A one-step approach to obtain cell clones expressing tetracycline- responsive transactivators
Despite the wide application of the tetracycline-regulated gene expression system, several drawbacks in establishing the system in in vitro-cultured cells have been described. Most of the problems are related to the obtainment of a reliable tetracycline-regulated cell clone, which often results in arduous labor. We describe here a new approach to facilitate the screening and selection of such cell clones. We have constructed a tetracycline-responsive plasmid that harbors an antibiotic resistance gene fused to the enhanced green fluorescent protein (EGFP) gene and the luciferase gene, both under the control of a bidirectional promoter. We demonstrate that the selection of tetracycline-regulated clones is highly simplified by using this plasmid. Only clones expressing the system in a functional manner are able to survive under antibiotic selection. In addition, a quick characterization of the responsiveness of the clones is possible by monitoring GFP expression in vivo. © 2004 Elsevier Inc. All rights reserved.
| Main Authors: | , , , |
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| Format: | journal article biblioteca |
| Language: | English |
| Published: |
Elsevier
2004
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| Subjects: | Tetracycline, Inducible expression, tTA transactivator, rtTA transactivator, |
| Online Access: | http://hdl.handle.net/20.500.12792/3055 http://hdl.handle.net/10261/294224 |
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