鸭源H6N6亚型禽流感病毒贵州株NA基因序列分析
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摘要
为了解H6N6亚型禽流感病毒贵州株N6基因生物学特性,根据GenBank上禽流感病毒N6基因序列设计特异性引物,对贵州地区分离的H6N6亚型禽流感病毒贵州株进行N6基因RT-PCR扩增与克隆测序分析,结果显示,4株H6N6亚型禽流感病毒贵州株N6基因编码区均为1 380bp,可编码459个氨基酸,其中从第175-207位缺失33个核苷酸,即编码蛋白颈部存在11个氨基酸缺失(TNSTTTIINNN);4株H6N6亚型禽流感病毒贵州株N6基因核苷酸同源性为96.0%～99.1%,其中A/duck/Guizhou/01/2017(H6N6)与A/duck/Guizhou/02/2017(H6N6)为同一支,A/duck/Guizhou/03/2017(H6N6)和A/duck/Guizhou/04/2017(H6N6)为另一同支;N6亚型毒株与其他非N6亚型毒株明显位于2个分支上。这些结果为贵州省禽流感分子流行病学研究提供了基础资料。
To investgate the biological charateristics of N6 gene of avian influenza virus of H6N6 subtype in Guizhou province,the specific primers were designed on the basis of the N6 gene of avian influenza virus in GenBank,and reverse transcription-polymerase chain reaction(RT-PCR)was used to amplify the fulllength cDNAs of ducks avian influenza virus(AIV).The results showed that the full-length of NA genes were 1 464 bp and its open reading frame(ORF)was 1 380 bp,which encoded 459 amino acids.The NA gene also had 33 nucleotides deletion from position 175 to 207 at the N6 gene stalk.Therefore,the ORF encoded 459 amino acids which deleted 11 amino acids(TNSTTTIINN).The homology of N6 genes was96.0%-99.1%,A/duck/Guizhou/01/2017(H6N6)and A/duck/Guizhou/02/2017(H6N6)located in the same branch,while A/duck/Guizhou/03/2017(H6N6)and A/duck/Guizhou/04/2017(H6N6)located in another branch.Fathermore,N6 subtype strains and other non-N6 subtype strains are prosioned on the two branches obviously.These results could provide some basic data for studying the molecular epidemiology of avian influenza in Guizhou province.
To investgate the biological charateristics of N6 gene of avian influenza virus of H6N6 subtype in Guizhou province,the specific primers were designed on the basis of the N6 gene of avian influenza virus in GenBank,and reverse transcription-polymerase chain reaction(RT-PCR)was used to amplify the fulllength cDNAs of ducks avian influenza virus(AIV).The results showed that the full-length of NA genes were 1 464 bp and its open reading frame(ORF)was 1 380 bp,which encoded 459 amino acids.The NA gene also had 33 nucleotides deletion from position 175 to 207 at the N6 gene stalk.Therefore,the ORF encoded 459 amino acids which deleted 11 amino acids(TNSTTTIINN).The homology of N6 genes was96.0%-99.1%,A/duck/Guizhou/01/2017(H6N6)and A/duck/Guizhou/02/2017(H6N6)located in the same branch,while A/duck/Guizhou/03/2017(H6N6)and A/duck/Guizhou/04/2017(H6N6)located in another branch.Fathermore,N6 subtype strains and other non-N6 subtype strains are prosioned on the two branches obviously.These results could provide some basic data for studying the molecular epidemiology of avian influenza in Guizhou province.
引文
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[12]China CDC(2014).Avian influenza A(H5N6)virus was detected in a respiratory tract sample from apatient who died of severe pneumonia in Sichuan[EB/OL].[2015-12-21].http://www.Chinacdc.cn/mtdx/cr bxx/201405/t20140507-96416.htm.
[13]Bi Yuhai,Chen Quanjiao,Wang Qianli,et al.Genesis,evolution and prevalence of H5N6avian influenza viruses in China[J].Cell Host&Microbe,2016,20(6):810!821.
[14]任丽伟,丁爱军,杨海燕,等.禽流感病毒NA基因稀有密码子分析及其原核表达[J].中国家禽,2009,31(21):18-21.
[15]万春和,傅光华,施少华,等.一株番鸭源N6亚型禽流感病毒神经氨酸酶基因的克隆及序列分析[J].中国动物传染病学报,2010,18(2):28-33.
[16]王新卫,詹爱军.禽流感病毒的宿主特异性和致病性分子基础[J].长江大学学报:自科版,2006(2):161-164.
