K亚群禽白血病病毒分离株全基因组测序及序列分析
|
摘要
K亚群禽白血病病毒(ALV-K)是近年来从地方品种鸡分离鉴定的新亚群ALV。本试验在对江苏某原种鸡场保存的琅琊鸡开展禽白血病净化过程中,分离并鉴定1株ALV-K,命名为JS13LY19。为探明其基因组来源及特征,对JS13LY19分离株前病毒DNA进行了分段克隆和测序,获得全长基因组序列,并与各亚群ALV参考株进行比对分析。结果显示,JS13LY19分离株符合复制完整型C型反转录病毒特征,缺乏肿瘤基因。其gp85基因相较于其他亚群ALV,与ALV-K参考株遗传进化关系最为接近,与ALV-K原型株JS11C1一致性最高(99.2%);而gag、pol、gp37、LTR、UTR及JS13LY19与内源性ALV显示出更高的一致性(92.0%～99.4%);其LTR U3区比大部分外源性ALV LTR少了1个CAAT enhancer盒、PRE盒、CArG盒及Y盒。JS13LY19分离株极有可能是JS11C1与内源性ALV重组产生,且具有内源性ALV LTR及U3区转录调控元件的部分缺失,可能使JS13LY19转录能力下降而致病性降低。
Avian leukosis virus subgroup K(ALV-K) is a new subgroup of ALV isolated and identified from indigenous chicken breeds in recent years.In this study,a strain named JS13 LY19 isolated from Langya chicken was identified as ALV-K.To find out the molecular characteristics and evolutionary relationships of JS13 LY19,the proviral genome was sequenced and analyzed with other reference strains of ALV.The JS13 LY19 isolate had a genetic organization typical of replication-competent C retroviruses lacking oncogenes.Sequence analysis showed that the gp85 gene of JS13 LY19 had the highest identity to that of JS11 C1,the prototype of ALV-K,but gag,pol,gp37,LTR and UTR genes had more identity to those of endogenous ALV.A CAAT enhancer box,a PRE box,a CArG box and a Y box in U3 of JS13 LY19 genome was found to be deleted compared with that of exogenous ALV.Therefore,it is speculated that JS13 LY19 is generated by the recombination of JS11 C1 and endogenous ALV.Having endogenous ALV LTR and partial deletions of transcription regulatory elements may reduce the transcription capacity and pathogenicity of JS13 LY19.
Avian leukosis virus subgroup K(ALV-K) is a new subgroup of ALV isolated and identified from indigenous chicken breeds in recent years.In this study,a strain named JS13 LY19 isolated from Langya chicken was identified as ALV-K.To find out the molecular characteristics and evolutionary relationships of JS13 LY19,the proviral genome was sequenced and analyzed with other reference strains of ALV.The JS13 LY19 isolate had a genetic organization typical of replication-competent C retroviruses lacking oncogenes.Sequence analysis showed that the gp85 gene of JS13 LY19 had the highest identity to that of JS11 C1,the prototype of ALV-K,but gag,pol,gp37,LTR and UTR genes had more identity to those of endogenous ALV.A CAAT enhancer box,a PRE box,a CArG box and a Y box in U3 of JS13 LY19 genome was found to be deleted compared with that of exogenous ALV.Therefore,it is speculated that JS13 LY19 is generated by the recombination of JS11 C1 and endogenous ALV.Having endogenous ALV LTR and partial deletions of transcription regulatory elements may reduce the transcription capacity and pathogenicity of JS13 LY19.
引文
[1] 蔺文成,李昕键,严一铭,等.J亚群禽白血病病毒GD14A8株分离鉴定及遗传进化分析[J].中国兽医学报,2017,37(2):206-210.
[2] ZHAO P,DONG X,CUI Z.Isolation,identification,and gp85 characterization of a subgroup A avian leukosis virus from a contaminated live Newcastle disease virus vaccine,first report in China[J].Poult Sci,2014,93(9):2168-2174.
