proUK-KGDW融合基因在CHO细胞中的高表达  

High Expression of proUK-KGDW Fusion Gene in CHO Cell

作　　者：杜韫[1] 刘志刚[1] 林建波[1] 徐桂清[1] 俞炜源[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071

出　　处：《生物技术通讯》2005年第4期384-386,共3页Letters in Biotechnology

基　　金：国家自然科学基金项目(30171100)

摘　　要：利用常规分子生物学技术,构建了新型高效的proUK-KGDW融合基因的分泌型哺乳动物细胞表达载体。将该载体线性化后转染CHO/dhfr-细胞,经G418筛选获得阳性克隆,然后挑取表达水平较高的克隆进行MTX加压扩增,以提高proUK-KGDW杂合体的表达水平,经2～3轮MTX加压扩增,获得多株表达水平超过10μg/(106细胞·24h)的稳定的高表达细胞株,为proUK-KGDW杂合体的制备及功能研究奠定了基础。The new expression vector for proUK-KGDW fusion gene was constructed by using routine methods of molecular biology. After the CHO/dhfr^- ceils were transfected with linearized vector, cells were subjected to selection with G418, then the proUK-KGDW expression level of different clone were analyzed. The clones with higher expression level were subjected to 2-3 round amplification with MTX, and many clones that could constantly overexpress proUK-KGDW fusion gene were obtained, and the expression level exceeded 10 μg/（106 ce11·24 h）.

关 键 词：proUK—KGDW融合基因 CHO/dhfr^-细胞 重组蛋白 高表达 

分 类 号：Q785[生物学—分子生物学]