Yeast As a Model Organism To Assay the Functional Relevance of nsSNPS in Mitochondrial Disorders
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This thesis explores two yeast experiments. The first experiment, which was performed during the summers of 2011 and 2012, involved developing several yeast strains (including yeast strains with human wild-type orfeomes and yeast strains with human mutant orfeomes) and analyzing their growth rates with a flow cytometry growth competition assay. In this experiment, we hypothesized that 1) the yeast wild-type strains would grow better than the yeast deletion strains because the yeast deletion strain is lacking a growth gene and 2) the yeast strains with human wild-type orfeomes would grow better than the yeast deletion strains. The results of this experiment supported both hypotheses and validate our approach as a successful way to determine the relevance of certain nsSNPs in humans. The second yeast experiment, performed by scientist Sze Chern Lim and colleagues, involved identifying a homozygous mutation in two cousins with OXPHOS deficiency by using various sequencing techniques. After the homozygous mutation was identified, a mutant LYRM4 gene was able to partially complement for an ISD11 deletion in yeast. For this experiment, the scientists hypothesized that L-cysteine desulferase activity of NFS1 would be minimally present when co-expressed with a mutant ISD11 protein; the results of their in vitro experiments supported this hypothesis.