Team:TU-Eindhoven/Oud
From 2012.igem.org
The Project
This year's project will be all about Light Emitting Cells (LECs) that emit light in response to an electric stimulus. A LEC has to emit light conditionally, that is, it has to react a control signal. In our case the control signal will be an electric field that is strong enough to depolarize the plasma membrane of Saccharomyces cerevisiae cells, but weak enough for the cells to survive.
Light Emitting Cells
The plasma membrane of S. cerevisiae contains the Cch1-Mid1 channel protein complex that is homologous to mammalian Voltage Gated Calcium Channels (VGCCs). It is hypothesized that upon depolarization of the plasma membrane, calcium ions selectively enter the cytoplasm through Cch1-Mid1. Light will be emitted through the fluorescence of GECO protein, a calcium sensor that is expressed from a genetically engineered plasmid. When the calcium concentration is high the GECO proteins will be fluorescent, when the calcium concentration is low the GECO proteins will not be fluorescent. After a while the calcium concentration will drop to homeostatic levels through active transport to the yeast's vacuole and fluorescence will cease.Device for membrane depolarization
A device will be designed, built and used for applying an electric field to a suspension of live yeast cells in culture medium. Electrodes are laid out as a rectangular grid to form 'pixels' analogous to the matrix of control lines in an LCD-screen. The device is controlled by a computer with the aid of LabView. The materials used in the construction of the device are chosen such that they will not be toxic to the yeast cells. The device will not only contain an electrode matrix, but also a backlight as a source of illumination to invoke fluorescence. Filters will be used to ensure only fluorescent light originating from the yeast cells will be transmitted.Achievements in iGEM competition
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Please see iGEM official results page to see how all the teams did.
In the news
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