2017年不同地区黄鸡中鸡传染性支气管炎病毒分离株N基因遗传进化分析
|
摘要
为分析2017年国内不同地区黄羽肉鸡中传染性支气管炎病毒(IBV)核衣壳蛋白基因(N基因)的遗传进化规律,本研究在2017年从广西、云南、山东等地共分离到16株IBV株,并对各分离株进行N基因扩增、测序及分析。结果显示:16株分离株N基因全长均为1 230 bp,不存在碱基缺失和插入。同源性分析显示:16株IBV分离株之间核苷酸和推导氨基酸同源性分别为85.9%～99.9%和89%～99.8%。进化分析显示:16株分离株中12株属于QX-Like型,3株属于Mass-Like型,1株属于4/91-Like型。以上结果表明:IBV N基因在我国部分地区主要以QX-Like型为主,已形成独立进化群,同时存在基因突变及重组的现象。本研究首次对不同地区黄羽肉鸡IBV N基因遗传进化进行了分析,阐明了该基因分子流行病学特征。
This study was aimed to investigate the variations in the nucleocapsid(N)gene of infectious bronchitis viruses(IBVs)isolated in some areas of China.Sixteen IBVs were isolated from Guangxi,Yunnan and Shandong provinces in 2017.The N genes of the various IBV isolates were amplified by RT-PCR,cloned and sequenced.The results showed that the ORFs of all the sixteen IBVs were 1,230bp,encoding 409 amino acids.The IBV isolates shared nucleotide sequence similarity between 85.9%and 99.9%,the deduced amino acid sequence similarity between 89%and 99.8%.Phylogenetic analysis of the deduced N nucleotide sequences of the 16 IBVs clustered into three published IBV clusters,among which 12 isolates belonged to the branch of QX-Like cluster,3 isolates belonged to the branch of Mass-Like cluster,1 isolate belonged to the branch of 4/91-Like cluster.These results suggested that the QX-Like cluster isolates were the dominate gene type in some areas of China and formed their own independent evolution.At the same time the mutations had occurred in the nucleocapsid protein gene of Chinese IBVs isolated in 2017.This study for the first time to investigate the variations in the N gene of IBV isolated in yellow chickens,and illustrates the N gene molecular epidemiological characteristics.
This study was aimed to investigate the variations in the nucleocapsid(N)gene of infectious bronchitis viruses(IBVs)isolated in some areas of China.Sixteen IBVs were isolated from Guangxi,Yunnan and Shandong provinces in 2017.The N genes of the various IBV isolates were amplified by RT-PCR,cloned and sequenced.The results showed that the ORFs of all the sixteen IBVs were 1,230bp,encoding 409 amino acids.The IBV isolates shared nucleotide sequence similarity between 85.9%and 99.9%,the deduced amino acid sequence similarity between 89%and 99.8%.Phylogenetic analysis of the deduced N nucleotide sequences of the 16 IBVs clustered into three published IBV clusters,among which 12 isolates belonged to the branch of QX-Like cluster,3 isolates belonged to the branch of Mass-Like cluster,1 isolate belonged to the branch of 4/91-Like cluster.These results suggested that the QX-Like cluster isolates were the dominate gene type in some areas of China and formed their own independent evolution.At the same time the mutations had occurred in the nucleocapsid protein gene of Chinese IBVs isolated in 2017.This study for the first time to investigate the variations in the N gene of IBV isolated in yellow chickens,and illustrates the N gene molecular epidemiological characteristics.
引文
[1]Boursnell M E,Binns M M,Foulds I J,et al.Sequences of the nucleocapsid genes from two strains of avian infectious bronchitis virus[J].J Gen Virol,1985,66(Pt3):573-580.
[2]Sapats S I,Ashton F,Wright P J,et al.Novel variation in the N protein of avian infectious bronchitis virus[J].Virology,1996,226(2):412-417.
