羊口疮病毒疫苗株与野毒株VIL-10基因的比较分析

doi:10.16431/j.cnki.1671-7236.2018.09.003

《中国畜牧兽医》 ›› 2018, Vol. 45 ›› Issue (9): 2368-2376.doi: 10.16431/j.cnki.1671-7236.2018.09.003

羊口疮病毒疫苗株与野毒株VIL-10基因的比较分析

冯倩, 吴锦艳, 陈妍, 刘永杰, 张克山, 尚佑军, 刘湘涛

中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, 农业部畜禽病毒学重点开放实验室, 兰州 730046

收稿日期:2018-03-01 出版日期:2018-09-20 发布日期:2018-09-26
通讯作者: 尚佑军 E-mail:shangyoujun@caas.cn
作者简介:冯倩(1993-),女,山西临汾人,硕士生,研究方向:预防兽医学,E-mail:756058113@qq.com
基金资助:
国家重点研发计划课题（2017YFD0500903）；国家现代肉羊产业技术体系资助项目（CARS-39-04B）

Comparative Analysis of VIL-10 Gene Between the Vaccine and Wild Strains of Orf Virus

FENG Qian, WU Jinyan, CHEN Yan, LIU Yongjie, ZHANG Keshan, SHANG Youjun, LIU Xiangtao

Key Laboratory of Animal Virology of the Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Lanzhou 730046, China

Received:2018-03-01 Online:2018-09-20 Published:2018-09-26

摘要/Abstract

摘要：

试验旨在比较分析羊口疮病毒（orf virus，ORFV）VIL-10基因在疫苗株和野毒株之间的差异特征。参照GenBank中公布的ORFV NZ2株的VIL-10基因序列设计并合成1对特异性引物，分别以疫苗株和野毒株提取的基因组DNA为模板，采用PCR方法扩增ORFV的VIL-10基因全序列并进行测序，应用生物信息学相关软件分析基因的核苷酸、氨基酸变异情况及蛋白结构。结果显示，本试验测定的疫苗株和野毒株VIL-10基因核苷酸序列同源性为94.4%，差异主要是单个碱基的突变，其中疫苗株在132~134 bp核苷酸序列出现缺失；氨基酸序列同源性为92.5%，出现了15个氨基酸位点的突变，其中疫苗株第42位氨基酸天冬酰胺出现缺失；蛋白质在一级结构及理化性质、二级结构、三级结构、抗原表位参数及有无信号肽之间均存在一定程度的差异，而疫苗株和野毒株编码的蛋白质均无跨膜结构域。系统进化树分析结果表明，本试验测定的野毒株与疫苗株属于不同分支，遗传关系较远。研究结果提示，野毒株与疫苗株的VIL-10基因发生较明显的变异，这些变异可能与ORFV疫苗株的毒力致弱有关。

关键词: 羊口疮病毒(ORFV); 疫苗株; 野毒株; VIL-10基因; 比较分析

Abstract:

To analyze the different characteristics between VIL-10 gene from the vaccine and wild strains of orf virus (ORFV),a pair of specific primers was designed according to ORFV NZ2 strain VIL-10 gene sequence reported in GenBank.The genomic DNA was taken as templates extracted from the vaccine and wild strains of ORFV.The full lengths of VIL-10 gene were obtained by PCR and then sequenced.The variation of nucleic acids,amino acids and protein structures were predicted and analyzed using bioinformatics software.The results showed that VIL-10 gene from ORFV vaccine strain and wild strain shared 94.4% identity at nucleotide level and the main difference was a single base mutation,deletion in the nucleotide sequence which occurred at 132 to 134 bp.92.5% identity at amino acid level and a mutation of 15 amino acid sites occurred,of which the asparagine was missing at 42 amino acid position.There were some differences in primary structure and physical and chemical properties of protein,secondary structure,tertiary structure,physicochemical properties,epitope parameters and the presence or absence of signal peptide.However,no transmembrane domains were identified both in the vaccine and virulent strains of ORFV.Phylogenetic analysis revealed that the wild strain and the vaccine strain belonged to different branches and the genetic relationship was far away.The results suggested that the VIL-10 gene of wild virus strains and vaccine strains had obvious mutations,and these mutations might be related to the virulence of the vaccine strains.

Key words: orf virus (ORFV); vaccine strain; wild strain; VIL-10 gene; comparative analysis

中图分类号:

S852.65⁺4

冯倩, 吴锦艳, 陈妍, 刘永杰, 张克山, 尚佑军, 刘湘涛. 羊口疮病毒疫苗株与野毒株VIL-10基因的比较分析[J]. 《中国畜牧兽医》, 2018, 45(9): 2368-2376.

FENG Qian, WU Jinyan, CHEN Yan, LIU Yongjie, ZHANG Keshan, SHANG Youjun, LIU Xiangtao. Comparative Analysis of VIL-10 Gene Between the Vaccine and Wild Strains of Orf Virus[J]. , 2018, 45(9): 2368-2376.

