鸡传染性支气管炎病毒S1基因抗原区的原核表达及鉴定

Abstract

Abstract: Avian infectious bronchitis virus (IBV) S1 gene was the object in this study and a pair of primers was designed for amplifying the S1 main antigen fragment of IBV, which was composed of 198 bp. And then it was sub-cloned into prokaryotic expression plasmid pGEX-6P-1. The recombinant plasmid pGEX-6P-1-S1 was transformed into E.coli Rosetta (DE3) and induced by IPTG. SDS-PAGE analysis showed a specific protein band with a molecular weight 33.0 ku, according to our study anticipation. The express production was inclusion body when it was identified. Western blotting indicated that the expressed protein could react with the chicken positive serum and be recognized by the specific monoclonal antibody of GST. The result suggested that the recombinant protein was correctly expressed and had a good antigenicity, which provided substantial antigen base for preparation of IBV monoclonal antibody.

Key words: avian infectious bronchitis virus; S1 protein; prokaryotic expression; identification

ZHU Yan-ping, GUO Dong-guang, YUE Feng, YANG Yuan, LI Chong, WANG Ai-guo, LI Bo-wen, RUAN Tao, KONG Ling-yun, WANG Xuan-nian. Prokaryotic Expression and Identification of the S1 Gene Main Antigen Domain of Avian Infectious Bronchitis Virus[J]. .

References

1 余娟，刘兴友，王玲丽. 鸡传染性支气管炎病毒致病机理的研究进展［J］. 中国畜牧兽医，2011，38(12)：160～162. 2 陈明艳，余娟，潘耀谦，等. 鸡传染性支气管炎病毒变异机制及致弱方法研究进展［J］.动物医学进展，2013，34(1)：1～5. 3 韩耘，李洪涛，姜力，等. 鸡传染性支气管炎病毒的分离鉴定［J］. 中国畜牧兽医，2011，38(3)：192～195. 4 Abro S H, Renstrm L H, Ullman K, et al. Characterization and analysis of the full-length genome of a strain of the European QX-like genotype of infectious bronchitis virus［J］. Archives of Virology, 2012, 157(6): 1211～1215. 5 Cavanagh D, Mawditt K, Welchman D B, et al. Coronaviruses from pheasants (Phasianus colchicus) are genetically closely related to coronaviruses of domestic fowl (infectious bronchitis virus) and turkeys［J］. Avian pathology, 2002, 31(1): 81～93. 6 de Wit J J, Cook J K A, Van der Heijden H M J F. Infectious bronchitis virus variants: A review of the history, current situation and control measures［J］. Avian Pathology, 2011, 40(3): 223～235. 7 Gelb J Jr, Ladman B S, Pope C R, et al. Characterization of nephropathogenic infectious bronchitis virus DMV/1639/11 recovered from Delmarva broiler chickens in 2011［J］. Avian Diseases Research, 2013, 51(7): 65～70. 8 Kim B Y, Lee D H, Jang J H, et al. Cross-protective immune responses elicited by a Korean variant of infectious bronchitis virus［J］. Avian Diseases, 2013, 57(4): 667～670.〖HJ〗 9 McKinley E T, Jackwood M W, Hilt D A, et al. Attenuated live vaccine usage affects accurate measures of virus diversity and mutation rates in avian coronavirus infectious bronchitis virus［J］. Virus Research, 2011, 158(1～2): 225～234. 10 Thor S W, Hilt D A, Kissinger J C, et al. Recombination in avian gamma-coronavirus infectious bronchitis virus［J］. Viruses, 2011, 3(9): 1777～1799. 11 Toffan A, Terregino C, Mazzacan E, et al. Detection of Chinese Q1 strain of infectious bronchitis virus in Europe［J］. Veterinary Research, 2011, 169(8): 1212～1213. 12 Xie Q, Ji J, Xie J, et al. Epidemiology and immunoprotection of nephropathogenic avian infectious bronchitis virus in southern China［J］. Virology Journal, 2011, 27(8): 484～488. 13 Yan F, Zhao Y, Yue W, et al. Phylogenetic analysis of S1 gene of infectious bronchitis virus isolates from China［J］. Avain Diseases, 2011, 55(3): 451～458. 14 Yang J T, Ma B C. Complete genome sequence of a nephropathogenic infectious bronchitis virus strain isolated in China［J］. Genome Announcements, 2013, 1(5): 1～2.

