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Nuclear receptor analysis can be performed by traditional means using a minimal promoter vector with nuclear receptor response elements upstream. Or 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, these methods require a cell line with the appropriate endogenous nuclear receptors, so 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 activation sequence upstream of a minimal promoter. The ligand binding domain is responsible for binding, dimerization and interactions with co-activator or co-repressors. The one-hybrid method allows working with any cell line and nuclear receptor you desire.
