[1]罗小暖,王 芳,崔玉东*,等.蓝舌病病毒VP2蛋白展示口蹄疫病毒中和表位的研究[J].中国预防兽医学报,2014,(09):731-734.[doi:10.3969/j.issn.1008-0589.2014.09.17]
 LUO Xiao-nuan,WANG Fang,CUI Yu-dong*,et al.Display expression of food-and-mouth disease virus neutralizing epitope on bluetongue virus VP2 protein[J].Chinese journal of preventive veterinary medicine,2014,(09):731-734.[doi:10.3969/j.issn.1008-0589.2014.09.17]
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蓝舌病病毒VP2蛋白展示口蹄疫病毒中和表位的研究
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2014年09期
页码:
731-734
栏目:
免疫学
出版日期:
2014-09-15

文章信息/Info

Title:
Display expression of food-and-mouth disease virus neutralizing epitope on bluetongue virus VP2 protein
文章编号:
1008-0589(2014)09-0731-04
作者:
 罗小暖12王 芳2崔玉东1*于 力2*
 1. 黑龙江八一农垦大学 生命科学技术学院,黑龙江 大庆 163319;
2. 中国农业科学院哈尔滨兽医研究所 兽医生物技术国家重点实验室/牛羊传染病研究创新团队,黑龙江 哈尔滨 150001
Author(s):
 LUO Xiao-nuan12 WANG Fang2 CUI Yu-dong1* YU Li2*
 1. College of Life Science and Technology, Heilongjiang Bayi Agricultural University, Daqing 163319, China;
2. Division of Livestock Infections Disease, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute,
Chinese Academy of Agricultural Sciences, Harbin 150001, China
关键词:
 蓝舌病病毒VP2蛋白O型口蹄疫病毒8E8表位杆状病毒表达
Keywords:
 bluetongue virus  VP2 protein  8E8 epitope of food-and-mouth disease virus serotype O  baculovirus expression
分类号:
S852.65
DOI:
10.3969/j.issn.1008-0589.2014.09.17
文献标志码:
A
摘要:
 为在蓝舌病病毒(BTV) VP2蛋白中鉴定展示表达外源B细胞表位的位点,本研究通过同源建模预测VP2蛋白的结构,采用SOE-PCR方法,将O型口蹄疫病毒编码保守的中和表位(8E8)序列分别插入BTV-8 VP2基因编码的两个不同loop区序列中,并分别构建了两种嵌合VP2基因。将嵌合表位编码序列的VP2基因克隆于杆状病毒转移载体pFastBac HTb中,转化感受态细胞DH10Bac,筛选阳性重组杆粒rBacmid-VP2-8E8,转染昆虫细胞Sf9,获得重组杆状病毒rBac-VP2-8E8。经western blot和间接免疫荧光验证,嵌合8E8表位的VP2蛋白在Sf9细胞中正确表达,并且8E8表位能够正确展示在VP2蛋白表面;VP2蛋白三聚体分析表明,嵌合8E8表位的VP2蛋白仍能形成三聚体。以上结果表明,BTV VP2蛋白至少存在两个插入位点,能够容纳12个氨基酸短肽的插入。该结果为BTV病毒样颗粒展示8E8表位的研究提供实验依据。
Abstract:
 To identification of the sites on the bluetongue virus (BTV) VP2 for surface expression of foreign peptides, the 3D structure of BTV-8 VP2 and the sites for accommodating foreign peptides were predicated based on homology modeling. Then, two chimeric genes of BTV-8 VP2 were amplified by SOE-PCR, which contained an inserted sequence encoding 12 amino acid residues of the O serotype food-and-mouth disease virus neutralizing epitope (8E8) within two different loop encoding sequences of BTV-8 VP2 gene, and the chimeric genes were cloned into pFastBac HTb vectors, respectively. The recombinant plasmids were transformed into competent cells of E.coli DH10Bac to construct recombinant bacmids. Two recombinant baculovirus were generated by transfection of the recombinant bacmids into Sf9 cells, respectively. The epitope-chimeric VP2 were correctly expressed in insect cells and the antigenic activity of the 8E8 epitope was detected by western blot and the indirect immunofluorescence assay. Moreover, the two epitope-chimeric VP2 still had the ability to form the multimetric structure. These data demonstrated that the two loops in VP2 tolerated the insertion of 12-amino acid peptide and the chimeric VP2 was potential to be used as vector for development of polyvalent vaccine.

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备注/Memo

备注/Memo:
收稿日期:2014-02-07
基金项目:国家自然科学基金(31201941)
作者简介:罗小暖(1989-),女,河北邢台人,硕士研究生,主要从事动物传染病的研究.
*通信作者:E-mail:cuiyudong@yahoo.comyuli1962@gmail.com
更新日期/Last Update: 2014-10-09