Figure 4: Mutagenic spectra of the MPs.
From: Development of potent in vivo mutagenesis plasmids with broad mutational spectra
(a–d) The rpoB locus of single rifampin-resistant colonies was amplified by PCR and sequenced in both clusters I (aa 451–754) and II (aa 84–401). (e) Identities of the rifampin-resistant rpoB alleles from MP1, MP4 or MP6 mutagenesis, or using XL1-Red. The MPs yielded a wide distribution of mutation types, with the diversity of alleles strongly correlating with MP potency. (f–i) SP063 phage containing a constitutive lacZ expression cassette was propagated on the indicated mutator strain under induced conditions, and mutations were analysed by high-throughput sequencing. In all cases, the number of sequenced mutations (n) is indicated for the MP and XL1-Red assays. *For d and e, all sequenced XL1-Red rifampin-resistant colonies carried the identical F505S/S531F rpoB genotype. aa, amino acid.
