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Review
. 2003 May;71(5):2299-309.
doi: 10.1128/IAI.71.5.2299-2309.2003.

Molecular genetic and genomic approaches to the study of medically important fungi

P T Magee  1 , Cheryl Gale, Judith Berman, Dana Davis
Affiliations
Review

Molecular genetic and genomic approaches to the study of medically important fungi

P T Magee et al. Infect Immun. 2003 May.
No abstract available

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Figures

FIG. 1.
FIG. 1.
Gene disruption with the UAU1 cassette. The UAU1 cassette, which contains ARG4 flanked by 5′ and 3′ sequences of URA3 with 530 bp of overlap, is amplified in a PCR using primers with 60-nucleotide tails and with homology to YFG1. These tails target integration to YFG1. Transforming a ura3/ura3 arg4/arg4 strain allows the selection of a heterozygote strain, YFG1/yfg1::UAU1, on synthetic medium lacking arginine. At some low frequency, the yfg1::UAU1 allele will become homozygous, one mechanism being mitotic recombination. The homozygous yfg1::UAU1/yfg1::UAU1 derivative is then able to generate an intact URA3 gene by recombination between the repeated URA3 sequences flanking the ARG4 gene. This results in a strain that is now Arg+ Ura+. Two distinct classes can be seen: double disruptants, which lack any wild-type YFG1, and trisomic strains, which contain both the yfg1::UAU1 and yfg1::URA3 alleles and retain a wild-type allele. Essential genes give rise only to trisomics, whereas nonessential genes give rise to both double disruptants and trisomics.
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