[1]韩凯凯,李银,黄欣梅,等.鸭坦布苏病毒囊膜蛋白抗原表位的串联表达与抗原性分析[J].江苏农业学报,2015,(01):112-116.[doi:10.3969/j.issn.1000-4440.2015.01.017]
　HAN Kai-kai,LI Yin,HUANG Xin-mei,et al.Coexpression and antigenicity of epitopes on duck Tembusu virus envelope protein E[J].,2015,(01):112-116.[doi:10.3969/j.issn.1000-4440.2015.01.017]
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鸭坦布苏病毒囊膜蛋白抗原表位的串联表达与抗原性分析()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2015年01期
页码:
112-116
栏目:
畜牧兽医·水产养殖
出版日期:
2015-02-28

文章信息/Info

Title:
Coexpression and antigenicity of epitopes on duck Tembusu virus envelope protein E
作者:
韩凯凯; 李银; 黄欣梅; 赵冬敏; 刘宇卓; 谢星星; 安凤娇
（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):
HAN Kai-kai; LI Yin; HUANG Xin-mei; ZHAO Dong-min; LIU Yu-zhuo; XIE Xing-xing; AN Feng-jiao
（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; 合成多肽; 表达; 抗原性
Keywords:
duck Tembusu virus; synthesized peptide; expression; antigenicity
分类号:
S858.32
DOI:
10.3969/j.issn.1000-4440.2015.01.017
文献标志码:
A
摘要:
在对鸭坦布苏病毒囊膜蛋白抗原表位分析的基础上，将已经筛选出的8条抗原性良好的多肽采用柔性肽拼接，转入T载体中。将酶切回收的串联基因（命名为TEM）克隆于表达载体pET32a中，获得重组表达质粒pET32a-TEM，将该质粒转化于感受态细胞BL21(DE3)中，经IPTG诱导后进行超声裂解，对融合蛋白进行可溶性分析、Western blotting分析和免疫原性分析。重组质粒pET32a-TEM的酶切和测序结果表明，TEM串联基因已成功插入到pET32a原核表达载体中；表达的pET32a-TEM融合蛋白分子质量约为38 000，以包涵体形式存在。Western blotting和小鼠免疫试验结果显示，pET32a-TEM融合蛋白与鸭坦布苏病毒阳性血清能发生特异性反应，具有良好的抗原性。串联多肽的成功表达，将为鸭坦布苏病血清学检测方法建立、新型工程疫苗的研发奠定基础。
Abstract:
Based on the analysis of the amino acid sequences of envelope protein E of duck Tembusu virus (DTMUV) strain (JS804) by bioinformatics software, eight peptides with good antigenicity were linked cloned into T vector, then the gene (named TEM) was accurately cloned into plasmid pET32a. The correct recombinant plasmid pET32a-TEM was transferred into competent Escherichia coli BL21 cells and the recombinant protein was induced and expressed with isopropyl thiogalactoside (IPTG). SDS-PAGE showed that the fusion protein was 38 000 in size and existed in the form of insoluble inclusion body. Western blotting analysis and mouse immunological results revealed the good specificity of the fusion protein. The successful expression of mass-peptide will lay the foundations for the development of serological diagnosis and effective bioengineering vaccines for duck Tembusu virus detection.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期：2014-07-14 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（BK20130710）；江苏省农业科技自主创新基金项目 [CX(12)5048] 作者简介：韩凯凯(1983-),男，河南新乡人，博士，助理研究员，主要从事家禽病毒分子生物学研究。（E-mail）hankk0917@126.com 通讯作者：李银,(E-mail)muziyin08@163.com
更新日期/Last Update: 2015-02-28