Reporter Gene Assays

Reporter gene assays represent another simple way to measure activation of a pathway involving kinase or phosphatase regulation. In these assays, the kinase-mediated transduction pathway results in the activation of a promoter that can be engineered to drive expression of reporter proteins such as green fluorescent protein, luciferase, or ^-lactamase. These assays can be run in an HTS format using standard fluorescent plate readers; however, they generally suffer the weakness of requiring long incubation (8 to 36 hr) before the signal is obtained. The long incubation reflects the time required by the promoter to initiate protein expression. Because of this, these assays can be prone to cytotoxins and non-specific effects. It is also possible that compounds can create artifacts by interacting with the reporter proteins.

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