| 伪狂犬病病毒gB蛋白抗原表位基因串联构建及原核表达 |
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| 引用本文： | 王雨,吉艺宽,程艺,向柯宇,张宝石,琚春梅.伪狂犬病病毒gB蛋白抗原表位基因串联构建及原核表达[J].中国畜牧兽医,2015,42(4):810-815. |
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| 作者姓名： | 王雨 吉艺宽 程艺 向柯宇 张宝石 琚春梅 |
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| 作者单位： | 华南农业大学兽医学院, 广州 510642 |
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| 基金项目： | 国家自然科学基金(31001074) |
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| 摘 要： | 本试验通过自行设计两段引物,利用重叠延伸PCR方法,将伪狂犬病病毒(PRV)gB基因两段优势抗原表位序列串联,克隆至pMD18-T载体,测序正确后双酶切连接至pET-32a(+)载体,构建重组质粒;重组质粒转化入大肠杆菌BL21(DE3)受体菌,经IPTG诱导后,通过SDS-PAGE和Western blotting检测融合蛋白表达情况。经检测,诱导后的重组蛋白获得表达,重组蛋白大小约为54ku,其中串联蛋白大小约为34ku。该重组蛋白可与伪狂犬病病毒gB蛋白单克隆抗体发生特异性反应,表明重组蛋白的抗原性良好。
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| 关 键 词： | 伪狂犬病病毒 gB基因 抗原表位 串联基因表达 |
| 收稿时间： | 2014-10-08 |
Construction of Tandem Epitopes of gB Gene of Pseudorabies Virus and its Prokaryotic Expression |
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| WANG Yu,JI Yi-kuan,CHENG Yi,XIANG Ke-yu,ZHANG Bao-shi,JU Chun-mei.Construction of Tandem Epitopes of gB Gene of Pseudorabies Virus and its Prokaryotic Expression[J].China Animal Husbandry & Veterinary Medicine,2015,42(4):810-815. |
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| Authors: | WANG Yu JI Yi-kuan CHENG Yi XIANG Ke-yu ZHANG Bao-shi JU Chun-mei |
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| Institution: | College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China |
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| Abstract: | The two dominant epitope sequences of pseudorabies virus (PRV) gB gene were joined together by overlap-extension PCR method with two pairs of primers. The products were inserted into pMD18-T vector. After properly sequenced, the multi-epitope sequence was linked with pET-32a(+) vector, and then transformed into BL21(DE3). By being induced with IPTG, the targeted protein was detected by SDS-PAGE and Western blotting. The results showed the fusion protein was successfully expressed with the size of 54 ku including 34 ku tandem protein and could be combined with monoclonal antibody against gB which showed that the expressed protein had good antigenicity. |
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| Keywords: | pseudorabies virus (PRV) gB gene epitope tandem gene expression |
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