Prokaryotic expression and bioinformatics analysis of orf virus Anhui strain 023 gene

doi:10.3969/j.issn.1004-1524.2019.01.04

›› 2019, Vol. 31 ›› Issue (1): 30-38.DOI: 10.3969/j.issn.1004-1524.2019.01.04

• OAnimal Science • Previous Articles Next Articles

Prokaryotic expression and bioinformatics analysis of orf virus Anhui strain 023 gene

ZHANG Gaofeng¹, YANG Kankan¹, ZHANG Qian¹, WANG Yuanhong¹, YU Zhaorong¹, ZHANG Xueqi¹, LU Zhimin¹, LIU Zimin¹, LI Yongdong^2,*, WANG Yong^1,*

1. College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China;
2. Ningbo Municipal Center for Disease Control and Prevention, Ningbo 315010, China

Received:2018-05-28 Online:2019-01-25 Published:2019-04-09

Abstract

Abstract: Our study was aimed at cloning and expressing the 023 gene of orf virus (ORFV) Anhui strain, and predicting its biological characteristics. The 023 gene was amplified by PCR, then successfully expressed using a prokaryotic vector. Then, the expressed protein was purified, and identified by SDS-PAGE and Western-blot. In addition, the physicochemical properties, secondary structure, signal peptide, transmembrane domain of the protein was analyzed by bioinformatics software. The results showed that the 023 gene of ORFV Anhui strain was 819 bp in length which could encode 272 amino acids. The size of expressed protein was approximately 42 ku, which was mainly expressed in inclusion body form with good reactogenicity. The results from bioinformatics analysis showed that the protein was a hydrophilic stable protein. There were 27.57% α-helix (h), 17.28% extended chain (e), 12.13% β-turn (t), 43.01% random coli (c). There was no signal peptide region and transmembrane domain. Maybe, it had 30 phosphorylation sites. Moreover, there were four B cell epitopes, three CTL epitopes and three Th epitopes. In conclusion, our study could provide a reference for further study of the structure and function of ORFV 023 gene.

Key words: orf virus, 023 gene, cloning and expression, bioinformatics

CLC Number:

S855.3

ZHANG Gaofeng, YANG Kankan, ZHANG Qian, WANG Yuanhong, YU Zhaorong, ZHANG Xueqi, LU Zhimin, LIU Zimin, LI Yongdong, WANG Yong. Prokaryotic expression and bioinformatics analysis of orf virus Anhui strain 023 gene[J]. , 2019, 31(1): 30-38.

References

[1] SPYROU V, VALIAKOS G.Orf virus infection in sheep or goats[J]. Veterinary Microbiology, 2015, 181(1/2): 178-182.
[2] KUMAR N, WADHWA A, CHAUBEY K K, et al.Isolation and phylogenetic analysis of an orf virus from sheep in Makhdoom, India[J]. Virus Genes, 2014, 48(2): 312-319.
[3] ZHANG K, LIU Y, KONG H, et al.Comparison and phylogenetic analysis based on the B2L gene of orf virus from goats and sheep in China during 2009-2011[J]. Archives of Virology, 2014, 159(6):1475-1479.
[4] Al-QATTAN M M. Orf infection of the hand[J]. Journal of Hand Surgery, 2011, 36(11): 1855-1858.
[5] FLEMING S B, WISE L M, MERCER A A.Molecular genetic analysis of orf virus: a poxvirus that has adapted to skin[J]. Viruses, 2015, 7(3):1505-1539.
[6] 王勇, 赵玉洁, 杨侃侃, 等. 口疮病毒020基因的克隆、表达及多抗制备[J]. 安徽农业大学学报, 2017, 44(5): 780-783.
WANG Y, ZHAO Y J, YANG K K, et al.Cloning and expression of the Orf virus 020 gene and its polyclonal antibody preparation[J].Journal of Anhui Agricultural University, 2017, 44(5): 780-783.(in Chinese with English abstract)
[7] 闫丰超, 邵佳, 窦永喜. 羊口疮病毒分子生物学的研究进展[J]. 中国兽医科学, 2013(1):103-109.
YAN F C, SHAO J, DOU Y X.Progress in molecular biology of orf virus[J]. Chinese Veterinary Science, 2013(1):103-109.(in Chinese with English abstract)
[8] 葛士坤, 张凯照, 鲁荣, 等. 2006—2016年中国羊口疮病毒的遗传演化分析[J]. 中国畜牧兽医, 2018,45(2):439-447.
GE S K, ZHANG K Z, LU R, et al.Genetic and phylogenetic analysis of orf virus isolated in China from 2006 to 2016[J]. China Animal Husbandry & Veterinary Medicine, 2018,45(2):439-447. (in Chinese with English abstract)
[9] SPYROU V, VALIAKOS G.Orf virus, infection in sheep or goats[J]. Veterinary Microbiology, 2015, 181(1/2):178-182.
[10] 成伟伟, 张克山, 刘永杰,等. 羊传染性脓疱病毒基因组结构和主要基因功能研究进展[J]. 动物医学进展, 2014, 35(7):82-85.
CHENG W W, ZHANG K S, LIU Y J, et al.Advance in genome structure and major gene function of orf virus[J].Progress in Veterinary Medicine, 2014,35(7):82-85. (in Chinese with English abstract)
[11] FRIEBE A, FRIEDERICHS S, SCHOLZ K, et al.Characterization of immunostimulatory components of orf virus (parapoxvirus ovis)[J]. Journal of General Virology, 2011, 92(Pt 7):1571-1584.
[12] WANG Y, YANG K, BAI C, et al.Development of a SYBR Green Ⅰ real-time PCR for the detection of the orf virus[J]. AMB Express, 2017, 7(1):21.
[13] YU Y, TAN Q, ZHAO W, et al.Characterization of an orf virus isolate from an outbreak in Heilongjiang province, China[J]. Archives of Virology, 2017,162(10):1-7.
[14] 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.
[15] CHI X, ZENG X, LUO S.Diagnosis and phylogenetic analysis of a multifocal cutaneous orf virus with mixed bacterial infection outbreak in goats in Fujian province,China[J]. Archives of Virology, 2017(10):1-10.
[16] 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, LYU Z L, et al.Prediction of secondary structure and cell epitopes of B2L protein of orf virus[J].Chinese Veterinary Science, 2012,42(11):1133-1138. (in Chinese with English abstract)
[17] 黄宇良, 汪立平, 陆克文, 等. 片球菌素pedA基因的原核表达、纯化及生物信息学分析[J].生物技术通报,2017,33(12):144-150.
HUANG Y L, WANG L P, LU K W, et al.Prokaryotic expression, purification and bioinformatics analysis of pediocin pedA gene[J].Biotechnology Bulletin, 2017,33(12):144-150. (in Chinese with English abstract)
[18] BHASIN M, RAGHAVA G P.Prediction of CTL epitopes using QM, SVM and ANN techniques[J]. Vaccine, 2004, 22(23/24):3195-3204.

