截短猪流行性腹泻病毒N蛋白的可溶性表达及其抗原活性

doi:10.11843/j.issn.0366-6964.2014.09.026

Abstract

Abstract:

To express the porcine epidemic diarrhea virus (PEDV) N protein by prokaryotic expression system and to identify its antigenicity，truncated N protein (tN) genes were amplified by RT-PCR and recombinant prokaryotic expression plasmid (pET32a-PEDV-tN) with tN protein gene was constructed.After the recombinant plasmids had been transferred into competent cells of E.coli BL21 and the bacteria had been inducted by IPTG，the expression and immunoreactivity of tN protein were detected respectively by SDS-PAGE and Western blotting.Then，BLAB/c mice were subcutaneously immunized with purified tN protein and the protein immunogenicity was detected by ELSIA and Immunoperoxidase monolayer assay.The tN protein genes of 705 bp were amplified and the recombinant plasmids carrying this gene could solubly express recombinant tN protein under induction of IPTG in E.coli BL21.The protein displayed specific binding to swine anti-PEDV antibody and induced mice to produce anti-PEDV antibody.These results showed that the soluble expression of PEDV tN protein with favorable complete antigenic activities was implemented by prokaryotic expression system.

CLC Number:

S852.659.6

ZHU Wei-xia，YUAN Wan-zhe，LI Li-min，SONG Qin-ye，SUN Tai-ran，LI Tan-qing. Prokaryotic Soluble Expression of Trunked N Protein of Porcine Epidemic Diarrhea Virus and Its Antigenic Activity[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(9): 1561-1566.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.xmsyxb.com/EN/10.11843/j.issn.0366-6964.2014.09.026

https://www.xmsyxb.com/EN/Y2014/V45/I9/1561

References

［1］PAN Y，TIAN X，LI W，et al.Isolation and characterization of a variant porcine epidemic diarrhea virus in China［J］.Virol J,2012，9:195.doi：10.1186/1743-422X-9-195.
［2］LI W，LI H，LIU Y，et al.New variants of porcine epidemic diarrhea virus，China，2011［J］.Emerg Infect Dis，2012，18(8):1350-1353.
［3］PURANVEJA S，POOLPERM P，LERTWATCHARASASAKUL P，et al .Chinese-like strain of porcine epidemic diarrhea virus，Thailand ［J］.Emerg Infect Dis，2009，15(7)：1112-1115.
［4］OLANRATMANEE E O，KUNAVONGKRIT A，TUMMARUK P.Impact of porcine epidemic diarrhea virus infection at different periods of pregnancy on subsequent reproductive performance in gilts and sows［J］.Anim Reprod Sci，2010，122(1-2):42-51.
［5］ZHANG Q，HU R，TANG X，et al.Occurrence and investigation of enteric viral infections in pigs with diarrhea in China［J］.Arch Virol，2013，158(8):1631-1636.
［6］KUBOTA S，SASAKI O，AMIMOTO K，et al.Detection of porcine epidemic diarrhea virus using polymerase chain reaction and comparison of the nucleocapsid protein genes among strains of the virus［J］.J Vet Med Sci，1999，61(7):827-830.
［7］PARK S J，SONG D S，PARK B K.Molecular epidemiology and phylogenetic analysis of porcine epidemic diarrhea virus (PEDV) field isolates in Korea［J］.Arch Virol,2013，158(7):1533-1541.
［8］PURANAVEJA S，POOLPERM P，LERTWATCHARASARAKUL P，et al.Chinese-like strain of porcine epidemic diarrhea virus，Thailand［J］.Emerg Infect Dis,2009，15(7):1112-1115.
［9］HOANG H，KILLIAN M L，MADSON D M，et al.Full-length genome sequence of a plaque-cloned virulent porcine epidemic diarrhea virus isolate (USA/Iowa/18984/2013) from a Midwestern U.S.swine herd［J］.Genome Announc，2013，1(6).pii：e01049-13.
［10］HUANG Y W，DICKERMAN A W，PIEYRO P，et al.Origin，evolution，and genotyping of emergent porcine epidemic diarrhea virus strains in the United States［J］.mBio，2013，4(5):e00737-13.
［11］BRIAN D A，BARIC R S.Coronavirus genome structure and replication［J］.Curr Top Microbiol Immunol，2005，287:1-30.
［12］DUARTE M，GELFI J，LAMBERT P，et al.Genome organization of porcine epidemic diarrhoea virus［J］.Adv Exp Med Biol，1993，342:55-60.
［13］TOBLER K，BRIDGEN A，ACKERMANN M.Sequence analysis of the nucleocapsid protein gene of porcine epidemic diarrhoea virus［J］.Adv Exp Med Biol，1993，342：49-54.
［14］LIU D Q，GE J W，QIAO X Y，et al.High-level mucosal and systemic immune responses induced by oral administration with Lactobacillus-expressed porcine epidemic diarrhea virus (PEDV) S1 region combined with Lactobacillus-expressed N protein ［J］.Appl Microbiol Biotechnol，2012，93(6):2437-2446.
［15］杨巍，李广兴，孟凡丹，等.猪流行性腹泻病毒N基因的克隆及原核表达［J］.东北农业大学学报，2011，42（6）：81-85.
［16］LADEKJAER-MIKKELSEN A S，NIELSEN J，STADEJEK T，et al.Reproduction of postweaning multisystemic wasting syndrome (PMWS) in immunostimulated and non-immunostimulated 3-week-old piglets experimentally infected with porcine circovirus type 2 (PCV2) ［J］.Vet Microbiol,2002，89(2-3)：97-114.

