[1]赵冬敏,黄欣梅,刘宇卓,等.鹅坦布苏病毒囊膜蛋白结构域II在大肠杆菌中的表达及免疫学鉴定[J].江苏农业学报,2015,(03):619-623.[doi:10.3969/j.issn.1000-4440.2015.03.025]
　ZHAO Dong-min,HUANG Xin-mei,LIU Yu-zhuo,et al.Expression and immunology of domain II of goose tembusu virus envelope protein in Escherichia coli[J].,2015,(03):619-623.[doi:10.3969/j.issn.1000-4440.2015.03.025]
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鹅坦布苏病毒囊膜蛋白结构域II在大肠杆菌中的表达及免疫学鉴定()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2015年03期
页码:
619-623
栏目:
畜牧兽医·水产养殖
出版日期:
2015-06-30

文章信息/Info

Title:
Expression and immunology of domain II of goose tembusu virus envelope protein in Escherichia coli
作者:
赵冬敏; 黄欣梅; 刘宇卓; 韩凯凯; 杨婧; 谢星星; 刘晓燕; 李银
（江苏省农业科学院兽医研究所，农业部兽用生物制品工程技术重点实验室，国家兽用生物制品工程技术研究中心，江苏南京210014）
Author(s):
ZHAO Dong-min; HUANG Xin-mei; LIU Yu-zhuo; HAN Kai-kai; YANG Jing; XIE Xing-xing; LIU Xiao-yan; LI 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:
鹅坦布苏病毒; 囊膜蛋白; 结构域II
Keywords:
goose tembusu virus; envelope protein; domain II
分类号:
S858.322.5+2
DOI:
10.3969/j.issn.1000-4440.2015.03.025
文献标志码:
A
摘要:
为了实现鹅坦布苏病毒囊膜蛋白（E蛋白）结构域II在大肠杆菌中的表达并对其进行免疫学鉴定，根据鹅坦布苏病毒JS804株囊膜蛋白基因序列，利用人工合成法获得鹅坦布苏病毒囊膜蛋白结构域II编码基因并引入限制性酶切位点，片段大小为507 bp。将合成的基因片段克隆入pGEX-4t-1原核表达载体中，构建重组表达载体pGEX-EII并转化至BL21（DE3）中。通过优化表达条件获得重组蛋白质并进行免疫学鉴定。结果显示，重组蛋白质分子量约为 4.4×104，Western-blot和间接免疫荧光检测结果表明重组蛋白质具有良好的免疫原性。
Abstract:
To express and immunoidentify goose tembusu virus envelope domain II in Escherichia coli, the encoding gene of goose tembusu virus JS804 strain envelope protein domain II was artificially synthesized with the length of 507 bp. The synthesized gene was then inserted into the prokaryotic vector pGEX-4t-1 for the construction of recombinant expression plasmid pGEX-EII and then transformed into E. coli BL21 (DE3). The recombinant protein with the molecular mass of 4.4×104 was successfully expressed in E. coli by optimizing the conditions of expression. Western-blot and IFA revealed the good immunogenicity exhibited by the recombinant protein.

参考文献/References:


[1]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. 
[2]SU J, LI S, HU X, et al. Duck egg-drop syndrome caused by BYD virus, a new tembusu-related flavivirus [J]. PLoS One, 2011, 6 (3): 1-10.
[3]YAN P, ZHAO Y, ZHANG X, et al. An infectious disease of ducks caused by a newly emerged tembusu virus strain in mainland China [J]. Virology, 2011, 417(1): 1-8.
[4]陈仕龙,陈少莺,王劭,等. 一种引起蛋鸡产蛋下降的新型黄病毒的分离与初步鉴定[J]. 福建农业学报, 2011, 26 (2): 170-174.
[5]傅光华,黄瑜,施少华,等. 鸡黄病毒的分离与初步鉴定[J]. 福建畜牧兽医, 2011, 33 (3): 1-2.
[6]李玉峰,马秀丽,于可响,等. 一种从鸭新分离的黄病毒研究初报[J]. 畜牧兽医学报, 2011, 42(6): 885-891.
[7]ALCON S, TALARMIN A, DEBRUYNE M, et al. Enzyme-linked immunosorbent assay specific to dengue virus type 1 nonstructural protein NS1 reveals circulation of the antigen in the blood during the acute phase of disease in patients experiencing primary or secondary infections [J]. J Clin Microbiol, 2002, 40 (2):376-381.
[8]ALCON-LEPODER S, SIVARD P, DROUET M T, et al. Secretion of flaviviral non-structural protein NS1: from diagnosis to pathogenesis [J]. Novartis Found Symp, 2006, 277: 233-247.
[9]邓永强,秦鄂德. 虫媒黄病毒包膜E蛋白的研究进展 [J]. 军事医学科学院院刊, 2006, 30 (6): 575-579.
[10]STIASNY K, BRESSANELLI S, LEPAULT J, et al. Structure of a flavivirus envelope glycoprotein in its low-pH-induced membrane fusion conformation [J]. Embo J, 2004, 23(4): 728-738.
[11]李焱,时莹,徐志凯,等. 登革病毒E蛋白入胞机制研究现状[J]. 热带医学杂志, 2010, 10 (2): 221-222.
[12]黄欣梅,李银,赵冬敏,等. 新型鹅黄病毒JS804毒株的分离与鉴定[J]. 江苏农业学报, 2011, 27(2): 354-360.
[13]苏敬良. 鸭的新型黄病毒BYD引起的产蛋下降综合征[J]. 兽医导刊, 2011(4): 27-29.
[14]PLETNEV A G, BRAY M, HANLEY K A, et al. Tick-borne Langat/mos-quito-borne dengue flavivirus chimera, a candidate live attenuated vaccine for protection against disease caused by members of the tick-borne encephalitis virus complex: evaluation in rhesus monkeys and inmosquitoes [J]. J Virol, 2001, 75(17):8259 -8267.
[15]LI J, BHUVANAKANTHAM R, HOWE J, et al. The glycosylation site in the envelope protein of WestNile virus (Sarafend) plays an important role in replication andmaturation processes [J]. J Gen Virol, 2006, 87(3): 613-622.〖ZK）〗〖FL）〗

备注/Memo

备注/Memo:
收稿日期：2014-11-11 基金项目：国家自然科学基金项目（31172345）;江苏省自然科学基金项目（BK2012376）；江苏省农业科学院基本科研业务专项项目[ZX（15）4005] 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和致病分子机制研究。（Tel）025-84390047；（E-mail）zhaodongmin126@126.com 通讯作者：李银，（E-mail）muziyin08@163.com
更新日期/Last Update: 2015-06-30