hOP-1全长基因在CHO细胞的表达及活性检测  

Determination of hOP-1 full-length gene expression in CHO cells and biological activity of rhOP-1

作　　者：岳玲[1] 史俊南[2] 柴玉波[3] 孙叶方[2] 荫俊[4] 文玲英[2] 

机构地区：[1]第三军医大学大坪医院野战外科研究所口腔科,重庆400042 [2]第四军医大学口腔医学院牙体病科,西安710032 [3]第四军医大学生物化学与分子生物学教研室,西安710032 [4]军事医学科学院微生物流行病学研究所,北京100071

出　　处：《重庆医学》2007年第10期952-954,共3页Chongqing medicine

摘　　要：目的对hOP-1全长基因在真核CHO细胞的表达产物进行鉴定及活性检测,为临床和科研工作获得稳定高活性OP-1蛋白质提供依据。方法采用ELISA、SDS-PAGE和Western Blot免疫印迹法鉴定目的基因表达产物;并进行表达产物小鼠肌肉内埋置活性成骨实验。结果经ELISA、SDS-PAGE、Western Blot鉴定筛选,得到稳定表达hOP-1的CHO细胞株。小鼠肌肉内埋植实验,证实有骨细胞形成。结论hOP-1全长基因转染真核CHO细胞,表达产物rhOP-1具有良好的生物学活性。Objective To determine the expression product of human osteogenic protein-1（hOP-1） full-length gene in CHO cells and the biological activity of recombinant hOP-1 in order to provide basis for obtaining stable and highly active OP-1 protein for basic and clinical studies. Methods ELISA, SDS-PAGE, and Western-blot were used to identify the expression of purpose gene and the biological activity of recombinant OP-1. Recombinant OP-1 combined with ceramic carrier was implanted into mouse muscle and heterotopic bone formation was observed. Results CHO cell strain of stably expressed hOP-1 was obtained through identification and screening with ELISA, SDS-PAGE and Western blot. The experiment of implanting expression product in mouse muscle proved heterotopic bone formation. Conclusion The apparent differentiation towards chondrite at implanting sites,indicates that recombinant OP-1 has biological activity.

关 键 词：rhOP-1全长基因 CHO细胞 生物学活性 

分 类 号：Q78[生物学—分子生物学]