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

Journal Article

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.

Full Text

Duke Authors

Cited Authors

  • Callus, BA; Mathey-Prevot, B

Published Date

  • April 21, 1999

Published In

Volume / Issue

  • 257 / 3

Start / End Page

  • 874 - 878

PubMed ID

  • 10208877

International Standard Serial Number (ISSN)

  • 0006-291X

Digital Object Identifier (DOI)

  • 10.1006/bbrc.1999.0558

Language

  • eng

Conference Location

  • United States