猪乙型脑炎病毒E蛋白的原核表达与鉴定

Abstract

Abstract: A fragment of about 972 bp long was amplified by RT-PCR technique with a pair of specific primers based on published Japanese encephalitis virus(JEV) genome sequence. Then the target fragment was directionally cloned into pET30a vector. After identifying with enzyme cutting and sequencing, the recombinant plasmid was transformed into E.coli BL21(DE3). The recombinant protein E was expressed in inclusion body form in E.coli after induction with IPTG. After denaturation, purification and renaturation, the purified protein was analyzed by Western blotting, the results showed that the purified recombinant protein retained better antigenicity and specificity.

Key words: Japanese encephalitis virus; E protein; prokaryotic expression; activity detection

CLC Number:

S852.65

CHEN Guo-qiang, DIAO Fu-hua. Expression and Identification of Japanese Encephalitis Virus Truncated E Protein in E.coli[J]. , 2012, 39(10): 80-82.

References

1 丛晓燕,吴家强,王怀忠,等.猪乙型脑炎病毒E基因的克隆与序列分析[J].中国畜牧兽医,2009,36(9):71~73.
2 冯若飞, 李向茸, 韦鹏建, 等. 乙型脑炎病毒E基因Ⅰ、Ⅱ抗原域片段的克隆与原核表达[J]. 西北民族大学学报(自然科学版), 2011,32(1):68~72.
3 陈娜莎, 华荣虹, 闫丽萍, 等. 乙型脑炎病毒E蛋白抗原表位多肽序列鉴定及分析[J]. 中国预防兽医学报, 2011,33(7):541~545.
4 谢荣辉, 朱函坪, 张晓锋, 等. 浙江省猪血清中乙脑病毒的分离鉴定及抗体的测定[J]. 中国人兽共患病学报, 2010,26(12):1123~1125.
5 蔡绪禹, 康晓平, 刘洪, 等. 5种脑炎人兽共患病病毒多重RT-PCR检测方法的建立[J]. 中国预防兽医学报, 2011,33(2):118~121.
6 Chen H W, Pan C H, Liau M Y, et al. Screening of protective antigens of Japanese encephalitis virus by DNA immunization:A comparative study with conventional viral vaccines[J]. J Virol,1999,73(12):10127~10145.
7 Chen W R, Tesh R B, Rico-Hesse R. Genetic variation of Japanese encephalitis virus in nature[J]. J Gen Virol,1990,71(12):2915~2922.
8 Kolaskar A S, Kulkarni-Kale U. Prediction of three-dimensional structure and mapping of conformational epitopes of envelope glycoprotein of Japanese encephalitis virus[J]. Virology,1999, 261(1):31~42.
9 Mandl C W, Guirakhoo F, Holzmann H, et al. Antigenic structure of the flavivirus envelope protein E at the molecular level, using tick-borne encephalitis virus as a model[J]. J Virol,1989, 63(2):564~571.
10 McMinn P C. The molecular basis of virulence of the encephalitogenic flaviviruses[J]. J Gen Virol,1997,78(11):2711~2722.
11 Takasaki T. Japanese encephalitis virus and other flaviviruses[J]. Nippon Rinsho,2010,68(6):363~366.
12 Tang W F, Ogawa M, Eshita Y, et al. Molecular evolution of Japanese encephalitis virus isolates from swine in Oita, Japan during 1980—2009[J]. Infect Genet Evol,2010,10(2):329~336.
13 Yang Y, Ye J, Yang X H, et al. Japanese encephalitis virus infection induces changes of mRNA profile of mouse spleen and brain[J]. Virol J, 2011,8:80.

[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.

Expression and Identification of Japanese Encephalitis Virus Truncated E Protein in E.coli

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments