利用乙肝核心抗原展示口蹄疫病毒2C蛋白表位  

The FMDV Protein 2C Antigen Epitope Displayed by Self-assembly Ability of HBc

作　　者：魏婕[1] 魏玉荣[1] 薛英[1] 米晓云[1] 苗书魁[1] 易忠[1] 马文戈[1] 郭会玲[1] 薛晶[1] 王丽 马丽 汪立群 黄炯[1] 

机构地区：[1]新疆畜牧科学院兽医研究所,乌鲁木齐830000 [2]新疆维吾尔自治区动物卫生监督所,乌鲁木齐830063

出　　处：《新疆农业科学》2014年第12期2274-2278,共5页Xinjiang Agricultural Sciences

基　　金：新疆维吾尔自治区自然基金青年基金项目(2012211B55);国家科技支撑计划(2013BAD12B04)

摘　　要：【目的】利用乙肝核心蛋白(Hepatitis B virus core protein,HBc)自我组装的能力将口蹄疫病毒(Footand-mouth disease virus,FMDV)非结构蛋白2C的抗原表位插入到HBc的主要免疫优势区(Major immunodominant region,MIR),使2C的表位展示在衣壳表面,鉴定其是否具有免疫反应性。【方法】人工合成筛选的2C蛋白表位序列,将其插入HBc的MIR区,构建原核表达质粒p△HBc2C,室温条件下,1m M IPTG过夜诱导表达蛋白,诱导物离心后SDS-PAGE凝胶电泳分别检测沉淀和上清是否有蛋白表达,Western-blot鉴定表达蛋白是否具有免疫反应性。【结果】SDS-PAGE和Western-blot鉴定,沉淀中25 k D处有特异性条带出现,与预期的23 k D蛋白相符,证明2C蛋白具有免疫反应性。【结论】利用HBc自我组装空衣壳的能力展示筛选的2C蛋白抗原表位具有免疫反应性。[ Objective] The project aims to explore the nonstructural protein 2C antigen epitope from foot - and - mouth disease virus （FMDV） by using the ability of self - assembly of Hepatitis B virus core protein （ HBc）, which was inserted into the major immunodominant region （MIR） of HBc, and it will be displayed on the capsid surface to identify immune reaction. [Method] Artificial screening of 2C protein epitope sequences, inserted in the MIR area of HBc, to construct the prokaryotic expression plasmid p A HBc2C. At room temperature, IPTG concentration should be 1 mM, protein expression induced by night, protein after tentrifugation were used to pellet and supernatant, then that is detected whether the expressed protein has immune reaction or not. [ Result ] By SDS - PAGE and Western - blot identification, there were specific bands appeared at 25 kD,which was consistent with the expected 23 kD protein. This proved that 2C proten had immune response. [ Conclusion]The ability to use HBe selfassembly of empty capsid displayed that screeningthe epitope of 2C protein had immune response.

关 键 词：FMDV HBC 抗原表位 2C蛋白 

分 类 号：S852.65[农业科学—基础兽医学]