人促红细胞生成素受体胞外区基因的克隆及其在大肠肝菌中的表达

以人胎肝为材料，通过ＰＴ－ＰＣＲ的方法扩增出人促红细胞生成素受体（ｈＰｅｏＲ）的胞外区基因。将获得的或熟体胞外区基因起始密码子改构后克隆原核表达载体ｐＢＶ２２０中，进行原核温控诱导表达。表达产物经蛋白Ｎ端测序及Ｗｅｓｔｅｒｎ－ｂｌｏｔ实验证实表达产物是ｈＥｐｏＲ胞外区蛋白。利用上罐发酵培养获得的包涵体蛋白经复性纯化后，体外生物学活性检测表明表达产物可特 抑制ＴＦ－１细胞在Ｅｐｏ刺激下的生长，证实了

Cloning cDNA of Extracellular Domain of Human Erythropoietin Receptor and its Expression in Escherichia coli

Human erythropoietin receptor (hEpoR) plays an important role in regulating the red blood cell production by promoting the proliferation and differentiation of RBC from erythroid precursors. hEpoR is a transmembrane protein,and its extrocellular domain (sEpoR) is of great importance in Epo signal transduction pathway.We cloned the gene of sEpoR by RT PCR from the total RNA of human fetal liver and expressed it in E.coli after insertion of the gene in the expression vector pBV220. The cloned gene was confirmed by sequencing analysis and gene product was confirmed by both Western blot and its first 11 amino acid residues sequence of the N\|terminal. In vitro bioassay showed that the purified gene product can repress the growth of TF cells in the presence of Epo.