Disruption of the MDR3 (TruMDR3) gene of T. rubrum TIMM20092 by gene replacement strategy. (A) Schematic representation of the binary TruMDR3-targeting vector pAg1-TruMDR3/T. The nptII cassette is composed of Aspergillus nidulans
trpC promoter (PtrpC), E. coli neomycin phosphotransferase gene (nptII), and the A. fumigatus
cgrA terminator (TcgrA). LB and RB, left and right borders, respectively; A, ApaI; B, BamHI; K, KpnI; P, PstI; S, SpeI. (B) Schematic representation of the TruMDR3 locus before and after homologous recombination. (C) Southern blotting. Lane 1, TIMM20092 (parent strain); lanes 2 to 6, ΔTruMDR3-11, -22, -33, -39, and -45, respectively. A 509-bp fragment of the TruMDR3 locus was amplified by PCR with a pair of the primers P35 and P43 (Table S4) and used as a hybridization probe. DNA standard fragment sizes are shown on the left.