伪狂犬病病毒gB蛋白抗原表位基因串联构建及原核表达

doi:10.16431/j.cnki.1671-7236.2015.04.006

Abstract

Abstract: The two dominant epitope sequences of pseudorabies virus (PRV) gB gene were joined together by overlap-extension PCR method with two pairs of primers. The products were inserted into pMD18-T vector. After properly sequenced, the multi-epitope sequence was linked with pET-32a(+) vector, and then transformed into BL21(DE3). By being induced with IPTG, the targeted protein was detected by SDS-PAGE and Western blotting. The results showed the fusion protein was successfully expressed with the size of 54 ku including 34 ku tandem protein and could be combined with monoclonal antibody against gB which showed that the expressed protein had good antigenicity.

Key words: pseudorabies virus (PRV); gB gene; epitope; tandem gene expression

CLC Number:

Q786

WANG Yu, JI Yi-kuan, CHENG Yi, XIANG Ke-yu, ZHANG Bao-shi, JU Chun-mei. Construction of Tandem Epitopes of gB Gene of Pseudorabies Virus and its Prokaryotic Expression[J]. , 2015, 42(4): 810-815.

References

[1] Mcinerbey J,Koou D.Economic analysis of alternative Aujeszky's disease control programmers[J].Veterinary Microbiology,1997,55(1):113-120.
[2] Daneil D,Anne-Marie T,Véronique B,et al.Effective protection of pigs against lethal pseudorabies virus infection after a single injection of low-dose Sindbis-derived plasmids encoding PrV gB,gC and gD glycoproteins[J]. Vaccine,2005,23(26):3483-3491.
[3] Nixdorf R,Klupp B G,Karger A,et al.Effects of truncation of the carboxy terminus of pseudorabies virus glycoprotein B on infectivity[J].Journal of Virology,2000,74(15):7137-7145.
[4] 殷震,刘景华.动物病毒学[M].第2版.北京:科学出版社,1997.
[5] 包新奇,黄绍华,刘巧荣,等.猪伪狂犬病毒gB基因在大肠杆菌中的分段表达[J].中国比较医学杂志,2012,3:12-16.
[6] 王大伟.抗伪狂犬病毒单克隆抗体的制备及gB基因编码蛋白抗原表位的表达[D].洛阳:河南科技大学,2011.
[7] Zaripov M M,Morenkov O S,Shmatchenko V V,et al.Immunological and functional characteristics of epitopes and regions of gB glycoprotein of Aujeszky's disease virus[J].Vopr Virusol,2001,46(2):41-45.
[8] 孙磊磊,程艺,梁梓森,等.伪狂犬病毒EP0基因的克隆及其在大肠杆菌中的表达研究[J].中国畜牧兽医,2012,39(6):57-60.
[9] 程艺,王雨,吉艺宽,等.伪狂犬病病毒立即早期180基因的克隆及序列分析[J].中国畜牧兽医,2014,41(1):15-18.
[10] 童光志,陈焕春.伪狂犬病流行现状及中国应采取的防制措施[J].中国兽医学报,1999,1:4-5.
[11] 杨庆芳,宁官保,李俊达.猪伪狂犬病病毒的分离鉴定[J].山西农业科学,2011,8:886-889.
[12] 彭金美,安同庆,赵鸿远,等.猪伪狂犬病病毒新流行株的分离鉴定及抗原差异性分析[J].中国预防兽医学报,2013,1:1-4.
[13] 童武,张青占,郑浩,等.免疫后发病仔猪中伪狂犬病毒的分离和鉴定[J].中国动物传染病学报, 2013,3:1-7.

