| 1株基因重组猪繁殖与呼吸综合征病毒的新型变异和遗传进化分析 |
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| 引用本文： | 胡栋,朱迎春,赵情,庞恒,李传刚,王亭亭,常维山,彭军,吴家强.1株基因重组猪繁殖与呼吸综合征病毒的新型变异和遗传进化分析[J].中国兽医学报,2021(2):213-223. |
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| 作者姓名： | 胡栋 朱迎春 赵情 庞恒 李传刚 王亭亭 常维山 彭军 吴家强 |
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| 作者单位： | ;1.山东农业大学动物科技学院山东省动物生物工程与疾病防治重点实验室/农业部动物疫病病原生物学华东科学观测实验站;2.山东省农业科学院畜牧兽医研究所山东省畜禽疫病防治与繁育重点实验室 |
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| 基金项目： | 国家自然科学基金资助项目(31672587);山东省重点研发计划资助项目(2016GNC110013);山东省农业重大应用技术创新资助项目(SD2019XM007);山东省“双一流”建设资助项目(SYL2017YSTD11)。 |
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| 摘 要： | 对山东省某地区初诊为猪繁殖与呼吸综合征(porcine reproduction and respiatory syndrome,PRRS)的猪病料采用RT-PCR进行鉴定,获得1株PRRSV并命名为SD18。对该分离株的第2代细胞培养毒株进行全基因组测序,与其他参考毒株比较同源性并构建遗传进化树。结果显示,SD18毒株属于美洲型高致病性PRRSV变异株,该毒株Nsp2基因具有31个不连续的氨基酸缺失,已明显区别于以JXA1、HUN4、TJ株等为代表的30个不连续缺失高致病性分离株,其中第830位氨基酸位点是最新发现的缺失氨基酸。同时,该毒株的ORF3基因第17位插入1个丝氨酸(S),而ORF5基因编码的第13位和第151位氨基酸均为具有强毒特性的精氨酸(R),第137位则插入了具有野毒特性的丝氨酸(S),这4个氨基酸位点也表现出不同于以往高致病性毒株变异的特征。采用基因重组分析软件RDP4分析表明,该新型缺失株的多个部位发生了基因重组;SD18毒株病毒以美洲经典毒株VR2332作为重组的主要亲本毒株,疫苗株JXA1-R和新型毒株NADC30-like提供重组片段,多数重组变化集中在Nsp2蛋白区域,在非结构蛋白和次要蛋白区域也可见部分重组变化。本研究为深入探索PRRSV的遗传变异规律及相关生物学特性研究积累了数据。综合分析表明,山东地区SD18分离株Nsp2、GP3、GP5相应氨基酸均出现了较大变异,特别是Nsp2出现一个新的氨基酸缺失,可能是该毒株形成了一个新的独立变异分支的重要原因;这些变异也提示该分离毒株在病毒编码蛋白及其免疫原性等方面出现了较大变化。该病毒株的成功分离鉴定与遗传进化分析为PRRSV衍化及流行病学调查积累了最新数据,也为预防控制该病提供参考。
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| 关 键 词： | 猪繁殖与呼吸综合征病毒 变异毒株 遗传衍化 NSP2基因 基因重组 |
The novel variation and genetic evolution analysis on a recombinant porcine reproduction and respiratory syndrome virus |
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| HU Dong,ZHU Yingchun,ZHAO Qing,PANG Heng,LI Chuangang,WANG Tingting,CHANG Weishan,PENG Jun,WU Jiaqiang.The novel variation and genetic evolution analysis on a recombinant porcine reproduction and respiratory syndrome virus[J].Chinese Journal of Veterinary Science,2021(2):213-223. |
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| Authors: | HU Dong ZHU Yingchun ZHAO Qing PANG Heng LI Chuangang WANG Tingting CHANG Weishan PENG Jun WU Jiaqiang |
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| Institution: | (Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention/Experiment Station of Ministry of Agriculture and Rural Affairs for Animal Disease Pathogen Biology in East China,College of Animal Science and Technology,Shandong Agricultural University,Tai'an,Shandong 271018,China;Shandong Provincial Academy of Agricultural Sciences,Shandong Provincial Animal Disease Control and Breeding,Institute of Animal Science and Veterinary Medicine,Ji'nan 250100,China) |
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| Abstract: | The purpose of this study is to understand the genetic variation of a PRRSV epidemic strain.A PRRSV strain was obtained from the pig samples which were newly diagnosed as porcine reproductive and respiratory syndrome(PRRS)in a certain area of Shandong Province,and it was named SD18.The complete genome of the virus strain was sequenced,following by the homologycomparison of the whole genes with other reference strains,and then a genetic phylogenetic tree was constructed.The results showed that the SD18 strain belongs to the American type of highly pathogenic PRRSV variant.The Nsp2 gene of this strain has 31 discrete amino acid deletions,which was obviously different from the highly pathogenic isolates with 30 discontinuous amino acid deletion represented by JXA1,HUN4 and TJ strains.In particular,the 830 th amino acid site was a newly discovered missing amino acid.At the same time,the serine(S)was inserted at position 17 of the ORF3 gene of the strain.The amino acids 13 and 151 encoded by the ORF5 gene were all arginine(R),and the serine which was a characteristic of wild-type strain was inserted at position 137 of ORF5 gene.These four amino acid loci also showed different variation characteristics from the previous highly pathogenic strains.The result of gene recombination analysis by the software RDP4 showed that gene recombination occurred in many parts of the SD18 strain.The SD18 strain used the highly pathogenic strain VR2332 as the main parental strain for the recombinant,and most of the recombination changes were concentrated in the Nsp2 protein region,and partial recombination changes were also observed in the non-structural and minor protein regions.Comprehensive analysis showed that the amino acid sequences corresponding to Nsp2,GP3,and GP5 in SD18 PRRSV isolate in Shandong all showed large variations.In particular,a new amino acid deletion in Nsp2 may be an important reason for this strain to form a new independent variation branch.These mutations also suggest that the isolated strain has undergone major changes in the virus-encoded proteins and their immunogenicity.The successful isolation,identification and phylogenetic analysis of the virus strain have accumulated the latest data for the virus evolution and epidemiological investigation of PRRS,and also provide a reference for the prevention and control of the disease. |
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| Keywords: | PRRSV variant strain genetic variation Nsp2 gene gene recombinant |
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