Bioscience, Biotechnology, and Biochemistry
Online ISSN : 1347-6947
Print ISSN : 0916-8451
Biochemistry & Molecular Biology Regular Papers
Establishment of Tetracycline-Inducible, Survivin-Expressing CHO Cell Lines by an Optimized Screening Method
Xinghui ZHAOYingqun YUZhanzhong ZHAOJunwei GUOLing FUTing YULihua HOUShaoqiong YIWei CHEN
Author information

2012 Volume 76 Issue 10 Pages 1909-1912


An optimized method based on tetracycline-inducible gene expression system T-REx was developed to screen and evaluate Tet repressor (TetR)-expressing cell lines using enhanced green fluorescence protein (EGFP) as reporter gene. To verify the effectiveness of the method, two TetR-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.

Information related to the author

This article cannot obtain the latest cited-by information.

© 2012 by Japan Society for Bioscience, Biotechnology, and Agrochemistry
Previous article Next article