[17]Zhao G,Lu X,Gu X,et al.Molecular evolution of the H6subtype influenza A viruses from poultry in eastern China from2002to 2010[J].J Virol,2011,8(1):1-12.
[18]Jiao P,Yuan R,Wei L,et al.Complete genomic sequence of a novel natural recombinant H6N2influenza virus from chickens in Guangdong,southern China[J].J Virol,2012,86(14):7717-7718.
[19]谭力凯,张民泽,邵振文,等.1株禽源猪流感H6N6病毒的反向遗传系统的建立[J].中国兽医杂志,2014,50(10):6-8.
[20]Zhang G,Kong W,Qi W,et al.Identification of an H6N6swine influenza virus in southern China[J].Infect Genet Evol,2011,11(5):1174-1177.
[2]Pushko P,Tretyakova I,Hidajat R,et al.Virus-like particles displaying H5,H7,H9hemagglutinins and N1neuraminidase elicit protective immunity to heterologous avian influenza viruses in chickens[J].Virology,2017(501):176-182.
[3]Li Q,Sun X,Li Z,et al.Structural and functional characterization of neuraminidase-like molecule N10derived from bat influenza Avirus[J].Proc Natl Acal Sci USA,2012,109(46):18897-18902.
[4]袁润余,康银峰,王峥,等.高通量测序H6N6亚型禽流感病毒及遗传进化分析[J].华南预防医学,2016,42(4):301-308.
[5]仇保丰,胡顺林,刘武杰,等.六株H4N6亚型水禽禽流感病毒分离株NA基因的进化分析[J].中国家禽,2009,31(15):13-15.
[6]李昀海,赵焕云,张文东,等.云南2株H4N6亚型禽流感病毒株的分离与鉴定[J].畜牧与兽医,2011,43(6):12-17.
[7]解林红,高晓龙,耿海东,等.H4N6亚型禽流感病毒分离与氨基酸序列测定[J].中国病原生物学杂志,2015,10(5):385-388.
[8]莫胜兰,郑敏,陈义祥,等.广西鸭源H6N6亚型禽流感病毒HA和NA基因的序列分析[J].动物医学进展,2011,32(12):11-15.
[9]宋跃君,谭丹,邓国华,等.湖南省洞庭湖区5株H6N6亚型禽流感毒株HA、NA基因的遗传进化分析[J].中国动物检疫,2016,33(1):9-12.
[10]陈佳琪.鸭源H6N6亚型禽流感病毒的分离鉴定与致病性分析[D].贵州贵阳:贵州大学,2016.
[11]蒋文明,陈继明.我国高致病性禽流感的流行与防控[J].中国动物检疫,2015,32(6):5-9.
[12]China CDC(2014).Avian influenza A(H5N6)virus was detected in a respiratory tract sample from apatient who died of severe pneumonia in Sichuan[EB/OL].[2015-12-21].http://www.Chinacdc.cn/mtdx/cr bxx/201405/t20140507-96416.htm.
[13]Bi Yuhai,Chen Quanjiao,Wang Qianli,et al.Genesis,evolution and prevalence of H5N6avian influenza viruses in China[J].Cell Host&Microbe,2016,20(6):810!821.
[14]任丽伟,丁爱军,杨海燕,等.禽流感病毒NA基因稀有密码子分析及其原核表达[J].中国家禽,2009,31(21):18-21.
[15]万春和,傅光华,施少华,等.一株番鸭源N6亚型禽流感病毒神经氨酸酶基因的克隆及序列分析[J].中国动物传染病学报,2010,18(2):28-33.
[16]王新卫,詹爱军.禽流感病毒的宿主特异性和致病性分子基础[J].长江大学学报:自科版,2006(2):161-164.
[17]Zhao G,Lu X,Gu X,et al.Molecular evolution of the H6subtype influenza A viruses from poultry in eastern China from2002to 2010[J].J Virol,2011,8(1):1-12.
[18]Jiao P,Yuan R,Wei L,et al.Complete genomic sequence of a novel natural recombinant H6N2influenza virus from chickens in Guangdong,southern China[J].J Virol,2012,86(14):7717-7718.
[19]谭力凯,张民泽,邵振文,等.1株禽源猪流感H6N6病毒的反向遗传系统的建立[J].中国兽医杂志,2014,50(10):6-8.
[20]Zhang G,Kong W,Qi W,et al.Identification of an H6N6swine influenza virus in southern China[J].Infect Genet Evol,2011,11(5):1174-1177.