鸡干扰素-α基因原核表达载体的构建及高效表达  

Construction and High Level Expression of Prokaryotic Expression Vector of Chicken Interferon-α Gene

作　　者：刘成倩[1] 王静[1,2] 李红[1] 于宗幸 易建中[1] 陈磊[1,2] 

机构地区：[1]上海市农业科学院畜牧兽医研究所,上海201106 [2]上海海洋大学水产与生命学院,上海201106

出　　处：《中国畜牧兽医》2014年第4期47-50,共4页China Animal Husbandry & Veterinary Medicine

基　　金：上海市科学技术委员会重点科技攻关项目(12391901900)

摘　　要：干扰素(IFN)-α因其具有很强的抗病毒活性而备受关注。本研究根据GenBank上公布的鸡IFN-α核酸序列设计1对引物,用PCR方法扩增出鸡IFN-α基因,并将其克隆到原核表达载体pET-32a及pET-30a-DsbA上,得到重组质粒pET-32a-IFN-α及pET-30a-DsbA-IFN-α。将重组质粒转化大肠杆菌Transetta(DE3)感受态细胞,经SDS-PAGE电泳及Western blotting分析,结果表明pET-30a-DsbA-IFN-α表达量较高,表达的融合蛋白分子质量约40.4ku,表达产物主要以包涵体形式存在,且表达的融合蛋白能与His标签单克隆抗体发生特异性反应。Interferon （IFN）-a had received much more attention for its comprehensive antiviral activity. According to the relevant sequence from GenBank, a pair of specific primers was designed for amplifying chicken IFN-a gene with PCR method. The IFN-a gene was cloned into prokaryotic expression vector pET-32a and pET-30a DsbA and the protein＇s expression was i- dentified by SDS-PAGE. Immunity characteristic of the protein was analyzed by Western blotting. The cloned pET-30a-DsbA- IFN-a/E. coli was found to produce a 40.4 ku protein at the high level. The expression products existed mainly in the form of inclusion body. These expression products could specificly react with His tag monoclonal antibody.

关 键 词：鸡 干扰素-A 原核表达 

分 类 号：Q78[生物学—分子生物学]