[3]Chen Hong-ying,Gill A,Dove B K,et al.Mass spectroscopic characterization of the coronavirus infectious bronchitis virus nucleoprotein and elucidation of the role of phosphorylation in RNA binding by using surface plasmon resonance[J].J Virol,2005,79(2):1164-1179.
[4]Zhou Ming-long,Williams A K,Chung S,et al.The infectious bronchitis virus nucleocapsid protein binds RNA sequences in the 3'teminus of the genome[J].Virology,1996,217(1):191-199.
[5]Yu Dan,Han Zong-xi,Xu Jia,et al.A novel B-cell epitope of avian infectious bronchitis virus N protein[J].Viral Immunol,2010,23(2):189-199.
[6]Boots A M,Benaissa-Trouw B J,Hesselink W,et al.Induction of anti-viral immune responses by immunization with recombinant-DNA encoded avian coronavirus nucleocapsid protein[J].Vaccine,1992,10(2):119-124.
[7]于冬玲,谭春萍,张艳雯.广西2013年~2015年鸡传染性支气管炎病毒S1基因分子流行病学分析[J].动物医学进展,2016,37(8):19-23.
[8]Xu Gang,Liu Xiao-yu,Zhao Ye,et al.Characterization and analysis of an infectious bronchitis virus strain isolated from southern China in 2013[J].Virol J,2016,13(1):1-9.
[9]盛晓丹,徐怀英,刘霞,等.传染性支气管炎病毒S1和N基因遗传变异频率的分析[J].畜牧兽医学报,2015,46(6):989-997.
[10]Mo Mei-lan,Li Meng,Huang Bai-cheng,et al.Molecular characterization of major structural protein genes of avian coronavirus infectious bronchitis virus isolates in southern China[J].Viruses,2013,5(12):3007-3020.
[11]孙晓云.鸡传染性支气管炎病毒H120型M、N基因的免疫原性研究[D].上海:上海海洋大学,2016.
[2]Sapats S I,Ashton F,Wright P J,et al.Novel variation in the N protein of avian infectious bronchitis virus[J].Virology,1996,226(2):412-417.
[3]Chen Hong-ying,Gill A,Dove B K,et al.Mass spectroscopic characterization of the coronavirus infectious bronchitis virus nucleoprotein and elucidation of the role of phosphorylation in RNA binding by using surface plasmon resonance[J].J Virol,2005,79(2):1164-1179.
[4]Zhou Ming-long,Williams A K,Chung S,et al.The infectious bronchitis virus nucleocapsid protein binds RNA sequences in the 3'teminus of the genome[J].Virology,1996,217(1):191-199.
[5]Yu Dan,Han Zong-xi,Xu Jia,et al.A novel B-cell epitope of avian infectious bronchitis virus N protein[J].Viral Immunol,2010,23(2):189-199.
[6]Boots A M,Benaissa-Trouw B J,Hesselink W,et al.Induction of anti-viral immune responses by immunization with recombinant-DNA encoded avian coronavirus nucleocapsid protein[J].Vaccine,1992,10(2):119-124.
[7]于冬玲,谭春萍,张艳雯.广西2013年~2015年鸡传染性支气管炎病毒S1基因分子流行病学分析[J].动物医学进展,2016,37(8):19-23.
[8]Xu Gang,Liu Xiao-yu,Zhao Ye,et al.Characterization and analysis of an infectious bronchitis virus strain isolated from southern China in 2013[J].Virol J,2016,13(1):1-9.
[9]盛晓丹,徐怀英,刘霞,等.传染性支气管炎病毒S1和N基因遗传变异频率的分析[J].畜牧兽医学报,2015,46(6):989-997.
[10]Mo Mei-lan,Li Meng,Huang Bai-cheng,et al.Molecular characterization of major structural protein genes of avian coronavirus infectious bronchitis virus isolates in southern China[J].Viruses,2013,5(12):3007-3020.
[11]孙晓云.鸡传染性支气管炎病毒H120型M、N基因的免疫原性研究[D].上海:上海海洋大学,2016.