Mutagenesis Open Access Journals

Mutagenesis in model structures pursuing exposure to chemicals is engaged as an signal of genotoxicity. Mutagenesis assays also are wont to study mechanisms of DNA homeostasis. This branch focuses on disclosure of mutagenesis in prokaryotes, which boils right down to two accesses: reporter inactivation (forward mutation assay) and rotation of an inactivating mutation (reversion mutation assay). Both methods are labor intensive, involving visual screening, quantification of colonies on solid media, or resolving a Poisson distribution in liquid culture. Here, we present two reversion reporters for in vivo mutagenesis that present a quantitative output, and thus have the potential to greatly lower the amount of test chemical and labor suggested in these assays. This output is recovered by coupling a TEM β lactamase-based reversion assay with GFP fluorescence, either by placing the two genes on the same plasmid or by fusing them translationally and disturbing the N-terminus of the chimeric ORF with a stop codon. We also describe a writer aimed toward facilitating the monitoring of repeated mutagenesis in mutator strains. This writer couples two inversion markers, releasing the temporal dissolution of mutation events in time, thus providing information about the dynamics of mutagenesis in mutator strains.        

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