Figure 1

ATM-transformation with the pABr1 vector. a) pABr1 (11.4 kb) vector map: Hygromycin (R), hygromycin phosphotransferase gene (hph); Kanamicin resistance gene (kanR); ToxA promoter, promoter sequence from P. tritici-repentis; SGFP, the S65T variant of the green fluorescent protein (GFP); CAMV35S cauliflower mosaic virus promoter; mGFP5, folding-enhanced green fluorescent protein variant; NOS polyA, Nopaline synthase terminator sequence; T-border (R) and T-border (L), right (RB) and left (LB) borders of A. tumefaciens T-DNA. The blue highlighted box is the plasmid region derived from pCT74. b) Images (100x magnification) of T. borchii hyphae transformed with either the pABr1 or the pBGgHg vector, plus an untransformed control (mock), obtained by phase-contrast (Nomarski) and GFP fluorescence microscopy (GFP). The type of treatment, vector and co-cultivation times are indicated on the left; a merge of the Nomarski and GFP images is shown in the rightmost panels. c) Quantification of transformed hyphae obtained with pBGgHg or pABr-1, expressed as percentage with respect to the total number of hyphae (~4500) present in each analyzed section. Data are the mean ± s.e.m. of at least five independent experiments. d) PCR amplification with ToxA-specific primers of total DNA extracted from mock-infected mycelia (lane 1), pABr1-transformed mycelia (lane 2), and pABr1/AGL-1 bacterial cells (lane 3) is shown in the upper panel. The results of PCR amplification of the same samples with kanR-specific primers are shown in the lower panel.