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The disruption of a specific gene inCandida albicans is commonly used to determine the function of the gene product. We disruptedAAF1 , a gene ofC. albicans that causesSaccharomyces cerevisiae to flocculate and adhere to endothelial cells. We then characterized multiple heterozygous and homozygous mutants. These null mutants adhered to endothelial cells to the same extent as did the parent organism. However, mutants with presumably the same genotype revealed significant heterogeneity in their growth rates in vitro. This heterogeneity was not the result of the transformation procedure per se, nor was it caused by differences in the expression or function of URA3, a marker used in the process of gene disruption. The growth rate among the different heterozygous and homozygous null mutants was positively correlated with in vivo virulence in mice. It is possible that the variable phenotypes ofC. albicans were due to mutations outside of theAAF1 coding region that were introduced during the gene disruption process. These results indicate that careful phenotypic characterization of mutants ofC. albicans generated through targeted gene disruption should be performed to exclude the introduction of unexpected mutations that may influence pathogenicity in mice.

Unanticipated Heterogeneity in Growth Rate and Virulence among Candida albicans AAF1 Null Mutants