【摘要】 用氨甲喋呤 (MTX)对稳定转染人促红细胞生成素 (EPO)基因的CHOˉ(DHFR缺陷型 )细胞株进行梯度加压 ,并使用α -MEM培养基进行培养 ,使DHFR基因在CHOˉ细胞中大量扩增 ,从而使EPO的表达量提高。经检测分析EPO的表达量 ,筛选出EPO高效表达的细胞株。结果表明 ,用MTX加压到 16× 10 - 7mol l和 6 4× 10 - 7mol l浓度时 ,收集的细胞培养液经 (NH4 ) 2 SO4 盐析、透析等处理得到EPO抽提液 ,经SDS -聚丙烯酰胺凝胶电泳 (SDS -PAGE)检测出与标准品相对应的一条带 ,WesternBlot证明该带确为EPO。更多还原

【Abstract】 The hEPO gene-containing CHO cells cultured in α-MEM medium were administrated with Methotrexate （MTX） at the gradient of concentrations to amplify the hEPO gene copies in the cells.The result showed that the high concentration-tolerating CHO cell strain for MTX treatment was screened.The collected medium from MTX-treated CHO cells was processed by salting-out of （NH₄）₂SO₄ and dialysis.The same specific band as standard EPO on the gel of SDS-PAGE was observed,which was identified to be EPO by western-blot method.更多还原