Improvements to Luciferase Reporter Assays for Nuclear Receptor Function Scientific Poster
Aileen Paguio, Pete Stecha, Frank Fan and Keith Wood
Promega Corporation, 2800 Woods Hollow Rd., Madison, WI 53711
Luciferase reporter assays are frequently used to study nuclear receptor function because of their wide dynamic range, low endogenous activity and ease of use. Recent improvements to pGL4 Luciferase Reporter vectors have further enhanced their utility for research and drug discovery. We used an assay containing the ligand binding domain (LBD) of a nuclear receptor fused to the Gal4 DNA binding domain (DBD) on one vector and multiple Gal4 upstream activator sequences (UAS) upstream of luciferase reporter on another vector to monitor ligand binding and transactivation. We showed that codon optimization of luciferase reporter genes increased expression levels. Incorporation of protein destabilizing sequences into luciferase leads to a larger assay dynamic range up to several hundred fold. The optimum number of UAS to generate the best response was also evaluated.
- Part# PS065
- Printed in USA.