ASSAY FOR DRUG-INDUCED RECODING

摘要

A tissue culture assay for measuring drug-induced recoding in regulating cellular polyamine levels is described. A DNA construct containing the renilla luciferase gene separated by a short cloning site from the firefly luciferase gene, both under the control of a single upstream SV40 promoter is provided. The cloning site contains the portion of an antizyme gene known to contain the mRNA signals for polyamine stimulated frameshifting with the downstream firefly gene in the +1 position relative to the upstream renilla gene. A control construct is also produced with the genes in the same reading frame. Frameshifting efficiencies can be determined by comparing the ratio of firefly to renilla luciferase activity in parallel cell cultures.