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Rapid selection of tetracycline-controlled inducible cell lines using a green fluorescent-transactivator fusion protein.

Publication ,  Journal Article
Callus, BA; Mathey-Prevot, B
Published in: Biochem Biophys Res Commun
April 21, 1999

We describe a modification of the tetracycline-controlled expression system that facilitates the rapid identification of tetracycline-sensitive clones. The TetR/VP16 transactivator protein was tagged with the green fluorescent protein (GFP) at its N-terminus. This results in a functional transactivator which allows cells expressing high levels of the modified transactivator to be selected by fluorescent-activated cell sorting. After expansion, single cell clones that maintain a high level of GFP fluorescence can be tested for their ability to transactivate a luciferase gene under control of the Tet operator, leading to the rapid identification of clones with strong inducibility.

Duke Scholars

Published In

Biochem Biophys Res Commun

DOI

ISSN

0006-291X

Publication Date

April 21, 1999

Volume

257

Issue

3

Start / End Page

874 / 878

Location

United States

Related Subject Headings

  • Transfection
  • Trans-Activators
  • Time Factors
  • Tetracycline
  • Recombinant Fusion Proteins
  • Promoter Regions, Genetic
  • Mice
  • Luminescent Proteins
  • Luciferases
  • Herpes Simplex Virus Protein Vmw65
 

Citation

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Callus, B. A., & Mathey-Prevot, B. (1999). Rapid selection of tetracycline-controlled inducible cell lines using a green fluorescent-transactivator fusion protein. Biochem Biophys Res Commun, 257(3), 874–878. https://doi.org/10.1006/bbrc.1999.0558
Callus, B. A., and B. Mathey-Prevot. “Rapid selection of tetracycline-controlled inducible cell lines using a green fluorescent-transactivator fusion protein.Biochem Biophys Res Commun 257, no. 3 (April 21, 1999): 874–78. https://doi.org/10.1006/bbrc.1999.0558.
Callus BA, Mathey-Prevot B. Rapid selection of tetracycline-controlled inducible cell lines using a green fluorescent-transactivator fusion protein. Biochem Biophys Res Commun. 1999 Apr 21;257(3):874–8.
Callus, B. A., and B. Mathey-Prevot. “Rapid selection of tetracycline-controlled inducible cell lines using a green fluorescent-transactivator fusion protein.Biochem Biophys Res Commun, vol. 257, no. 3, Apr. 1999, pp. 874–78. Pubmed, doi:10.1006/bbrc.1999.0558.
Callus BA, Mathey-Prevot B. Rapid selection of tetracycline-controlled inducible cell lines using a green fluorescent-transactivator fusion protein. Biochem Biophys Res Commun. 1999 Apr 21;257(3):874–878.
Journal cover image

Published In

Biochem Biophys Res Commun

DOI

ISSN

0006-291X

Publication Date

April 21, 1999

Volume

257

Issue

3

Start / End Page

874 / 878

Location

United States

Related Subject Headings

  • Transfection
  • Trans-Activators
  • Time Factors
  • Tetracycline
  • Recombinant Fusion Proteins
  • Promoter Regions, Genetic
  • Mice
  • Luminescent Proteins
  • Luciferases
  • Herpes Simplex Virus Protein Vmw65