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. 2008 Jul 28;52(10):3718–3724. doi: 10.1128/AAC.00446-08

FIG. 4.

FIG. 4.

Disruption of the CYP56 gene. (A) Scheme of gene disruption of the SAT1 cassette contained in plasmid pSFS2. caFLP, C. albicans-adapted FLP gene; caSAT1, nourseothricin resistance marker; MAL, promoter; ACT1, transcription termination sequence; FRT, FLP recombination target. Correct integration was confirmed by Southern hybridization using probes from the DIT2KF and DIT2XR upstream region. (B) Southern hybridization analysis showed that all clones (lanes 4 to 13) had excised the SAT1 flipper by FLP recombination and had knocked out the second wild-type allele, inactivating the CYP56 gene. The upper band corresponds to the wild-type-copy of CYP56 and is absent in lanes 4 to 13. The lower band corresponds to the deleted copy of CYP56. Lanes: 1, SC5314; 2, heterozygous CYP56/cyp56Δ mutant; 3, positive-control DIT2 PCR product; 4 to 13, homozygous cyp56Δ/cyp56Δ mutants.