URA3 as a selectable marker for disruption and virulence assessment of Candida albicans genes

The ability to generate isogenic sets of strains with mutations in a gene of interest but not in other genes by repeated use of the URA3 marker (Ura-blaster methodology) has advanced our understanding of the relationships between gene structure and function in Candida albicans. Common applications of Ura-blaster technology result in different genomic positions for the URA3 gene in strains complemented for the gene of interest compared with mutant strains. Studies using animal models of systemic candidiasis pointed to possible differences in URA3 gene expression, depending on its genomic location, which confounded interpretation of the role of the gene of interest in lethality. Positional effects on URA3 expression can be avoided by placement at a common locus in all strains used for comparison.