Fig 1. Deletion of gpa11 and gpa12 in M. circinelloides confirmed by Southern-blot.

A) gpa11; B) gpa12; and C) gpa11/gpa12 deletion. The 5′ and 3′ regions (1.1 kb for each) upstream and downstream from the start and stop codons, respectively, of gpa11 or gpa12 were designed to flank the pyrG selection marker for gene deletion (A and B). For the deletion of gpa11 in Δgpa12, the 5′ and 3′ regions (1.1 kb for each) upstream and downstream from the start and stop codons, respectively, of gpa11 were designed to flank leuA (C). For each diagram, the recombinant fragments that were used to transform protoplasts of the strain MU402 (pyrG⁻, leuA⁻) are shown. Molecular confirmation by Southern blotting was performed using specific probes for each gpa gene. DNA samples from transformed and parental (MU402) strains were digested with the indicated restriction enzymes (H, HincII; N, NcoI; P, PvuII).