Microarray analysis of gene expression in ZMS1 and ZMS1/ZMS2 deletion mutants of Saccharomyces cerevisiae
Nathan Talley Alexa Romasco Erin Coleman
talleynd@jmu.edu romascan@jmu.edu colemaev@jmu.edu
Individual Project Individual Project
http://www.fullmoonbiosystems.com/Microarray/cDNASlide.htm
Abstract
In this microarray analysis, the study of gene expression patterns of three ZMS1 and one ZMS1/ZMS2 deletion mutations were examined in S. cerevisiae using two grids (7 & 8) of the microarray slide. Conversion of the sugar glucose-6-phosphate into simpler sugars by glucose-6-phosphate dehydrogenase (ZWF1) generates a significant amount of cellular NADPH which is involved with multiple enzymatic processes and acts as a primary cellular reducing agent of reactive oxygen species (Juhnke et al., 1996). ZMS1 and ZMS2 have been shown to be multi-copy phenotype suppressors of yeast with a ZWF1 deletion (∆zwf1) and may be regulating trans factors associated with alternative stress response genes (Slekar, 2008). RNA from wild type, ∆zms1, and ∆zms1/∆zms2 yeast was isolated using the Qbiogene RNAsafe kit, tested for purity and concentration, and electrophoresed on a 1.2% agarose gel to check for degradation. Samples were then labeled for microarray analysis using the Genisphere 3DNA Array 350 Detection kit, hybridized to array slides containing 70mer oligonucleotides from the entire yeast genome, and scanned at Davidson College. Data was then normalized and analyzed statistically with Magic Tool Software. As hypothesized, results indicate significant gene expression variance between ∆zms1 and ∆zms1/∆zms2 mutants with consistency among the three ∆zms1 samples studied; a total of twenty genes within grids 7 and 8 of the array were isolated. Future studies should include additional replications of the microarray analysis, verification of the results utilizing standard molecular techniques, and further statistical analysis.