牛FcγRⅠ(CD64)GST融合蛋白在大肠杆菌中的表达  

Expression of Cattle GST-FcγRⅠ Fusion Protein in E.coli

作　　者：张改平[1] 乔松林[2] 李青梅[1] 王丽[1] 张红[1] 郭军庆[1] 王选年[1] 夏春[2] 

机构地区：[1]河南省动物免疫学重点实验室,河南郑州450002 [2]中国农业大学动物医学院,北京100094

出　　处：《河南农业科学》2005年第6期79-82,共4页Journal of Henan Agricultural Sciences

基　　金：国家杰出青年基金项目 (30 12 5 0 35 ) ;国家自然科学基金重点项目 (30 2 30 2 70 )

摘　　要：采用PCR技术从牛巨噬细胞cDNA文库中扩增了牛的高亲和力IgGFc受体FcγRⅠ,测序结果显示与已报道的牛FcγRⅠ序列一致。在牛FcγRⅠ成熟蛋白胞外环区两端分别合成带有BamHⅠ、XhoⅠ限制酶切位点的引物,用PCR扩增后亚克隆到GST融合蛋白表达载体pGEX - 6P- 1,转化宿主菌BL2 1(DE3) ,经IPTG诱导,SDS -PAGE分析表达了N端带有GST的融合蛋白,Western -blotting试验表明融合蛋白在变性条件下能和牛IgG结合。GST -FcγRⅠ融合蛋白的成功表达为研究牛IgG和受体的相互作用机制及其介导的免疫反应打下了基础。Cattle FcγRⅠcDNA was cloned from bovine macrophage cDNA library by PCR. Sequencing the gene showed that it was identical with the published Cattle FcγRⅠgene. To construct expression plasmid, the extracellular region of the FcγRⅠgene was ligated with GST fusion protein expression plasmid pGEX-6P-1, then transformed into BL21(DE3) E.coli strain, and induced with IPTG. SDS-PAGE indicated that the fusion protein with GST was expressed. Western blot confirmed that the denatured fusion protein could combined with bovine IgG.

关 键 词：牛FcγRⅠ(CD64) GST融合蛋白 表达 

分 类 号：Q786[生物学—分子生物学]