Sequential disruption of CaMNT1 using the ura-blaster technique. (A) Schematic representation of the construction of the CaMNT1, ΔCamnt1::hisG-URA3-hisG and the ΔCamnt1::hisG alleles. The probe used for Southern analysis was a 500-bp EcoRI–HindIII fragment of CaMNT1. E, EcoRI. (B) Southern analysis of EcoRI-digested chromosomal DNA obtained from Camnt1 mutant strains at various stages of construction of a homozygous Camnt1 disruptant strain. Lanes: 1, parental strain C. albicans CAI4; 2, primary transformant NGY21; 3, post-FOA progeny of NGY21, NGY22; 4, secondary transformant NGY23; 5, post-FOA progeny of NGY23, NGY24.