1887

Abstract

Antisense repression was used as a method to alter gene function in the human-pathogenic fungus . The calcineurin A gene () and the laccase gene () were targeted since disruption of these loci results in phenotypes that are easy to screen (temperature sensitivity and lack of melanin, respectively). Serotype D yeasts were transformed with a plasmid containing the cDNA in an antisense orientation under the control of the inducible promoter, and serotype A yeasts were transformed with a plasmid containing the cDNA in an antisense orientation under the control of the constitutive actin promoter. The calcineurin transformants demonstrated a temperature-sensitive phenotype only when grown on galactose, and the laccase transformants had decreased melanin production. Northern blot analysis of the calcineurin antisense transformants confirmed that the inducible phenotype was associated with a decrease in the native transcript levels. Furthermore, it was possible to modestly impair growth of at 37 °C by using a 30 bp antisense oligonucleotide targeting . Antisense repression is now available as a tool for molecular studies in this organism, and may be applicable to other human-pathogenic fungi that have less amenable genetic systems.

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2002-01-01
2019-10-17
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