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Nucleic Acids Research 2006 34(1):e1; doi:10.1093/nar/gnj001
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Published online 3 January 2006

© The Author 2006. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions{at}oxfordjournals.org

Methods Online

Inducible model for ß-six-mediated site-specific recombination in mammalian cells

Pilar Servert, Javier Garcia-Castro, Vicente Díaz, Daniel Lucas, Manuel A. Gonzalez, Carlos Martínez-A and Antonio Bernad*

Departamento de Inmunología y Oncología, Centro Nacional de Biotecnología, CSIC, Campus Universidad Autónoma de Madrid, Cantoblanco E-28049 Madrid, Spain

*To whom correspondence should be addressed. Tel: +34 91 585 4562; Fax: +34 91 372 0493; Email: abernad{at}cnb.uam.es

Received September 4, 2005. Revised December 1, 2005. Accepted December 1, 2005.

The prokaryotic ß recombinase catalyzes site-specific recombination between two directly oriented minimal six sites in chromatin-integrated substrates. Here, we demonstrate that an enhanced green fluorescent protein (EGFP)-fused version of ß recombinase (ß-EGFP) is fully active, retaining most specific activity. It is used to develop a recombination-dependent activatable gene expression (RAGE) system based on the androgen receptor (AR) ligand-binding domain (LBD). Two hybrid molecules, a direct fusion of the LBD-AR to the C-terminus of ß recombinase (ß-AR) and a triple fusion of ß-EGFP to the same ligand-binding domain (ß-EGFP-AR), were engineered and their subcellular behavior, stability and catalytic activity were evaluated. Both chimeric ß recombinase proteins showed in vivo inducible recombinogenic activity dependent on addition of an androgen receptor agonist, although the ß-AR fusion protein demonstrated more accurate ligand-dependent translocation from cytoplasm to nucleus.

Present addresses: Javier Garcia-Castro, Unidad de Oncología y Trasplante, Hospital Universitario del Niño Jesús, Avda. Menéndez Pelayo 65, E-28009 Madrid, Spain

Vicente Díaz and Manuel A. Gonzalez, Genetrix SL, Marconi 1, Parque Tecnológico de Madrid, Tres Cantos E-28760 Madrid, Spain


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