Abstract: The present invention relates to single and dual-reporter luminescence assays utilizing general and specific reagents to quench enzyme-mediated reactions. In one embodiment of the invention, a reagent is added to the assay which non-specifically quenches enzyme-mediated luminescent reactions. In another embodiment of the invention, a reagent is added to the assay which simultaneously quenches one enzyme-mediated luminescent reaction while activating another distinct enzyme-mediated luminescent reaction. An assay kit containing specific quench reagents, and the reagents themselves are also disclosed.

Abstract: A modified form of beetle luciferase, which has been engineered for improved genetic reporting, is disclosed. The modified form contains one or more new features. Chief among these is removal of the peroxisomal translocation sequence to yield a cytoplasmic form of the enzyme. Other changes include removal of potentially interfering restriction sites and genetic regulatory sites from the gene, improvement of the codon usage for mammalian cells. The modified luciferase reporter enzyme is also devoid of potential N-glycosylation targets to minimize post-translational modification and remains in the cytoplasm of host cells to optimize substrate availability.