rhGM-CSF基因真核表达载体的构建及其在COS-7细胞和小鼠体内的表达  

Construction of rhGMCSF cDNA and its expnession in COS7 cells and mouse body in vivo

作　　者：张述[1] 苏国富[1] 鲍云华 芮贤良[1] 叶棋浓[1] 吴进冬 隋拥君 王恒梁[1] 徐永强 

机构地区：[1]北京生物工程研究所

出　　处：《军事医学科学院院刊》1996年第2期97-99,共3页Bulletin of the Academy of Military Medical Sciences

摘　　要：将ｈＧＭＣＳＦ基因克隆至真核表达质粒ｐＣＤＳ中，构建成重组质粒ｐＣＧ１。用电穿孔法将质粒ｐＣＧ１导入ＣＯＳ７细胞和直接注射小鼠骨骼肌内，ＥＬＩＳＡ检测显示质粒ｐＣＧ１转染ＣＯＳ７细胞后４８、７２、９６ｈ和肌注质粒ｐＣＧ１后ｄ１０、ｄ１５、ｄ２０、ｄ２５小鼠体内均有ｈＧＭＣＳＦ的表达。生物学活性检测显示含肌注ｐＣＧ１的小鼠血液有维持ＴＦ１细胞生长的作用。The recombinant plasmid pCG1 was constructed with the eukaryotic expression of plasmid pCDS and rhGMCSF gene. Then the pCG1 was transferred into the COS7 cells by the electroporation and injected directly into the BALB/c mouse quadriceps. The results of ELISA confirmed that hGMCSF was expressed and secreted at 48、72、96 hours after the transference of pCG1 into the COS7 cells, and at 10、15、20、25 days after injection of pCG1 into the mouse muscle in vivo. Meanwhile, the results of biological activity detection showed that the mouse serum had the function of maintaining the growth of TF1 cells. This demonstrates that the hGMCSF expressed by the transfected cells has certain biological function. This animal model can be used in a new trial for the treatment and prevention of leukopenia.

关 键 词：HGM-CSF 真核细胞 DNA 骨骼肌 质粒 

分 类 号：R394.8[医药卫生—医学遗传学] Q782[医药卫生—基础医学]