Nuclear receptor analysis can be performed with traditional means using a minimal promoter vector with nuclear receptor response elements upstream. Alternatively, you can use viral elements like the mouse mammary tumor virus long terminal repeat promoter to judge androgen or glucocorticoid responses (e.g., pGL4.36). In many cases, use of these methods requires a cell line with the appropriate endogenous nuclear receptors, meaning you may need different cell lines for each nuclear receptor study. A method using the principles of the yeast two-hybrid system was adapted for nuclear receptor work. The nuclear receptor ligand binding domain is fused to the GAL4 DNA binding domain and transfected with a firefly luciferase vector containing repeats of the GAL4 upstream activation sequence upstream of a minimal promoter. The ligand binding domain is responsible for ligand binding, homo- or heterodimerization and interactions with co-activator or co-repressors. The one-hybrid method allows you work with any cell line and nuclear receptor you desire.|
Robust: GAL4-based system removes background signals from endogenous receptors.More Sensitive: Optimized 9X Gal4 gives improved responses, better signal:noise ratio.Adaptable: Combination Renilla/Neomycin marker allows normalization with Dual-Luciferase(R) Assay or selectable markers for generating stable cell lines, all with one vector.Consistent: Compare or profile all nuclear receptors with a single experimental system.Faster Results: Destabilized and optimized luc2P luciferase gene allows greater sensitivity and shorter induction times.