克服位置效应提高外源基因在CHO细胞中稳定高效表达的策略  

Strategies to Improve the Stability of Protein Production from High Level Expression Recombinant CHO Cells

作　　者：周宏[1] 刘志刚[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》2006年第6期945-949,共5页Letters in Biotechnology

摘　　要：能够生产有功用的治疗性蛋白的一个重要前提是获得稳定的重组蛋白高表达细胞株,然而筛选一个能够持续稳定表达外源蛋白的重组细胞株是费时费力的过程。有多篇文献报道了重组蛋白细胞株表达的不稳定性。位置效应是高表达细胞株不稳定性的重要因素,克服或利用位置效应是当前获得稳定高表达重组蛋白细胞株的有效途径。为解决外源基因插入的随机性所带来的不可预知的后果,可以事先在CHO细胞基因组中筛选转录热点区域,再通过位点特异性或同源重组的方式,实现外源基因的定点整合。各种调节位置效应的DNA元件陆续被发现,可以利用它们去调控基因表达及增加细胞株的稳定性。One of the most important criteria for successful generation of a therapeutic protein from a recombinant cell is to obtain a cell line that maintains stability of production. However, selection of a cell line that sustains stability of production over the required time period may be difficult to achieve during development of a therapeutic protein. There are several studies in the literature that have reported on the instability of protein production from recombinant cell lines. Position effect is the key effecter of instability of high efficiency recombinant cell lines. To conquer or exploit position effect is an effective way to obtain a high level expression recombinant cell line. One of the most promising methods involves targeted integration of gene（s） into areas （hot spot for gene transcription） that are thought to be relevant for high expression by site-specific integration or homologous recombination. Various DNA elements have been discovered that can modulate the position effect and hence may be important in controlling expression levels and stability of protein production.

关 键 词：CHO细胞 稳定性 重组蛋白 高效表达 

分 类 号：Q786[生物学—分子生物学]