Figure 1: MP design and effect on mutation rate in bacteria.
From: Development of potent in vivo mutagenesis plasmids with broad mutational spectra
The mutator genes tested are colour-coded to indicate the canonical pathway being disrupted through the overexpression (a) or deletion (b) of that gene. All MPs express mutator genes from the E. coli arabinose-inducible PBAD promoter, with each gene preceded by a ribosome-binding site to enable translation from a single transcript. For a, the mutagenesis rate μbp (substitutions per base pair of the E. coli genome per generation) was calculated using rifampin resistance under uninduced (25 mM glucose, white bars) and induced (25 mM glucose+25 mM arabinose, black bars). For b, the rifampin resistance of XL1-Blue (white bar) and XL1-Red (black bar) was used to calculate μbp. In all cases, all known 77 rifampin-resistant rpoB alleles were used to calculate the mutation rate. Error bars denote the standard deviation of at least three biological replicates.