Prokaryotic expression and bioinformatics analysis of orf virus Anhui strain 023 gene

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	ZHAO Xiuping, WANG Shuang, YAN Xingyi, DUAN Qiang, ZHANG Shuai, CHEN Yongsheng, LI Guorui. Expression, purification and bioinformatics analysis of Magnaporthe oryzae MGG-01005 [J]. Acta Agriculturae Zhejiangensis, 2021, 33(3): 470-478.
[2]	LIANG Liqin, YANG Rui, GAO Gang. Bioinformatics analysis of StUOXs gene family in potato [J]. , 2020, 32(9): 1523-1532.
[3]	WU Jia, CHEN Lang, JIANG Tao, HUANG Guoming, LI Zhuo, LI Yaodong, ZHANG Li, LIU Lixia. Genetic polymorphism screening of CSF3 gene in dairy cow and its bioinformatics analysis [J]. , 2020, 32(6): 986-993.
[4]	WANG Yuanhong, XING Xue, LI Chuanfeng, ZHU Jie, WANG Yong, LIU Guangqing. Bioinformatics analysis and prokaryotic expression of FIPV AH1905 strain N gene [J]. , 2020, 32(3): 406-414.
[5]	WANG Xiaopeng, ZHAO Liang, LIU Zimin, BAI Caixia, YANG Kankan, ZHANG Da, SUN Pei, JIANG Shudong, LI Yongdong, WANG Yong. Prokaryotic expression and bioinformatics analysis of PPV7 Cap gene [J]. , 2020, 32(2): 200-209.
[6]	TAN Guanghui, QIN Yuanyu, LI Jiezhang, WU Lei, YUE Yong, ZHANG Yiyu. Single nucleotide polymorphism screening and bioinformatics analysis of ATF4 gene in Cherry Valley ducks [J]. , 2020, 32(2): 218-225.
[7]	QIN Ling, ZHANG Xin, RONG Chunxiao, MO Chuanyuan, FAN Lu, YAN Jie, MENG Ying, ZHANG Manrang. Identification and expression analysis of polyamine oxidase (PAO) gene family in apple [J]. , 2020, 32(2): 262-273.
[8]	LIU Jialin, LIU Shili, JIANG Wenping, CHENG Shun, CHI Meili, ZHENG Jianbo, JIA Yongyi, ZHAO Jinliang, YIN Shaowu, GU Zhimin. Cloning and bioinformatics analysis of GH gene and its flanking region in Odontobutis potamophila [J]. , 2019, 31(9): 1461-1470.
[9]	YUAN Xiaolong, LI Yunqin, WANG Yi. Cloning and expression of three chalcone synthase-like genes from Paeonia delavayi [J]. , 2019, 31(9): 1478-1484.
[10]	PENG Shuai, CHEN Lang, ZHENG Tianyu, LU Huining, ZHANG Li, LIU Lixia. SNP site screening and bioinformatics analysis of TLR2 gene in Datong yak [J]. , 2019, 31(8): 1280-1285.
[11]	LIU Zhengkui, WU Yuan, CHEN Lin, WANG Lei, MU Hongye, ZHU Xuhang, WANG Xiaodu. Prokaryotic expression and bioinformatics analysis of Nsp5 gene of porcine epidemic diarrhea virus [J]. , 2019, 31(4): 532-538.
[12]	DONG Xinxing, LI Mingli, CUI Yijia, LAN Guoxiang, WANG Xiaoyi, YAN Dawei. Cloning, bioinformatics analysis and tissue expression detection of MCUR1 gene in Saba pig [J]. , 2019, 31(11): 1825-1833.
[13]	RUAN Xianle, WANG Junsheng, LIU Hongzhan, CHEN Liangbing, ZHAO Jinhui. Bioinformatics analysis of miR169 gene family in Brassica napus L. and prediction of their target genes [J]. , 2018, 30(8): 1273-1280.
[14]	NONG Weilun, BI Yingjie, LI Hongxu, ZHANG Li. H-FABP gene cloning, SNPs screening and bioinformatics analysis in Gansu black pig [J]. , 2018, 30(3): 378-385.
[15]	REN Weihe, REN Wenwen, MA Boyan, GAO Chenghong, LI Hongxu, LIU Lixia, CAO Xin, ZHANG Li. SNP screening and bioinformatics analysis of MSTN in Japanese White Rabbit [J]. , 2018, 30(12): 2018-2023.