转人生长激素鼠胚成纤维细胞的暂态表达方法的初步确立

探讨作为转基因克隆动物核供体的、不具备分泌人生长激素（hGH）功能的转hGH鼠胚胎成纤维细胞（tEF）体外表达人生长激素的简便的暂态表达方法。首先,转染,3d后筛选出G418,与无hGH分泌的人乳腺癌细胞株（MCF-7）在聚乙二醇（PEG）作用下融合,培养1～2d,放射免疫分析方法检测相同数量的MCF-7组、转染MCF-7组、tEF组以及融合细胞共4组培养液中hGH的表达。结果显示tEF组和MCF-7组均无hGH表达;二者的融合细胞组培养液中hGH表达量可高达0.84mIU/L。可见,不表达hGH的tEF与MCF-7融合形成的杂种细胞,可作为暂态表达系统检测转基因细胞的表达。

Establishment of a new transient expression way of production of human growth hormone

To establish a new transient expression way of production of human growth hormone,the clone of embryonic fibroblast was selected and fusion with transfected with specific expression vector pLNCX-βLG-hGH by lipofectine.3 days later,positive clones with G418 were screened.Then these fused transgenic cells were mixed with MCF-7（the cell strain of human mammary cancer） with PEG and cultured for 2 days to produce the hybrid cells.Radio-immunity analysis was used to detect the hGH expression of hybrid cells.The production of human growth hormone was detected from hybrid group with 0.84mIU/L.No secretion of hGH was found from transgenic mouse embryonic fibroblast and MCF-7 secrete.It indicated that hybrid cell produced by this method can be used as a transient expression way.