[1]	JIAO Linlin, CHENG Yuting, WU Qingguo, WU Shuang, WU Zhi, ZHU Shanyuan, QIAN Yingjuan. Prokaryotic Expression and Polyclonal Antibody Preparation of Duck Tembusu Virus Capsid Protein [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2395-2402.
[2]	LI Can, LI Chuchu, ZENG Wei, ZHANG Ning, JI Chunxiao, CHEN Tao. Recombinant Expression and Function Study of RegⅢγ Protein in Porcine [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2414-2426.
[3]	SONG Yue, SU Shengjie, ZHANG Fan, BAI Fan, DAI Lingli, WANG Na, WANG Dawei, YANG Qianwen, ZHAO Shihua, ZHANG Yuemei. Prokaryotic Expression and Polyclonal Antibody Preparation of Mannheimia haemolytica Truncated Leukotoxin [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2479-2486.
[4]	ZHANG Fangyuan, LIN Yating, YANG Dawei, CHEN Hu, LI Guimei, SHAN Hu. Prokaryotic Expression of African Swine Fever Virus P30 Protein and Establishment of Indirect ELISA Detection Method [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2012-2022.
[5]	HU Leyu, HU Xinyao, LI Ying, ZHAO Shengmiao, SHEN Fengzhen, WANG Changfa, LI Liangliang, WANG Tongtong, REN Huiying. Bioinformatics Analysis of HO-1 Gene and Preparation of Polyclonal Antibody in Dezhou Donkey [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1499-1510.
[6]	WANG Wenjie, RU Penghui, YU Chuan, DU Fuxi, CHEN Songbiao, SHANG Ke, ZHANG Chunjie, CHENG Xiangchao. Prokaryotic Expression of VP2 Protein of Luoyang Feline Panleukopenia Virus Isolate and Polyclonal Antibody Preparation [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1511-1521.
[7]	LIU Jiaxing, HAN Xueying, ZHANG Xinru, DENG Jiaxin, YAO Lunguang, LIU Yangkun. Preparation and Immunogenicity Analysis of Ferritin Nanoparticles Carrying Antigenic Epitopes of Porcine Epidemic Diarrhea Virus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1567-1574.
[8]	JIA Xinyue, MA Jing, ZHANG Yanyan, MA Xun, WANG Zhengrong, BO Xinwen. Bioinformatics Analysis and Prokaryotic Expression of EGR-07243 and EGR-07243 Genes of Echinococcus granulosus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 847-858.
[9]	ZHANG Zhitao, LI Xianglong, KONG Lingli, LUO Yuxin, WANG Dongying. Prokaryotic Expression of Serine/Threonine Protein Phosphatase 2A in Fasciola gigantica and Expression Analysis at Different Stages [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1107-1117.
[10]	GAN Lu, ZHENG Huizhen, NUO Mingdalai, LIU Yan, JIN Min, HE Wenwen, WEN Licui, LI Yongchang, GAILIKE·Bayinchahan. Propagation of Trypanosoma evansi Yili Strain and Cloning, Expression and Bioinformatics Analysis of Its PFR Gene [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 469-478.
[11]	LYU Shuangfei, MA Xun, WANG Jing, KONG Cuilian, KOU Lijun, LIU Caixia, SHI Weidi, KANG Lichao, REN Huijie. Prokaryotic Expression and Immunoreactivity of PA-LF1 Fusion Protein of Bacillus anthracis [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 674-683.
[12]	ZHANG Kai, ZHANG Chunping, GE Shengqiang, SUN Chunxi, CHENG Jie, FU Chunyu, WANG Gang, NIU Xing, PENG Jun. Expression of p22 Protein of African Swine Fever Virus,Preparation of Polyclonal Antibody and Establishment of Indirect ELISA Method [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 270-279.
[13]	ZHU Lilin, WANG Houkun, CHEN Xueyang, LIU Jing, LIANG Xiongyan, FANG Chun, YANG Yuying. Effect of Chicken IFIT5 on Replication of J Subgroup Avian Leukemia Virus Within DF-1 Cells [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 310-316.
[14]	WANG Yuchen, TIAN Guangyuan, ZHOU Yaping, GUO Ting, ZHAO Hongmei, SUN Yajie, ZHAO Fengmiao, BIAN Yuchen, YU Jialiang, HAO Yongqing. Prokaryotic Expression of BPIV3 NP Protein and Its Immunoenhancing Effect on the BPIV3 Inactivated Vaccine [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 3122-3130.
[15]	WANG Wei, HAN Huihui, DING Naizheng, HE Chengqiang. Prokaryotic Expression of Bovine Viral Diarrhea Virus E0 Protein and Preparation of Polyclonal Antibody [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 3131-3139.

Prokaryotic Expression and Identification of the S1 Gene Main Antigen Domain of Avian Infectious Bronchitis Virus

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments