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Team:Missouri Miners/Project
From 2008.igem.org
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The purpose of this research is to use recombinant technology to culture yeast cells capable of determining the concentration of ethanol and using these cells to construct an ethanol sensor. Metabolic pathways exist for the metabolism of methanol and ethanol within some species of the Pichia taxa to include the yeast of our interest, Pichia pastoris. Alcohol oxidase (AO) appears to be the first and major enzyme produced in the methanol metabolic pathway of P. pastoris. However, if both ethanol and methanol are present, P. pastoris will utilize the ethanol before consuming the methanol. Consequently, the AOXI gene will not be expressed to produce the AO enzyme until the ethanol has been consumed. Fusing the AOXI gene promoter with the DNA sequence encoding a fluorescent protein will allow the expression of the AOXI gene to be detected. In supplying the yeast cells with ethanol and methanol simultaneously, the cells will produce the fluorescent protein once the ethanol is utilized. The concentration of ethanol can then be determined by measuring the time before fluorescence is detected. |  Your team picture |
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Overall project
The goal of this research is the manipulation of yeast cells; granting them the capability of measuring the concentration of ethanol present. This project utilizes the metabolic pathways of the yeast Pichia pastoris, which are capable of metabolizing ethanol and methanol. The enzyme, alcohol oxidase (AO), encoded in the AOXI gene appears to be the major enzyme involved in methanol metabolism. If both carbon sources are present, however, P. pastoris prefers to utilize ethanol first. This preference is controlled by the AOXI promoter. Fusing the AOXI promoter with a fluorescent protein gene will allow visible detection of the expression of AOXI. In supplying the yeast with ethanol and methanol simultaneously, the cells should produce the fluorescent protein after ethanol consumption; resulting in a visible color and fluorescence. The concentration of ethanol can be determined by measuring the time before fluorescence and in doing so, will make plausible the development of a breathalyzer device and additional sensor systems.
