猪粒细胞-巨噬细胞集落刺激因子的原核表达和纯化  

Prokaryotic expression and purification of granulocyte-macrophage colony stimulating factor

作　　者：张亚宁[1] 赵红英 陈攀[1] 郑芳园[1] 李玉峰[1] 

机构地区：[1]南京农业大学动物医学院,江苏南京210095 [2]察布查尔县畜牧兽医站,新疆察布查尔835300

出　　处：《畜牧与兽医》2014年第4期14-17,共4页Animal Husbandry & Veterinary Medicine

基　　金：公益性行业(农业)科研专项项目(200903036-10)

摘　　要：粒细胞-巨噬细胞集落刺激因子(granulocyte-macrophage colony stimulating factor,GM-CSF)是一种具有多种生物学活性的细胞因子。本试验根据GenBank发表的猪的GM-CSF基因序列设计特异性引物,对GM-CSF基因片段进行扩增,扩增后的PCR产物经酶切后克隆入pET-28a载体。重组质粒转化入大肠杆菌杆菌BL21中经IPTG诱导表达,SDS-PAGE结果证明,重组蛋白获得了成功表达,并且主要以包涵体的形式存在。Western blot结果证明重组蛋白具有良好免疫反应性。以上结果为进一步应用重组GM-CSF作为血液干细胞动员剂提供了基础。Granulocyte-maerophage colony stimulating factor （GM-CSF） is a kind of cytokine with many biological activities. In this stud- y, according to the pig GM-CSF sequence published in GenBank, a pair of primers was designed. PCR fragment was amplified and cloned into pET28a （＋） to obtain recombinant plasmid pET28-GMCSF. Recombinant plasmid was transformed into BI21 and induced by IPTG. SDS-PAGE analysis showed that recombinant protein mainly existed as inclusion bodies. Western blot analysis showed that the recombinant protein has good reactogenicity. The above resuhs provide the basis for further application of recombinant GM-CSF as blood stem cell mobili- zation agent.

关 键 词：猪 粒细胞-巨噬细胞集落刺激因子(GM—CSF) 重组蛋白 原核表达 

分 类 号：S828[农业科学—畜牧学]