猪流行性腹泻病毒部分N基因的原核表达及表达产物反应原性分析

doi:10.16431/j.cnki.1671-7236.2016.03.011

Abstract

Abstract: This experiment was aimed to study the antigenicity of prokaryotic expression of the N gene fragment of porcine epidemic diarrhea virus (PEDV),and lay a foundation for establishing an indirect ELISA method of PEDV.The N gene segment was amplified by RT-PCR,then the recombinant plasmid with the vector pET-30a(+) was constructed,which was induced by 0.5 mmol/L IPTG at 37 ℃ for 4 h.Furthermore,the expressed product was analyzed by SDS-PAGE and Western blotting.Sequencing results proved that recombinant plasmid was correctly constructed.The result showed that the PEDV polyclonal antibody could specifically bind to PEDV N protein,which indicated that the recombinant fusion protein had excellent immunogenicity.A prokaryotic expression vector for the fragment of N protein was successfully constructed in this study,which laid a foundation for the development of diagnosis of PEDV.

Key words: porcine epidemic diarrhea virus (PEDV); N gene fragment; prokaryotic expression; reactionogenicity

CLC Number:

S852.65

ZHANG Bo, LI Shou-jun, LV Mao-jie, YANG Bao-shou. Prokaryotic Expression of Partial N Gene Fragment of Porcine Epidemic Diarrhea Virus and Reactionogenicity Analysis of the Expressed Protein[J]. , 2016, 43(3): 637-643.

References

[1] Pensaert M B,Debouck P.A new coronavirus-like particle associated with diarrhea in swine[J].Arch Virol,l978,58(3):243-247.
[2] Kocherhans R,Bridgen A,Ackermann M,et al.Completion of the porcine epidemic diarrhoea coronavirus (PEDV) genome sequence[J].Virus Genes,2001,23(2):137-144.
[3] Park S J,Kim H K,Song D S,et al.Complete genome sequences of a Korean virulent porcine epidemic diarrhea virus and its attenuated counterpart[J].J Virol,2012,86(10):5964.
[4] Pijpers A,van Nieuwstadt A P,Terpstra C,et al.Porcine epidemic diarrhoea virus as a cause of persistent diarrhoea in a herd of breeding and finishing pigs[J].Vet Rec,1993,132(6):129-131.
[5] Carvajal A,Diego R,Lanza I,et al.Seroprevalence of porcine epidemic diarrhea virus infection among difference types of breeding swine farms in Spain[J].Preventive Veterinary Medicine,1995,23(1):33-40.
[6] Song D,Park B.Porcine epidemic diarrhoea virus:A comprehensive review of molecular epidemiology,diagnosis,and vaccines[J].Virus Genes,2012,44(2):167-175.
[7] Lee H K,Yeo S G.Cloning and sequence analysis of the nucleocapsid gene of porcine epidemic diarrhea virus Chinju99[J].Virus Genes,2003,26(2):207-212.
[8] Wood E N.An apparently new syndrome of porcine epidemic diarrhoea[J].Vet Rec,1977,100(12):243-244.
[9] Sueyoshi M,Tsuda T,Yamazaki K,et al.An immunohistochemical investigation of porcine epidemic diarrhea[J].J Comp Pathol,1995,113(1):59-67.
[10] Hwang E K,Yoon K J,Jean Y H,et al.Current occurrence of porcine epidemic diarrhoea in Korea[J].Rda Journal of Agricultural Science,1994,36(2):587-596.
[11] Chae C,Kim O,Choi C,et al.Prevalence of porcine epidemic diarrhoea virus and transmissible gastroenteritis virus infection in Korean pigs[J].Vet Rec,2000,147(21):606-608.
[12] 朱迪国,宋建德,袁丽萍,等.2013—2014年全球猪流行性腹泻重大疫情分析[J].中国动物检疫,2014,31(10):42-46.
[13] 王金良,谢金文,唐娜,等.应用逆转录套式PCR检测猪流行性腹泻病毒[J].中国畜牧兽医,2012,39(9):86-88.
[14] 刘云波,赵洪翠,王志成,等.猪病毒性腹泻分子流行病学调查[J].中国畜牧兽医,2013,40(2):204-207.
[15] 陈申秒,牛成明,何福庆,等.猪流行性腹泻病毒研究进展及疫苗应用前景[J].中国畜牧兽医,2014,41(3):223-229.
[16] 石达,陈建飞,时红艳,等.猪流行性腹泻病毒N蛋白与Vero E6 核仁蛋白Fibrillarin的亚细胞定位分析[J].中国预防兽医学报,2011,33(11):833-836.
[17] 张志榜,陈建飞,时洪艳,等.猪流行性腹泻病毒CH/S株M蛋白全长的原核表达及其反应原性分析[J].中国兽医科学,2011,41(1):31-35.

Prokaryotic Expression of Partial N Gene Fragment of Porcine Epidemic Diarrhea Virus and Reactionogenicity Analysis of the Expressed Protein

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[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]	HUO Mingkai, GUAN Feihu, QIU Runhui, WEI Chunyan, YU Haitao, ZHU Jiale, ZHANG Hui. Study on the Mechanism of DDX1 Inhibiting Porcine Epidemic Diarrhea Virus Replication [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1556-1566.
[8]	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.
[9]	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.
[10]	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.
[11]	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.
[12]	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.
[13]	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.
[14]	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.
[15]	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.