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| 猪瘟病毒E2基因在毕赤酵母中的表达 | |
| 引用本文： | 王启宇,兰邹然,姜平,张月,刘娟.猪瘟病毒E2基因在毕赤酵母中的表达[J].动物医学进展,2012(9):66-69. |
| 作者姓名： | 王启宇 兰邹然 姜平 张月 刘娟 |
| 作者单位： | 1. 南京农业大学动物医学院,江苏南京210095 2. 山东省动物疫病预防与控制中心,山东济南250022 3. 山东师范大学体育学院,山东济南250014 |
| 基金项目： | 山东省博士基金项目(2010BSC15001) |
| 摘 要： | 猪瘟是严重危害养猪业的一种烈性传染病,病死率高,被世界动物卫生组织(OIE)列为必须报告的动物疫病。E2蛋白是猪瘟病毒中最主要的保护性抗原,因此,围绕E2的基因工程疫苗研究已成为热点。本研究以含有猪瘟E2基因的重组质粒pMD18-T-E2为模板,设计一对特异性引物扩增去除跨膜区的E2基因,将PCR产物插入巴斯德毕赤酵母表达载体pPICZαA中,构建重组质粒pPICZαA-E2,将该质粒用SacⅠ酶切线性化后,电穿孔导入巴斯德毕赤酵母X-33中,经Zeocin筛选得到高拷贝转化子,通过甲醇诱导表达、SDS-PAGE和Western bolt试验验证,结果表明E2蛋白在酵母中获得成功表达。 |
| 关 键 词： | 猪瘟 E2蛋白 毕赤酵母 表达 |
Expression of CSFV E2 Gene in Pichia pastoris |
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| WANG Qi-yu,LAN Zou-ran,JIANG Ping,ZHANG Yue,LIU Juan.Expression of CSFV E2 Gene in Pichia pastoris[J].Progress In Veterinary Medicine,2012(9):66-69. | |
| Authors: | WANG Qi-yu LAN Zou-ran JIANG Ping ZHANG Yue LIU Juan |
| Institution: | 1.College of Veterinary Medicine,Nanjing Agricultural University,Nanjing,Jiangsu,210095,China; 2.Shandong Provincial Animal Disease Prevention and Control Center,Jinan,Shandong,250022,China; 3.College of Physical Culture,Shandong Normal University,Jinan,Shandong,250022,China) |
| Abstract: | Classical swine fever(CSF) is a highly contagious disease with high death rate,and usually causes huge loss in pig industry,which is classified in List A disease of OIE.E2 protein is the main protective antigen of CSF,so considerable research has focused on genetically engineering vaccines based on E2.In this study,so as to provide technological support in producing CSFV vaccine,we used the recombinant plasmid pMD18-T-E2 as the PCR template and designed one pair of primers to amplify the E2 gene without TMR.The PCR products were inserted into secretory Pichia pastoris expression vector pPICZαA,then the vector was linearised and transformed into Pichia pastoris X33 by electrotransformation.Transformants with high copies were acquired when selected under Zeocin and were induced with methonl.SDS-PAGE and Western blot analysis showed that the recombinant E2 protein was successfully expressed in Pichia pastoris,which laid the foundation for developing the subunit vaccine. |
| Keywords: | Classical swine fever protein E2 Pichia pastoris expression |
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