Investigation of mRNA expression of ∆zms2 knockouts in Saccharomyces cerevisiae and comparison of reverse dyeing strategy.

By: Charlie Cuccherini, Ajeet Ryatt, and Chris McNair (CAC)

Individual Project: Protocol for t-test and Principle Component Analysis

By: Charlie Cuccherini

Individual Project: Protocol for Analysis of Variance (ANOVA) applied to microarrays

Abstract

Saccharomyces cerevisiae, better known as Baker’s yeast, is a well known model organism used to study Eukaryotic systems.  This study uses Baker’s yeast in order to study the effects of oxidative stress on the organism.  This particular experiment focused on gene expression in ∆zms2 mutants compared with the expression of wild type samples.  In order to examine and determine the changes in regulation for the genes in the two samples, Microarray technology was utilized.  RNA was isolated from the two different samples, and reverse transcribed into cDNA, which was then hybridized onto Microarray chips containing yeast oligonucleotides encompassing the entire genome.  These slides were read at Davidson college, and the resulting data was analyzed using ScanAlyze and MagicTool Microarray programs.  After normalizing our data using statistical tests and analyses, a number of different trends were detected.  Genes YNL130C and YMR307W were both found to be down-regulated in the mutant ΔZMS2 strain, both of which deal with cell wall components in the organism.  Also down-regulated was gene YNL021W, which encodes for a histone deacetylase activator.  The down-regulation of this gene could also be linked with the resulting down-regulation of the previously mentioned YNL130C as well.  In addition, our studies showed gene YOL063C, which has no known function in the yeast cell, to have similar up-regulated expression levels to those of gene YOL023, a known translation initiation factor found on the same chromosome.  Finally, genes YDR510 and YLL039C, both of which have activities involved in protein tagging, were both found to have up-regulated activities.  In order to further examine the correlations in expression of these genes, more studies will need to be conducted.

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