[3] PAYNE L N,NAIR V.The long view:40 years of avian leukosis research[J].Avian Pathol,2012,41(1):11-19.
[4] 王鑫,赵鹏,崔治中.我国地方品种鸡分离到的一个禽白血病病毒新亚群的鉴定[J].病毒学报,2012,28(6):609-613.
[5] LI X,LIN W,CHANG S,et al.Isolation,identification and evolution analysis of a novel subgroup of avian leukosis virus isolated from a local Chinese yellow broiler in South China[J].Arch Virol,2016,161(10):2717-2725.
[6] SHAO H,WANG L,SANG J,et al.Novel avian leukosis viruses from domestic chicken breeds in mainland China[J].Arch Virol,2017,162(7):2073-2076.
[7] 李久庆,刘强,郭雷,等.一株新亚群禽白血病病毒全基因组序列分析[J].畜牧兽医学报,2017,48(9):1718-1723.
[8] SU Q,LI Y,CUI Z,et al.The emerging novel avian leukosis virus with mutations in the pol gene shows competitive replication advantages both in vivo and in vitro[J].Emerging Microbes Infect,2018,7(1):117.
[9] ZHAO Z,RAO M,LIAO M,et al.Phylogenetic analysis and pathogenicity assessment of the emerging recombinant subgroup K of avian leukosis virus in south China[J].Viruses,2018,10(4):194.
[10] JOHNSON J A,HENEINE W.Characterization of endogenous avian leukosis viruses in chicken embryonic fibroblast substrates used in production of measles and mumps vaccines[J].J Virol,2001,75(8):3605-3612.
[11] BARBOSA T,ZAVALA G,CHENG S.Molecular characterization of three recombinant isolates of avian leukosis virus obtained from contaminated Marek’s disease vaccines[J].Avian Dis,2014,52(2):245-252.
[12] BROWN D W,ROBINSON H L.Influence of env and long terminal repeat sequences on the tissue tropism of avian leukosis viruses[J].J Virol,1988,62(12):4828-4831.
[13] RUDDELL A.Transcription regulatory elements of the avian retroviral long terminal repeat[J].Virology,1995,206(1):1-7.
[14] CHANG S W,HSU M F,WANG C H.Gene detection,virus isolation,and sequence analysis of avian leukosis viruses in Taiwan country chickens[J].Avian Dis,2013,57(2):172-177.
[15] OCHI A,OCHIAI K,KOBARA A,et al.Epidemiological study of fowl glioma-inducing virus in chickens in Asia and Germany[J].Avian Pathol,2012,41(3):299-309.
[16] MOTTA J V,CRITTENDEN L B,PURCHASE H G,et al.Low oncogenic potential of avian endogenous RNA tumor virus infection or expression[J].J National Cancer Institute,1975,55(3):685-689.
[17] CULLEN B R,RAYMOND K,JU G.Transcriptional activity of avian retroviral long terminal repeats directly correlates with enhancer activity[J].J Virol,1985,53(2):515-521.
[18] 赵子君,饶明章,陈建,等.K亚群禽白血病病毒5′ LTR序列及启动活性分析[J].畜牧兽医学报,2018,49(4):754-760.
[19] SEARS R O,SEALY L.Characterization of nuclear proteins that bind the EFII enhancer sequence in the Rous Sarcoma virus long terminal repeat[J].J Virol,1992,66(11):6338-6352.
[20] LANG A,FINCHAM V J,WYKEJ A.Factors influencing physiological variations in the activity of the Rous sarcoma virus long terminal repeat[J].Virology,1993,196:564-575.
[21] NAKAMURA S,OCHIAI K,OCHI A,et al.Cardiac pathology and molecular epidemiology by avian leukosis viruses in Japan[J].PLoS One,2014,9(1):e86546.
