidemiology of outbreak-associated pseudorabies virus(PRV)strains in central China[J].Virus Genes,2015,50(3):401-409.
[10]杨文萍,顾真庆,孙海凤,等.伪狂犬病毒流行毒株的抗原性和血清中和特性分析[J].畜牧与兽医,2014,46(10):11-14.
[11]Zaripov M M,Morenkov O S,Shmatchenko V V,et al.Immunological and functional characteristics of epitopes and regions of gB glycoprotein of Aujeszky's disease virus[J].Vopr Virusol,2011,46(2):41-45.
[12]刘存,陈书民,刘砚涵,等.伪狂犬病病毒Qihe547分离株主要保护性抗原基因序列的变异分析[J].中国预防兽医学报,2016,38(4):331-334.
[13]范克伟,戴爱玲,李晓华,等.闽西地区猪伪狂犬病病毒变异株gC基因克隆与序列分析[J].中国兽医杂志,2016,52(1):47-48.
[14]曾显成,吴异健,陈家祥,等.猪伪狂犬病病毒FZ株的分离鉴定及gD基因序列分析[J].福建农林大学学报(自然版),2011,40(5):501-510.