rhIFN-β1a cDNA在CHO细胞上的表达  

Expression of Recombinant Human IFN-β1a cDNA in CHO Cells

作　　者：李武平[1] 衣作安[1] 张成海[1] 王刚[1] 郑丽舒[1] 段招军[1] 吕宏亮[1] 侯云德[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所病毒基因工程国家重点实验室,北京100052

出　　处：《中国生物制品学杂志》2004年第5期290-293,共4页Chinese Journal of Biologicals

摘　　要：目的　在CHO细胞上表达rhIFN βcDNA。 方法　利用脂质体技术 ,将含hIFN β基因的pRC CMV IFNβ DNA载体和pSV2dhfr DNA质粒按 3∶1的比例转染CHO k1dhfr 细胞 ,后者是选择性标记。筛选在缺乏胸腺嘧啶的选择培养基生长的单个细胞克隆 ,通过MTX加压扩增与dhfr基因共转染IFNβ基因。筛选的细胞株通过大规模培养 ,收集培养液经一系列的步骤纯化。结果　能够耐受 0 .6 μmol LMTX的细胞可产生IFN 10 5unitperday 10 6cells ,CHO细胞中表达的hIFN β纯化后的抗病毒比活性可达 2 .2× 10 8IU mg。结论　建立了适合表达人干扰素Objective To express recombinant human IFN-β1a cDNA in CHO cells.Methods Mix recombinant plasmids pRC/CMV-IFNβ DNA and pSV2dhfr-DNA (used as a selective marker) at a ratio of (3∶1) and transfect into CHO-kldhfr cells by liposome technique. Screen the single clones grown in thymine-free selective medium for the pressure amplification of IFN-β gene. Subject the screened clones to large-scale culture, and collect the culture liquid for the purification by a series of procedures.Results The daily yield of IFN-β secreted by the cells resistant to 0.6 μmol/L MTX was 10~5 unit/10~6 cells.The specific activity of human IFN-β expressed in CHO cells reached 2.2×10~8 IU/mg after purification.Conclusion A CHO cell strain suitable for the expression of human IFN-β was established.

关 键 词：干扰素 中华仓鼠 卵巢细胞 表达 脂质体技术 hIFN-β基因 

分 类 号：Q78[生物学—分子生物学]