[2] ZHAO P,DONG X,CUI Z.Isolation,identification,and gp85 characterization of a subgroup A avian leukosis virus from a contaminated live Newcastle disease virus vaccine,first report in China[J].Poult Sci,2014,93(9):2168-2174.
[3] PAYNE L N,NAIR V.The long view:40 years of avian leukosis research[J].Avian Pathol,2012,41(1):11-19.
[4] 王鑫,赵鹏,崔治中.我国地方品种鸡分离到的一个禽白血病病毒新亚群的鉴定[J].病毒学报,2012,28(6):609-613.
[5] LI X,LIN W,CHANG S,et al.Isolation,identification and evolution analysis of a novel subgroup of avian leukosis virus isolated from a local Chinese yellow broiler in South China[J].Arch Virol,2016,161(10):2717-2725.
[6] SHAO H,WANG L,SANG J,et al.Novel avian leukosis viruses from domestic chicken breeds in mainland China[J].Arch Virol,2017,162(7):2073-2076.
[7] 李久庆,刘强,郭雷,等.一株新亚群禽白血病病毒全基因组序列分析[J].畜牧兽医学报,2017,48(9):1718-1723.
[8] SU Q,LI Y,CUI Z,et al.The emerging novel avian leukosis virus with mutations in the pol gene shows competitive replication advantages both in vivo and in vitro[J].Emerging Microbes Infect,2018,7(1):117.
[9] ZHAO Z,RAO M,LIAO M,et al.Phylogenetic analysis and pathogenicity assessment of the emerging recombinant subgroup K of avian leukosis virus in south China[J].Viruses,2018,10(4):194.
[10] JOHNSON J A,HENEINE W.Characterization of endogenous avian leukosis viruses in chicken embryonic fibroblast substrates used in production of measles and mumps vaccines[J].J Virol,2001,75(8):3605-3612.
[11] BARBOSA T,ZAVALA G,CHENG S.Molecular characterization of three recombinant isolates of avian leukosis virus obtained from contaminated Marek’s disease vaccines[J].Avian Dis,2014,52(2):245-252.
[12] BROWN D W,ROBINSON H L.Influence of env and long terminal repeat sequences on the tissue tropism of avian leukosis viruses[J].J Virol,1988,62(12):4828-4831.
[13] RUDDELL A.Transcription regulatory elements of the avian retroviral long terminal repeat[J].Virology,1995,206(1):1-7.
[14] CHANG S W,HSU M F,WANG C H.Gene detection,virus isolation,and sequence analysis of avian leukosis viruses in Taiwan country chickens[J].Avian Dis,2013,57(2):172-177.
[15] OCHI A,OCHIAI K,KOBARA A,et al.Epidemiological study of fowl glioma-inducing virus in chickens in Asia and Germany[J].Avian Pathol,2012,41(3):299-309.
[16] MOTTA J V,CRITTENDEN L B,PURCHASE H G,et al.Low oncogenic potential of avian endogenous RNA tumor virus infection or expression[J].J National Cancer Institute,1975,55(3):685-689.
[17] CULLEN B R,RAYMOND K,JU G.Transcriptional activity of avian retroviral long terminal repeats directly correlates with enhancer activity[J].J Virol,1985,53(2):515-521.
[18] 赵子君,饶明章,陈建,等.K亚群禽白血病病毒5′ LTR序列及启动活性分析[J].畜牧兽医学报,2018,49(4):754-760.
[19] SEARS R O,SEALY L.Characterization of nuclear proteins that bind the EFII enhancer sequence in the Rous Sarcoma virus long terminal repeat[J].J Virol,1992,66(11):6338-6352.
[20] LANG A,FINCHAM V J,WYKEJ A.Factors influencing physiological variations in the activity of the Rous sarcoma virus long terminal repeat[J].Virology,1993,196:564-575.
[21] NAKAMURA S,OCHIAI K,OCHI A,et al.Cardiac pathology and molecular epidemiology by avian leukosis viruses in Japan[J].PLoS One,2014,9(1):e86546.