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| 水泡性口炎病毒N蛋白原核表达及免疫原性分析 | |
| 引用本文： | 张雪,娄丽,陈阳,周晓丽,刘钊,贾赟,孙颖杰.水泡性口炎病毒N蛋白原核表达及免疫原性分析[J].中国畜牧兽医,2017,44(9):2593-2597. |
| 作者姓名： | 张雪 娄丽 陈阳 周晓丽 刘钊 贾赟 孙颖杰 |
| 作者单位： | 1. 辽宁出入境检验检疫局, 大连 116001; 2. 丹东出入境检验检疫局, 丹东 118000; 3. 四川出入境检验检疫局, 成都 610000 |
| 基金项目： | 十二五国家科技支撑计划"外来与新发动物疫病筛查与鉴定技术研究与示范"（2013BAD12B01） |
| 摘 要： | 本研究旨在克隆和表达水泡性口炎病毒（vesicular stomatitis virus，VSV）特异性抗原N蛋白，进而纯化并分析其免疫原性。根据GenBank中已发表的VSV基因组N基因序列，分别合成VSV两种不同血清型的N基因，经序列对比分析后，设计合成1对特异性引物，PCR扩增获得约1 300 bp的N基因片段，将目的片段亚克隆至pCold Ⅰ原核表达载体中，经IPTG诱导表达后，采用Ni-NTA树脂亲和层析法纯化重组N蛋白。SDS-PAGE分析表明，N基因在大肠杆菌中得到表达，蛋白大小约为50 ku；Western blotting检测结果表明，该重组蛋白与VSV多克隆抗体发生特异性反应。本试验成功构建了VSV-IND和VSV-NJ的原核表达载体，实现了N蛋白在大肠杆菌中的可溶性表达，纯化后的重组蛋白具有良好的免疫原性。 |
| 关 键 词： | 水泡性口炎病毒(VSV) N蛋白 原核表达 |
| 收稿时间： | 2017-02-27 |
Prokaryotic Expression and Immunogenicity Analysis of N Protein from Vesicular Stomatitis Virus |
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| ZHANG Xue,LOU Li,CHEN Yang,ZHOU Xiao-li,LIU Zhao,JIA Yun,SUN Ying-jie.Prokaryotic Expression and Immunogenicity Analysis of N Protein from Vesicular Stomatitis Virus[J].China Animal Husbandry & Veterinary Medicine,2017,44(9):2593-2597. | |
| Authors: | ZHANG Xue LOU Li CHEN Yang ZHOU Xiao-li LIU Zhao JIA Yun SUN Ying-jie |
| Institution: | 1. Liaoning Entry-exit Inspection and Quarantine Bureau, Dalian 116001, China; 2. Dandong Entry-exit Inspection and Quarantine Bureau, Dandong 118000, China; 3. Sichuan Entry-exit Inspection and Quarantine Bureau, Chengdu 610000, China |
| Abstract: | This study was aimed to clone and express the specific antigen nucleoprotein (N) of vesicular stomatitis virus (VSV), and then purify and analyze its immunogenicity. Based on published VSV genome N gene sequence in GenBank, two kinds of N genes of VSV with different serotypes were synthesized, respectively. After sequence analysis, one pair of specific primers was designed and synthesized, N gene fragment with about 1 300 bp length was amplified by PCR, and subcloned into pCold Ⅰ expression vector. The recombinant N protein was induced with IPTG and purified by Ni-NTA. The results of SDS-PAGE showed that the N gene was successfully expressed in E. coli and the molecular weight of protein was 50 ku; The results of Western blotting showed that this recombined protein specifically reacted with polyclonal antibody serum of VSV. The recombinant vector with VSV-IND and VSV-NJ were successfully constructed, and the N protein was solubly expressed in E. coli, and the purified protein demonstrated promising immunogenicity. |
| Keywords: | vesicular stomatitis virus (VSV) N protein prokaryotic expression |
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