Luciferase Assay to Study the Activity of a Cloned Promoter DNA Fragment

  • Nina Solberg
  • Stefan Krauss
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 977)

Abstract

Luciferase based assays have become an invaluable tool for the analysis of cloned promoter DNA fragments, both for verifying the ability of a potential promoter fragment to drive the expression of a luciferase reporter gene in various cellular contexts, and for dissecting binding elements in the promoter. Here, we describe the use of the Dual-Luciferase® Reporter Assay System created by Promega (Promega Corporation, Wisconsin, USA) to study the cloned 6.7 kilobases (kb) mouse (m) Tcf3 promoter DNA fragment in mouse embryonic derived neural stem cells (NSC). In this system, the expression of the firefly luciferase driven by the cloned mTcf3 promoter DNA fragment (including transcription initiation sites) is correlated with a co-transfected control reporter expressing Renilla luciferase from the herpes simplex virus (HSV) thymidine kinase promoter. Using an internal control reporter allows to normalize the activity of the experimental reporter to the internal control, which minimizes experimental variability.

Key words

Promoter Transfection Dual-Luciferase Firefly Renilla NSC 

Notes

Acknowledgment

The research was granted by the Norwegian Research Council.

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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Nina Solberg
    • 1
  • Stefan Krauss
    • 1
  1. 1.Unit for Cell SignalingOslo University HospitalOsloNorway

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