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Access Type

WSU Access

Date of Award

January 2018

Degree Type

Thesis

Degree Name

M.S.

Department

Biochemistry and Molecular Biology

First Advisor

Robert Akins

Abstract

Candida albicans is an opportunistic pathogen whose adaptive mechanisms warrant research and understanding. In particular, it is the goal of these series of experiments to help determine the role, mechanism, and extent by which loss of heterozygosity (LOH) events are involved in C.albicans’ adaptive capabilities when exposed to a variety of stresses, be they fungicidal/fungistatic agents, heat shock, or chemical stresses, such as hydrogen peroxide exposure. Agents were chosen based on established literature for their ability to cause LOH or aneuploidy. The rates and prevalence of LOH in isolates surviving these stresses has been studied using allele-specific primers and qPCR, termed ASqPCR, in hopes of determining whether certain patterns of LOH correlate with certain types of resistance, and whether higher rates of LOH have been observed for more resistant isolates. Our research has shown little true aneuploidy, but widespread short tract LOH patterns along chromosome 5, as well as some off-site LOH on chromosome 3 relative to control samples.

If LOH patterns or prevalence alter an isolate’s phenotype in the face of stress, does this also increase a sample’s virulence? In vivo virulence studies are beyond the budget and scope of this project, but certain in vitro traits associated with virulence can be measured instead: ability to adhere to surfaces, CDR1 activity, and ability to invade tissue. Our data does not show a direct correlation, but rather that strains that experience LOH tend to have higher prevalence of altered in vitro phenotypes. This suggests further ASqPCR analysis conducted on virulent samples could corroborate our hypothesis.

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