Team:ETH Zurich
From 2013.igem.org
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<p style="font-size:15px"><b>Hydrolase Reactions</b><br> Enabling a fast interaction with our system,the hydrolytic enzymes generated as a signal response are an important feature of our game. We used Alkaline Phosphatase(PhoA), Beta-galactosidase(LacZ), Acetyl esterase(Aes), Glycoside hydrolase(NagZ) and Beta-glucuronidase(GusA) and their respective substrates to achieve fast and colorful outputs.</p> | <p style="font-size:15px"><b>Hydrolase Reactions</b><br> Enabling a fast interaction with our system,the hydrolytic enzymes generated as a signal response are an important feature of our game. We used Alkaline Phosphatase(PhoA), Beta-galactosidase(LacZ), Acetyl esterase(Aes), Glycoside hydrolase(NagZ) and Beta-glucuronidase(GusA) and their respective substrates to achieve fast and colorful outputs.</p> | ||
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<p style="font-size:15px"><b>The Model</b><br>We constantly tried to validate and improve our system using a spatio-temporal model. | <p style="font-size:15px"><b>The Model</b><br>We constantly tried to validate and improve our system using a spatio-temporal model. | ||
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Revision as of 13:53, 12 September 2013
Colisweeper is an interactive, biological version of the Minesweeper computer game, based on luxI/luxR quorum sensing and chromogenic enzymatic reactions. The goal is to clear an agar “minefield” without detonating mines.
Genetically engineered Escherichia coli colonies are used as sender-cells (mines) and receiver-cells (non-mines). Mines secrete the signaling molecule N-(3-oxohexanoyl)-l-homoserine lactone (OHHL) whereas non-mines process the signal. To distinguish between OHHL-levels, a library of PLuxR promoters with various sensitivities was created through site-saturation mutagenesis. High-pass filters were constructed to control the expression of different orthogonal hydrolases in non-mines, depending on the number of surrounding mines.
Additionally, the mines express their own hydrolase.A spatiotemporal reaction-diffusion model was established to evaluate and improve the system. To play Colisweeper, a colorless substrate solution is pipetted onto a colony of choice. The result is a defined color change within minutes, allowing identification of the played colony and the number of mines surrounding it.