参考文献

[1] 殷震,刘景华.动物病毒学[M].第2版.北京:科学出版社,1997.YIN Z,LIU J H.Animal Virology[M].2nd Edition.Beijing:Science Press,1997.(in Chinese)
[2] TEDLA M,BERHAN N,MOLLA W,et al.Molecular identification and investigations of contagious ecthyma (orf virus) in small ruminants,North West Ethiopi[J].BMC Veterinary Research,2018,14(1):13.
[3] NANDI S,DE U K,CHOWDHURY S.Current status of contagiousecthyma or orf disease in goat and sheep-A global perspective[J].Small Ruminant Research,2011,96(2):73-82.
[4] HOSAMANI M,SCAGLIARINI A,BHANUPRAKASH V,et al.Orf:An update on current research and future perspectives[J].Expert Review of Anti-infective Therapy,2009,7(7):879-893.
[5] BERGQVIST C,KURBAN M,ABBAS O.Orf virus infection[J].Reviews in Medical Virology, 2017,27(4):e1932.
[6] TRYLAND M,BECKMEN K B,BUREK-HUNTINGTON K A,et al.Orf virus infection in Alaskan mountain goats,Dall's sheep,muskoxen,caribou and Sitka Black-tailed deer[J].Acta Veterinaria Scandinavica,2018,60(1):12.
[7] LI W,NING Z,HAO W,et al.Isolation and phylogenetic analysis of orf virus from the sheep herd outbreak in Northeast China[J].BMC Veterinary Research,2012,8(1):229.
[8] HAIG D M.Orf virus infection and host immunity[J].Current Opinion in Infectious Diseases,2006,19(2):127-131.
[9] CHEN H,LI W,KUANG Z,et al.The whole genomic analysis of orf virus strain HN3/12 isolated from Henan province,central China[J].BMC Veterinary Research,2017,13(1):260.
[10] YU Y,TAN Q,ZHAO W,et al.Characterization of anorf virus isolate from an outbreak in Heilongjiang province,China[J].Archives of Virology,2017,162(10):3143-3149.
[11] 王光祥,尚佑军,吕占禄,等.羊口疮病毒B2L蛋白二级结构及其细胞表位的预测[J].中国兽医科学,2012,42(11):1133-1138.WANG G X,SHANG Y J,LV Z L,et al.Prediction of secondary structure and cell epitopes of B2L protein of orf virus[J].China Veterinary Science,2012,42(11):1133-1138.(in Chinese)
[12] ROBINSON A J,ELLIS G,BALASSU T.The genome of orf virus:Restriction endonuclease analysis of viral DNA isolated from lesions of orf in sheep[J].Archives of Virology,1982,71(1):43-55.
[13] TSAI S M,CHAN K W,HSU W L,et al.Development of a loop-mediated isothermal amplification for rapid detection of orf virus[J].Journal of Virological Methods,2009,157(2):200-204.
[14] 李慧霞,朱学亮,骆学农.羊口疮病的研究进展[J].中国预防兽医学报,2013,35(5):426-430.LI H X,ZHU X L,LUO X N.Research progress on orf virus[J].Chinese Journal of Preventive Veterinary Medicine,2013,35(5):426-430.
[15] CHAN A,BAIRD M,MERCERA A,et al.Maturation and function of human dendritic cells are inhibited by orf virus-encoded interleukin-10[J].Journal of General Virology,2006,87(11):3177-3181.
[16] IMLACH W,MCCAUGHAN C A,Mercer A A,et al.Orf virus-encoded interleukin-10 stimulates the proliferation of murine mast cells and inhibits cytokine synthesis in murine peritoneal macrophages[J].Journal of General Virology,2002,83(5):1049-1058.
[17] DING Y,QIN L,KOTENKO S V,et al.A single amino acid determines the immunostimulatory activity of interleukin 10[J].Journal of Experimental Medicine,2000,191(2):213-224.
[18] DEMIRASLAN H,DINC G,DOGANAY M.Anoverview of orf virus infection in humans and animals[J].Recent Patents on Anti-infective Drug Discovery,2017,12(1):21-30.
[19] 刘方,赵玄多,安贝,等.羊口疮病毒凤翔株ORFV121和vIL-10基因的生物信息学分析[J].动物医学进展,2014,35(7):52-58.LIU F,ZHAO X D,AN B,et al.Bioinformatics analyses ORFV121 gene and vIL-10 gene of ORFV-FX0910 strain[J].Progress in Veterinary Medicine,2014,35(7):52-58.(in Chinese)
[20] 于永忠,吴志军,朱战波,等.羊口疮病毒分子特征与免疫逃逸策略[J].病毒学报,2012,28(3):278-284.YU Y Z,WU Z J,ZHU Z B,et al.Molecular characteristics and immune evasion strategies of ORFV:A review[J].Journal of Virology,2012,28(3):278-284.(in Chinese)
[21] BENNETT J R,LATEEF Z,FLEMING S B,et al.Orf virus IL-10 reduces monocyte,dendritic cell and mast cell recruitment to inflamed skin[J].Virus Research,2016,213:230-237.
[22] ROJAS J M,AVIA M,MARTIN V,et al.IL-10:A multifunctional cytokine in viral infections[J].Journal of Immunology Research,2017,2017(4):6104054.
[23] KRUSE N,WEBER O.Selective induction of apoptosis in antigen-presenting cells in mice by Parapoxvirus ovis[J].Journal of Virology,2001,75(10):4699-4704.
[24] 张海,周碧君,杨忠成,等.牛乳头状瘤病毒贵州株L1基因克隆及生物信息学分析[J].中国畜牧兽医,2016,43(11):2834-2843.ZHANG H,ZHOU B J,YANG Z C,et al.Cloning and bioinformatic analysis of L1 gene of BPV from Guizhou province[J].China Animal Husbandry & Veterinary Medicine,2016,43(11):2834-2843.(in Chinese)

羊口疮病毒疫苗株与野毒株VIL-10基因的比较分析

Comparative Analysis of VIL-10 Gene Between the Vaccine and Wild Strains of Orf Virus

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