Publication Date

Summer 2018

Document Type

Paper

Department

Life and Environmental Sciences

First Instructor

Stefanie Otto-Hitt

Experiment Type

Reverse transcriptase (RT) PCR

Feature

Glycolysis

Gene

PFK1: Alpha subunit of heterooctameric phosphofructokinase

Abstract

The purpose of this experiment was to test whether alcohol induces starvation conditions in Tetrahymena thermophila. For this experiment, it was hypothesized that alcohol induces starvation due to a lack of energy available in the organism for feeding. This hypothesis was tested by monitoring food vacuole formation and expression of the PFK1 gene in Tetrahymena that were exposed to alcohol. The PFK-1 gene was chosen because its encoded protein plays an essential role in cellular metabolism. For the experiment, control and experimental cultures of Tetrahymena themophila were maintained in a nutrient rich media, with the media of the experimental group being supplemented with 1.75% ethanol. The production of food vacuoles was monitored using India ink over a course of 24 hours following the addition of alcohol. After 24 hours, RNA was extracted from the Tetrahymena and semiquantitative Reverse Transcription -Polymerase Chain Reactions (RT-PCRs) were performed to determine the expression of the PFK-1 gene. We predicted that alcohol would cause Tetrahymena themophila to starve by reducing their ability to produce food vacuoles. Furthermore, it was predicted that the inability of Tetrahymena to produce food vacuoles in the presence of alcohol would lead to a reduction in the expression of PFK-1.

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