Journal article
An episomal vector for stable tetracycline-regulated gene expression
Nucleic acids research, v 25(15), pp 3131-3134
01 Aug 1997
PMID: 9224615
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
The recently introduced tetracycline (Tc)-regulatable eukaryotic gene expression system based on the Escherichia coli Tn 10 tetracycline operon has proven to be a powerful tool for controlled expression of a variety of genes in vitro as well as in vivo . Control elements of this expression system are contained in two separate plasmid vectors. The tTA vector encodes a transactivator protein and the tetP vector contains a responsive operator-promoter element (tetP) that controls gene expression depending on tTA binding. Establishment of cell lines expressing a gene of interest under tetP control requires two subsequent rounds of transfection and clonal selection after each transfection. Here we describe a modification of this system in which the tetP element is placed in an episomal EBNA-based plasmid that can be stably maintained in primate but not in rodent cells. Using HeLa and human melanoma cells, we show that upon transient or stable transfection a reporter gene is expressed in a Tc-regulated manner similar to the original system. Thus, this expression system combines the advantages of episomal vectors, such as high efficiency of transfection and time-efficient selection of mass cultures, with tight control of gene expression provided by the Tc-regulatable system.
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Details
- Title
- An episomal vector for stable tetracycline-regulated gene expression
- Creators
- M Jost - The Wistar Institute of Anatomy and Biology, 3601 Spruce Street, Philadelphia, PA 19104, USAC KariU Rodeck
- Publication Details
- Nucleic acids research, v 25(15), pp 3131-3134
- Publisher
- Oxford University Press; England
- Grant note
- CA25874-16 / NCI NIH HHS
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Intensive Medical Sciences (IMS)
- Web of Science ID
- WOS:A1997XP99500025
- Scopus ID
- 2-s2.0-0030839991
- Other Identifier
- 991014877996604721
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- Web of Science research areas
- Biochemistry & Molecular Biology