[1]赵冬敏,刘宇卓,黄欣梅,等.坦布苏病毒E蛋白结构域I/II蛋白免疫保护效果[J].江苏农业学报,2017,(02):379-383.[doi:doi:10.3969/j.issn.1000-4440.2017.02.022]
 ZHAO Dong-min,LIU Yu-zhuo,HUANG Xin-mei,et al.Immunoprotective effect of Tembusu virus E protein domains I and II[J].,2017,(02):379-383.[doi:doi:10.3969/j.issn.1000-4440.2017.02.022]
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坦布苏病毒E蛋白结构域I/II蛋白免疫保护效果()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2017年02期
页码:
379-383
栏目:
畜牧兽医·水产养殖
出版日期:
2017-04-30

文章信息/Info

Title:
Immunoprotective effect of Tembusu virus E protein domains I and II
作者:
赵冬敏刘宇卓黄欣梅韩凯凯杨婧刘青涛毕可然李银
(江苏省农业科学院兽医研究所/国家兽用生物制品工程技术研究中心/农业部兽用生物制品工程技术重点实验室,江苏南京210014)
Author(s):
ZHAO Dong-minLIU Yu-zhuoHUANG Xin-meiHAN Kai-kaiYANG JingLIU Qing-taoBI Ke-ranLI Yin
(Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences/Key Laboratory of Veterinary Biologicals Engineering and Technology, Ministry of Agriculture/National Center for Engineering Research of Veterinary Bio-products, Nanjing 210014, China)
关键词:
坦布苏病毒E蛋白结构域I/II免疫保护
Keywords:
Tembusu virusE proteindomains I and IIimmunoprotection
分类号:
S852.65+9.6
DOI:
doi:10.3969/j.issn.1000-4440.2017.02.022
文献标志码:
A
摘要:
为了探讨原核表达的坦布苏病毒E蛋白结构域I/II蛋白对于坦布苏病毒感染的免疫保护作用,利用IPTG诱导重组大肠杆菌表达坦布苏病毒E蛋白结构域I/II蛋白,Anti-FLAG M2 Affinity Gel纯化后进行SDS-PAGE和Western blot鉴定。经SDS-PAGE和Western blot鉴定,纯化后坦布苏病毒E蛋白结构域I/II蛋白仅有单一目的条带。免疫组小鼠血清ELISA效价均达 4.67±0.11 (lg10)。攻毒保护试验结果表明,免疫组小鼠体质量在攻毒9 d后高于对照组,荧光定量RT-PCR检测结果显示,攻毒后免疫组小鼠脑、肝脏、脾脏组织中坦布苏病毒核酸含量均低于对照组。表明使用原核表达的坦布苏病毒E蛋白结构域I/II蛋白免疫可诱导产生高水平抗体,并提供良好的免疫保护力,为开发有效的坦布苏病毒亚单位疫苗提供依据。
Abstract:
The immunogenicity of the prokaryotically-expressed Tembusu virus E protein domains I and II was tested by SDS-PAGE and Western blot identification of Tembusu virus E protein domains I and II expressed in Escherichia coli and induced by IPTG. The results showed that only a single target band was detected after purification. The titers of antibody of immunized group reached 4.67±0.11(lg10). The weight of immunized group was higher than that of control group from the 9th day postinoculation. Real-time RT-PCR showed that the Tembusu virus nucleic acid in the brain, liver and spleen of mice in immunized group was less than that of control group. It was indicated that the immunization with recombinant tembusu virus E protein domains I and II could induce high level of specific antibody and provide immuno protection against Tembusu virus infection.

参考文献/References:

[1]BAI X, LV R, LIU C, et al. Molecular characterization of a duck tembusu virus from China[J]. Virus Genes, 2013, 47(3):478-482.
[2]YOUN S, CHO H, FREMONT D H, et al. A short N-terminal peptide motif on flavivirus nonstructural protein NS1 modulates cellular targeting and immune recognition[J]. J Virol, 2010, 84 (18): 9516-9532.
[3]KOLASKAR A S, URMILA K K. Prediction of three-dimensional structure and mapping of conformational epitopes of envelope glycoprotein of Japanese encephalitis virus[J]. Virology, 1999, 261(1):31-42.
[4]MUKHOPADHYAY S, KUHN R J. A structural perspective of the flavivirus life cycle[J]. Nat Rev Microbiol, 2005, 3(1):13-22.
[5]HURRELBRINK R J, MCMINN P C. Molecular determinants of virulence:the structural and functional basis for flavivirus attenuation[J]. Adv Virus Res, 2003, 60(1):1-42.
[6]MENDES Y S, ALVES N S, SOUZA T L, et al. The structural dynamics of the flavivirus fusion peptide-membrane interaction[J]. PLoS One, 2012, 7(10):e47596.
[7]YU K, SHENG Z Z, HUANG B, et al. Structural, antigenic, and evolutionary characterizations of the envelope protein of newly emerging duck tembusuvirus[J]. PLoS One, 2013, 8(8):e71319.
[8]赵冬敏,黄欣梅,刘宇卓,等. 鹅坦布苏病毒E蛋白质结构域I和II在大肠杆菌中的表达及鉴定[J]. 江苏农业学报, 2014, 30(3): 590-594.
[9]LIU M, CHEN S, CHEN Y, et al. Adapted tembusu-like virus in chickens and geese in China[J]. J Clin Microbiol, 2012, 50(8):2807-2809.
[10]HUANG X, HAN K, ZHAO D, et al. Identification and molecular characterization of a novel flavivirus isolated from geese in China[J]. Res Vet Sci, 2013, 94(3): 774-780.
[11]TANG Y, GAO X, DIAO Y, et al. Tembusu virus in human, China[J]. Transbound Emerg Dis, 2013, 60(3):193-196.
[12]LEDIZET M, KAR K, FOELLMER HG, et al. A recombinant envelope protein vaccine against West Nile virus[J]. Vaccine, 2005, 23(30):3915-3924.
[13]MUTOH E, ISHIKAWA T, TAKAMIZAWA A, et al. Japanese encephalitis subunit vaccine composed of virus-like envelop antigen particles purified from serum-free medium of a high-producer J12#26 cell clone[J]. Vaccine, 2004, 22(20):2599-2608.

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备注/Memo

备注/Memo:
收稿日期:2016-06-22 基金项目:江苏省自然科学基金项目(BK20160064);江苏省农业科技自主创新基金项目[CX(14)2091] 作者简介:赵冬敏(1982-),女,山东兖州人,博士,副研究员,主要从事家禽重大疫病流行病学及致病分子机制研究工作。(E-mail)zhaodongmin126@126.com 通讯作者:李银,(E-mail)muziyin08@163.com
更新日期/Last Update: 2017-05-02