[1]	SONG Wenyan, ZHANG Hanwen, WU Aodi, ZHANG Liyan, LIU Zhao, YE Tongtong, CHEN Chuangfu, SHENG Jinliang. Screening of Nanobodies against Porcine Reproductive and Respiratory Syndrome Virus GP5 Protein and Exploration of Their Inhibitory Effect on Virus Replication [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 258-270.
[2]	WANG Ying, LI Jiakang, ZENG Yue, SHI Kaituo, WEI Liting, PENG Jiajia, LI Qiuyan, CAO Longlong, ZHOU Dengyuan. Development of Monoclonal Antibodies against Feline Calicivirus [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 401-405.
[3]	LI Huihui, DONG Keer, LIU Xinbo, ZHANG Chunxiao, MA Chao, CHEN Liping, ZHONG Qi, YAO Gang, MA Xuelian. Establishment and Application of Rapid Detection Method for Bovine Norovirus and Bovine Rotavirus Dual RAA-LFD [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 406-412.
[4]	ZHOU Jianhao, WANG Dongfang, LIU Ying, WANG Shujuan, MA Zhenyuan, XIE Caihua, ZHAO Xueli, YANG Haibo, FENG Guidan, KANG Taisheng, HU Yufeng, LI Bowen, YAN Ruoqian. Establishment and Preliminary Application of a Quantitative Droplet Digital PCR Assay for Porcine Epidemic Diarrhea Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 413-418.
[5]	ZHANG Jiaqi, JIA Xining, ZHOU Qun, SONG Xin, ZHU Chenxi, LI Gege, ZHANG Bin. Construction of BVDV-1 Virus-like Particles based on E2 Protein and Evaluation of Its Immune Efficacy in Mice [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 5143-5153.
[6]	ZHOU Likun, DONG Xinyuan, LI Jiahui, CUI Zhiying, ZHAO Shijie, XU Jie, CHEN Jing, ZHANG Yina, XIA Ping’an. Regulatory Role of MID2 on Porcine Reproductive and Respiratory Syndrome Virus Replication [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4735-4744.
[7]	ZHENG Ruwen, HUANG Tao, WU Daoyi, YU Guangmei, LI Fangfang, SUI Xin, MIN Tingyu. Establishment and Preliminary Application of LAMP-LFD Detection Method for Bovine Viral Diarrhea Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4745-4753.
[8]	JIA Yiquan, YU Mengting, LIU Weirong, FANG Shouguo, ZHANG Songbai. Baicalin Inhibits Infectious Bronchitis Virus Replication via Promoting the Expression of Type Ⅰ Interferon [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4839-4850.
[9]	CHEN Lu, CHANG Xinyu, SHEN Jiachen, SHEN Ruiting, ZHAO Zhenhua, HOU Xiaolin. Transcriptome Sequencing and Analysis of HD11 Cells Infected by Avian Infectious Bronchitis Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4860-4865.
[10]	HE Yeqing, ZUO Baijiao, LI Qinghao, SUN Juan, JIN Xin, XIA Lu, WEI Zhanyong. Histopathological Observation of Immune Organs of Piglets Infected with Transmissible Gastroenteritis Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4872-4879.
[11]	CHEN Wenzhe, ZHANG Xiangle, GU Fengxing, ZHAO Zhenxiang, LI Kangli, XUE Zhaoning, ZHENG Haixue, ZHANG Xiaoli, ZHU Zixiang. Evaluation of the Antiviral Effect of Porcine GRK2 against Foot-and-mouth Disease Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4350-4361.
[12]	ZOU Hong, XIA Yingju, LI Ling, XU Lu, ZHAO Junjie, WANG Tuanjie, ZHANG Qianyi, SONG Zhenhui. Establishment and Preliminary Application of a Dual Real-time RT-PCR Assay for CSFV and BVDV [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4422-4427.
[13]	YUAN Li, SUN Yangyang, ZHANG Lujie, ZHANG Jie, SUN Haifeng, BAI Juan, JIANG Ping. Production and Epitope Characterization of Monoclonal Antibodies against GP3 Protein of Porcine Reproductive and Respiratory Syndrome Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3424-3434.
[14]	CAO Liyan, KONG Xiangyu, LI Xiangtong, SUO Xuepeng, DUAN Yueyue, YUAN Cong, SHI Lei, ZHANG Yu, MA Guoxiang, ZHENG Haixue, WANG Qi. Preparation and Sequence Analysis of Monoclonal Antibody against the Nucleocapsid Protein of Porcine Acute Diarrhea Syndrome Coronavirus [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2487-2497.
[15]	ZHANG Senhao, WANG Xueying, CAI Limeng, XIE Weichun, KUANG Hongdi, WANG Xiaona, LI Jiaxuan, CUI Wen, JIANG Yanping, ZHOU Han, SHAN Zhifu, WANG Li, QIAO Xinyuan, LI Yijing, TANG Lijie. Establishment and Application of Rapid Diagnosis Method for RPA of Porcine Epidemic Diarrhea Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2498-2508.

Prokaryotic Soluble Expression of Trunked N Protein of Porcine Epidemic Diarrhea Virus and Its Antigenic Activity

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments