Comprehensive Candida albicans and Saccharomyces cerevisiae Deletion Libraries

The C. albicans deletion library will be made available through the Fungal Genetics Stock Center (http://www.fgsc.net/). All C. albicans deletion strains were constructed in strain SN152 using auxotrophic marker cassettes targeted with long-flanking homology, as previously described [23] (link). All deletions were verified by diagnostic PCR of the flanks surrounding the introduced markers. The absence of the gene targeted for deletion was further verified by attempting to amplify a small internal fragment of the ORF. For a successful deletion, this intra-ORF PCR yielded no product while a wild-type control yielded a strong product. The strain background of the deletion strains was arg4Δ/arg4Δ, leu2Δ/leu2Δ, his1Δ/his1Δ, URA3/ura3Δ, IRO1/iro1Δ, with HIS1 and LEU2 function restored by the auxotrophic marker introduced at the targeted transcriptional regulator. A ‘wild-type’ control strain was created by reintroduction of a single allele of HIS1 and LEU2 (amplified from the C. albicans strain SC5314) into the parent strain. Composition of the media used for phenotyping is described in Text S1.
All S. cerevisiae deletion strains were obtained from the Saccharomyces Genome Deletion Project collection [11] (link). All S. cerevisiae deletion strains were from the homozygous deletion collection (MATa/α his3Δ1/his3Δ1, leu2Δ0/leu2Δ0, lys2Δ0/LYS2, MET15/met15Δ0, ura3Δ0/ura3Δ0), with the exception of the Δsko1 strain, which was haploid (MATa his3Δ1, leu2Δ0, met15Δ0, ura3Δ0). In all cases where the S. cerevisiae strain exhibited a phenotype that appeared divergent from the C. albicans ortholog(s), the S. cerevisiae strain deletion was validated using the primers suggested by the Saccharomyces Genome Deletion Project protocols.