【摘要】 目的　建立高效表达人肝细胞生长因子 (hHGF)的CHO细胞株。方法　将hHGF全长cDNA亚克隆到真核表达载体pTargetTM上 ,构建出pTargetTM-hHGF哺乳动物细胞表达质粒。通过该表达质粒和扩增质粒pSV2 dhfr对二氢叶酸还原酶基因缺失的CHO细胞 (CHO DG44)的共转染 ,经G41 8和MTX双筛选 ,建立稳定表达hHGF的CHO细胞株。该细胞株经MTX加压进行基因扩增 ,促使hHGF表达量显著提高。结果　获得的CHO细胞株hHGF表达量达到 1 1 2 μg/ml。结论　hHGF在CHO细胞中得到了高效表达 ,为制备hHGF开辟了新的途径更多还原

【Abstract】 Objective To establish a CHO cell line that is able to express human hematocyte growth factor (hHGF) at high levels.Methods Full-length human hepatocyte growth factor (hHGF) cDNA was subcloned into eucaryotic expression vector pTarget TM and mammalian cell exprssion plasmid pTarget TM -hHGF was constructed.The plasmid together with amlifying plasmid pSV2-dhfr was used to cotransfect dihydrofolate reductase (dhfr) gene-deficient CHO cells (CHO-DG44).MTX and G418 containing media were used to select hHGF-expressing cells.Results Stably hHGF-expression cell lines were established and the yield reached 1.12 μg/ml.Conclusions After double selection of G418 and MTX,stably hHGF-expression cell lines were established and increased expression levels that were obtained after selection with media containing increasing concentrations of MTX.更多还原