摘要
[目的]构建鸡IFN-γ的表达载体。[方法]采用PCR方法从pcDNA-ChIFN-γ重组质粒中扩增得到鸡IFN-γ基因,将目的基因定向克隆至能够在大肠杆菌、昆虫细胞和哺乳动物细胞中表达的pQE trisystem载体内,转化大肠杆菌M15菌株,以IPTG诱导表达8×H is-tag标签蛋白并对其进行纯化。[结果]通过SDS-PAGE检测方法表明,ChIFN-γ基因片段成功表达。[结论]鸡IFN-γ的表达载体成功构建。
[Objective]The aim was to study construction of expression vector of ChIFN-γ gene[Method]Chicken interferon-γ gene was cloned from the recombinant vector pcDNA-ChIFN-γ using PCR method.The fragment of ChIFN-γ was subcloned into pQE trisystem vector that could express in E.coli,insect cells and mammalian cells according to the current open reading frame.The recombinant vector was transformed into M15 bacteria and was induced by IPTG to express 8×His-tag protein which could be used to detect ChIFN-γ.[Result]The results of SDS-PAGE showed that ChIFN-γ protein was expressed successfully.[Conclusion]Construction of expression vector of ChIFN-γ gene was successful.
出处
《安徽农业科学》
CAS
北大核心
2011年第23期13935-13937,共3页
Journal of Anhui Agricultural Sciences
基金
湖南省教育厅科研项目(06C570)
2010年度湖南省高校创新平台开放基金项目(10K043)
湖南省重点建设学科(动物学)资助项目
湖南省高校产学研究合作示范基地---水生生物资源与利用项目
水生生物资源保育与利用省高校科技创新团队项目
关键词
鸡γ-干扰素
克隆
表达
Chicken interferon-gamma
Cloning
Expression