[1]	ZHANG Guoqing, JIANG Yuhang, ZHANG Shuang, LI Letian, HAO Jiayi, CHEN Jing, ZOU Wancheng, LI Chang, LI Taiyuan, GAO Xu, JIN Ningyi. Preparation and Identification of PDCoV RBD Protein Peptide Antibody [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1575-1583.
[2]	ZHANG Jingxu, XU Yu, LIANG Haiying, ZENG Zhiyong, TANG Deyuan, WANG Bin, XU Songping, WAN Juan, ZHU Yang. Phylogenetic Evolution and Epitope Analysis of Pseudorabies Virus Guizhou Isolates and Vaccine Strains [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1093-1106.
[3]	ZHOU Wenfeng, ZHANG Yingjie, BAI Yihan, ZHANG Luhua, MAI Jinhui, WANG Naidong, YANG Yi, ZHAN Yang, WANG Dongliang. Complete Genome Sequence Analysis and Cap CTL Epitopes Prediction of Porcine Circovirus Type 2 [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 656-665.
[4]	XU Yaling, ZHANG Jihui, NIU Xi, LI Sheng, HUANG Shihui, RAN Xueqin, WANG Jiafu. Construction of Synthetic Peptide Vaccine of African Classical Swine Fever Virus P72 Protein and Evaluation of Its Immune Efficacy [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 3083-3090.
[5]	XIE Sihao, GOU Hongchao, BIAN Zhibiao, LI Bin, CAI Rujian, ZANG Yingan, LI Chunling. Effects of MLKL Gene Knockout on Replication of Pseudorabies Virus [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1934-1941.
[6]	LI Shinian, LIU Wanning, CHEN Yaping, WANG Jintao. Design of Multi Epitope Vaccine Against Swine Acute Diarrhea Syndrome Coronavirus S, M and E Proteins Based on Immunoinformatics Methods [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 1057-1066.
[7]	XING Jiayou, FAN Wenjie, WANG Yumin, LU Xiuxiang, SONG Yue, CHU Beibei, WANG Jiang, WANG Mengdi. Effect of Tamoxifen on the Proliferation of Porcine Pseudorabies Virus in vitro [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(11): 4503-4512.
[8]	MO Hongfang, JIE Kai, CHEN Xin, WEN Wei, LIU Wenqiang, CHEN Huanchun, QIAN Ping, LI Xiangmin. Preparation and Identification of Monoclonal Antibodies Against Porcine Seneca Valley Virus [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(1): 307-317.
[9]	YANG Mingzhu, CHEN Xiaochun, SONG Jiacheng, HOU Yaxin, LI Junping. Research Progress on Construction and Application of Recombinant Animal Herpesvirus Live Vector Vaccine [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(1): 338-351.
[10]	PAN Chenhao, ZHANG Xin, ZHAO Ruili, JIANG Xuan, JIN Tianming, YU Enyuan, LI Liuan, ZENG Jun, YU Xiaoxue, SONG Qiqi, HU Ye. Construction and Cytotoxicity Detection of Recombinant Epitope Protein DCpep-GSE of Streptococcus suis [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(8): 2990-3001.
[11]	QU Zhehui, ZHANG Xiwen, ZHAO Yu, ZHENG Quanfang, LIAN Huixiang, ZHAO Yunhuan, GUO Xiaoqiu. Genetic Variation Analysis of Major Virulence Genes of Pseudorabies Virus Epidemic Strains in Southern Henan Province [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(8): 3019-3029.
[12]	QU Zhehui, ZHAO Yu, ZHANG Xiwen, LIAN Huixiang, ZHAO Yunhuan, CHEN Bin, GUO Xiaoqiu. Expression of Pseudorabies Virus gE Protein and Preparation of Its Polyclonal Antibody [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(2): 631-639.
[13]	HE Xinglin, ZOU Zhong, GONG Wenxiao, ZHANG Yufei, LI Chengfei, XU Ting, CHEN Huanchun, JIN Meilin. Construction and Identification of Recombinant Pseudorabies Virus Expressing the p54 Protein of African Swine Fever Virus [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(11): 4182-4191.
[14]	ZHANG Li, REN Weike, LU Chao, LI Xiuli, LI Fuqiang, TIAN Xiangxue, CHI Jingjing, WANG Lili, JIANG Shan, DONG Zhimin, YAN Minghua. Genetic Variation Analysis of gB, gC and gE Genes of Porcine Pseudorabies Viruses Isolated in Tianjin During 2015-2020 [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(10): 3741-3751.
[15]	CHANG Wenru, DUAN Lifang, YANG Le, MA Yingxian, WANG Qi, ZHAI Yunyun, ZHOU Luyu, ZHANG Shuang, MING Shengli, YANG Guoyu, WANG Jiang, CHU Beibei. Study on Replication Patterns of Pseudorabies Virus in Cell Lines with NF-κB Family p65 Gene Knocked Out [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(1): 83-92.

Construction of Tandem Epitopes of gB Gene of Pseudorabies Virus and its Prokaryotic Expression

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments