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Title: Use of mRNA differential display to investigate dimorphism in Candida albicans
Author: MacCallum, D. M.
Awarding Body: University of Aberdeen
Current Institution: University of Aberdeen
Date of Award: 1998
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The ability of Candida albicans to switch between a yeast and a hyphal for, known as dimorphism, has been shown to contribute to the virulence of this opportunistic fungal pathogen. mRNA differential display was used to investigate changes in gene expression which occur during dimorphism in C. albicans, identifying 50 cDNAs as being expressed differentially in yeast cells and hypha-induced cells. Further analysis of 22 cDNAs confirmed the differential expression of 45 % of cDNAs and demonstrated that 14 % appeared to be false positives. Confirmed differentially expressed genes included a fatty acid desaturase gene and a subtilisin-like serine protease gene, and a gene encoding subunit V of cytochrome c oxidase appeared to be a false positive. Further analysis of the C. albicans subtilisin-like serine protease showed that it was most similar to Saccharomyces cerevisiae yeast subtilisin-like protease III, a protease of unknown function. To determine the function of this gene a S. cerevisiae YSP3/ysp3 heterozygous strain was produced by targeted gene disruption. The heterozygous strain had reduced subtilism activity and was unable to sporulate, which may be due to an inability to degrade obsolete proteins and organelles. The C. albicans subtilisin-like serine protease full length ORF was obtained using inverse PCR and through a search of the Incyte C. albicans genome sequencing database. The deduced amino acid sequence of the ORF showed the typical secondary structures of subtilisin-like serine proteases, and was most similar to protease B, rather than yeast subtilisin-like protease III, of S. cerevisiae.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available