猪流行性腹泻病毒山东分离株SD-2016的S基因序列分析
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摘要
为研究本实验室分离的SD-2016分离株S基因序列特点,设计扩增猪流行性腹泻病毒(PEDV)S基因的2对引物,将扩增出的目的片段与克隆载体连接后测序,利用分子生物学软件对目的基因的核苷酸序列和氨基酸序列进行同源性比对分析。结果:SD-2016毒株与CV777、LZC等GI群的毒株在核苷酸序列同源性比对结果为93.8%~95.0%,氨基酸序列同源性比对结果在92.5%~94.1%。绘制系统发育进化树发现,SD-2016株与中国广东分离株GD12、湖南分离株HUN在一个分支上,亲缘关系较近。对S基因的中和表位进行分析发现主要的氨基酸位点突变集中于抗原表位区COE区域(第499-638位氨基酸)和SS6(第764-771位氨基酸),分别有6个位点和2个位点的突变。通过对SD-2016分离株S基因的分析,为以后猪流行性腹泻病的基因工程疫苗研制提供参考。
In order to study the characteristics of S gene sequence of SD-2016 which was isolated in our laboratory,two pairs of primers for amplification of PEDV S gene were designed. The amplified fragments were cloned into the cloning vector and sequenced. Homology analysis of the nucleotides and amino acid sequences of the target gene showed that the nucleotide sequence homology between the SD-2016 strain and the GI group of CV777 and LZC strains was 93. 8% ~ 95. 0%,and the amino acid sequence homology was 92. 5% ~ 94. 1%. Phylogenetic analysis showed that the SD-2016 strain was clustered with Guangdong GD12 and Hunan HUN in one branch. Analysis of the neutralizing epitopes of the S gene revealed that the major amino acid site mutations existed in the COE region and the SS6 region,with six and two site mutations,respectively. Analysis of the S gene of the SD-2016 isolate provided a theoretical basis for future development of genetically engineered vaccine for swine epidemic diarrhea.
In order to study the characteristics of S gene sequence of SD-2016 which was isolated in our laboratory,two pairs of primers for amplification of PEDV S gene were designed. The amplified fragments were cloned into the cloning vector and sequenced. Homology analysis of the nucleotides and amino acid sequences of the target gene showed that the nucleotide sequence homology between the SD-2016 strain and the GI group of CV777 and LZC strains was 93. 8% ~ 95. 0%,and the amino acid sequence homology was 92. 5% ~ 94. 1%. Phylogenetic analysis showed that the SD-2016 strain was clustered with Guangdong GD12 and Hunan HUN in one branch. Analysis of the neutralizing epitopes of the S gene revealed that the major amino acid site mutations existed in the COE region and the SS6 region,with six and two site mutations,respectively. Analysis of the S gene of the SD-2016 isolate provided a theoretical basis for future development of genetically engineered vaccine for swine epidemic diarrhea.
引文
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[10]卢冰霞,秦毅斌,何颖,等.2011—2014年广西猪流行性腹泻病毒检测及其M基因的序列分析[J].中国畜牧兽医,2015,42(3):549-557.
[11]韩莎莎,张玉娟,时庆贺,等.河南省2014-2015年猪流行性腹泻病毒流行株遗传进化分析[J].中国兽医学报,2017,12(6):976-983.
[2]杜以军,吴家强,刘纪玉,等.当前山东省猪群疫病的流行特点及防控对策[J].中国猪业,2014,9(1):37-39.
[3]Wang X M,Niu B B,Yan H,et al.Complete genome sequence of a variant porcine epidemic diarrhea virus strain isolated in central China[J].Genome Announc,2013,1(1).pii:e00243-12.
[4]Chen J,Wang,Shi H,et al.Molecular epidemiology of porcine epidemic siarrhea virus in China[J].Arch Virol,2010,155(9):1471-1476.
[5]Suo Siqingaowa,Ren Yudong,Li Guangxing,et al.Immune responses induced by DNA vaccines bearing spike gene of PEDV combined with porcine IL-18[J].Virus Res,2012(2)167(2):259-266.
[6]Millet J K,Whittaker G R.Host cell proteases:critical determinants of coronavirus tropism and pathogenesis[J].Virus Res,2015,202:120-134.
[7]王隆柏,林裕胜,车勇良,等.猪流行性腹泻病毒S、N和0RF3基因的遗传变异分析[J].畜牧兽医学报,2014,45(1):1830-1836.
[8]苏运芳,孙彦刚,刘运超,等.猪流行性腹泻疫苗研究进展[J].畜牧与兽医,2016,48(2):134-138.
[9]吴学敏,陈如敬,王隆柏,等.猪流行性腹泻病毒RT-PCR检测方法建立及初步应用研究[J].中国农学通报,2012,28(26):59-62.
[10]卢冰霞,秦毅斌,何颖,等.2011—2014年广西猪流行性腹泻病毒检测及其M基因的序列分析[J].中国畜牧兽医,2015,42(3):549-557.
[11]韩莎莎,张玉娟,时庆贺,等.河南省2014-2015年猪流行性腹泻病毒流行株遗传进化分析[J].中国兽医学报,2017,12(6):976-983.