Figure 1

Constructs for transformation of B. cinerea. (a) bR knockout construct is based on the work of Shafran and colleagues [13] and contains two flanking regions of the bR gene (bR 3' and bR 5') and in between the Hyg^r cassette as selection marker. Homologous recombination with genomic DNA is presented (dashed lines are genomic flanking regions next to bR gene). (b) The bR gene was cloned into the pBC-Phleo plasmid between the EcoRI (upstream) and BamHI (downstream) restriction sites upstream to the Phleo^r cassette. (c) The HP1 knockout construct is composed of two flanking regions of the gene and in between a Hyg^r cassette as selection marker. The relative location of primers which were used to verify transformation is marked by arrows and numbers (detailed in Methods, primer sequences are listed in Table 1).