The Effects of Trolox (Vitamin E) on Gene Expression During Oxidative Stress in Saccharomyces Cerevisiae

The Effects of Trolox (Vitamin E) on Gene Expression During Oxidative Stress in Saccharomyces Cerevisiae

Cellular oxidative stress contributes to the production of reactive oxygen species (ROS) during metabolic processes, which may cause many illnesses including cancers. Hydrogen peroxide (H2O2) forms radicals that may react with various cellular components such as DNA, proteins, and lipids. This research focuses on the use of Trolox (vitamin E) as an antioxidant in the model organism Saccharomyces cerevisiae(yeast). The abilities of Trolox to decrease cellular damage caused during oxidative stress is investigated through two known oxidative stress genes and four genes of unknown function (suspected to have a role in controlling oxidative stress). Additionally, separate cultures will be exposed to two different concentrations of H2O2 (3% and 30%) to determine its effects on the six genes. RNA is extracted from the Trolox and H2O2treated yeast cultures. Quality and quantity of the RNA is measured using UV- spectrophotometry at the absorbance wavelengths of A260nm and A280nm as well as the A260/A280 ratio. Gel electrophoresis is used to analyze the quality of the RNA extracted by visualizing the large and small ribosomal subunit bands. A reverse transcription reaction is performed to convert mRNA to cDNA, and then Polymerase Chain Reaction (PCR) is performed to test for the presence and relative quantity of targeted genes. The size and relative brightness of the resulting bands are checked against the molecular size standard to determine the potential effects of Trolox and H2O2 treatment on selected gene expression.