Establishment of Tetracycline-Inducible, Survivin-Expressing CHO Cell Lines by an Optimized Screening Method

文献类型： 外文期刊

第一作者： Zhao, Xinghui

作者： Zhao, Xinghui;Yu, Yingqun;Guo, Junwei;Fu, Ling;Yu, Ting;Hou, Lihua;Yi, Shaoqiong;Chen, Wei;Yu, Yingqun;Zhao, Zhanzhong

作者机构：

关键词： tetracycline-inducible expression;enhanced green fluorescence protein;survivin;apoptosis;Chinese hamster ovary cell

期刊名称：BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY （ 影响因子：2.043； 五年影响因子：2.017 ）

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收录情况： SCI

摘要： An optimized method based on tetracycline-inducible gene expression system T-REx was developed to screen and evaluate Tet repressor (Tet R)-expressing cell lines using enhanced green fluorescence protein (EGFP) as reporter gene. To verify the effectiveness of the method, two Tet R-expressing Chinese hamster ovary (CHO) cell lines, CHO-TR B2 (stringent) and B5 (less stringent), in which the EGFP genes were variantly controlled by tetracycline, were used to construct cell lines expressing the anti-apoptosis gene survivin upon induction with tetracycline. The resulting stable clones were analyzed for survivin expression. The analysis showed that all four B5-derived clones exhibited leaky survivin expression in the absence of tetracycline, while the B2-derived clones did not. DNA laddering and annexin V/PI staining assays further indicated that although tetracycline-inducible expression of survivin conferred resistance to NH4Cl- and staurosporine-induced apoptosis in both the B2- and the B5-derived stable cell lines, the B2-derived cell lines showed more stringent regulation in the absence of tetracycline. This represents successful utilization of the present screening method.

分类号： Q

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