Effects of Fatty Acids from Coconut and Olive Oil on Expression of the HMG1 Gene and Feeding Behavior in Tetrahymena thermophila

Abstract

HMG-CoA reductase is an enzyme involved in the biosynthetic pathways of sterols in many different organisms. The gene in Tetrahymena thermophila which codes for this protein is HMG1, and has many homologs across many other organisms, from fungi to humans. For this project, we wanted to look at the effect of external fatty acids from coconut and olive oil on the expression of HMG1 in T. thermophila. Previous research has shown that exposure to unsaturated oils result in decreased cholesterol levels. We hypothesized that if T. thermophila are introduced to saturated fats and unsaturated fats, they would exhibit a respectively higher and lower expression of HMG1, and that T. thermophila will selectively ingest unsaturated versus saturated fats when observed during a feeding behavioral assay. Cultures of T. thermophila will be maintained in a nutrient-poor NEFF media, and then experiments will be carried out on nutrient-rich SPP media. Our independent variables are the inclusion of 5% coconut and 5% olive oil in different SPP media, and our dependent variables are the expression of HMG1 and their feeding behavior. Experimental groups were incubated at 26 ? for 5 days to maximize exposure to the environmental changes. RNA extraction immediately followed to most accurately determine levels of HMG1 in the T. thermophila. We will determine the expression of HMG1 using Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR). Levels of HMG1 were predicted to increase in the presence of saturated fats (coconut oil) and decrease in the presence of unsaturated fats (olive oil).