Fig. 4

PCR verification of Bp 576a ∆purM::FRT2-ble-FRT2, ble selection marker removal, and adenine auxotrophy screening. a Agarose gel showing the PCR product from the wild-type (WT) 576a purM region and the shift-up after deletion by insertion of the FRT2-ble-FRT2 cassette in three mutants (mut1–3). b Agarose gel showing the PCR product from the wild-type (WT) 576a purM region, the shift to a larger size after deletion of an internal purM fragment and insertion of the FRT2-ble-FRT2 cassette and the shift to a smaller size after Flp-excision of the ble cassette in two isolates. Genotypes and phenotypes are indicated below panels (a) and (b). Patch plates showing the phenotype of 576a wild-type (WT) and 576mn patched on minimal glucose (MG) + thiamine media (c) and on MG + adenine + thiamine media (d)