Team:ETH Zurich
From 2013.igem.org
Line 9: | Line 9: | ||
<li> | <li> | ||
<a href="/Team:ETH_Zurich/Team"><img src="/wiki/images/f/f7/Team_picture.JPG" /></a> | <a href="/Team:ETH_Zurich/Team"><img src="/wiki/images/f/f7/Team_picture.JPG" /></a> | ||
- | <p style="font-size:16px"><b>Team</b><br>We are a team of seven Bachelor- and Master Students at the ETH Zurich with backgrounds in Biotechnology, Biomedical Engineering, Neurobiology and Bioinformatics. The iGEM project is located at the Department for Biosystems and Engineering in Basel.If you're around Basel, | + | <p style="font-size:16px"><b>Team</b><br>We are a team of seven Bachelor- and Master Students at the ETH Zurich with backgrounds in Biotechnology, Biomedical Engineering, Neurobiology and Bioinformatics. The iGEM project is located at the Department for Biosystems and Engineering in Basel. If you're around Basel, make sure to visit our team's lab to play some really fun Colisweeper!</p> |
</li> | </li> | ||
<li> | <li> | ||
<a href="/Team:ETH_Zurich/Project"><img src="https://static.igem.org/mediawiki/2013/8/8f/Minetocoli3.png"/></a> | <a href="/Team:ETH_Zurich/Project"><img src="https://static.igem.org/mediawiki/2013/8/8f/Minetocoli3.png"/></a> | ||
- | <p style="font-size:15px"><b>From Minesweeper to Colisweeper</b><br>Almost all the features known from the original Minesweeper game can be found in Colisweeper as well. The gamefield is replaced by mine and non-mine colonies on | + | <p style="font-size:15px"><b>From Minesweeper to Colisweeper</b><br>Almost all the features known from the original Minesweeper game can be found in Colisweeper as well. The gamefield is replaced by mine and non-mine colonies on an agar plate. Due to AHL communication and signal processing, they express different hydrolase enzymes. Through pipetting, the colorless substrate solutions turn into different colors which indicates the role of the played colony. You always have the possibility to mark colonies as mines with flags using the second substrate solution. |
</p> | </p> | ||
</li> | </li> | ||
Line 21: | Line 21: | ||
<map id="imagemap2" name="map"><area shape="poly" alt="" title="" coords="291,95,125,97,126,120,291,120,291,131,315,108,290,82,290,82" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_2" target="" /><area shape="poly" alt="" title="" coords="577,163,547,171,524,205,525,250,543,288,573,299,607,284,624,250,625,211,611,182,591,164" href="https://2013.igem.org/Team:ETH_Zurich/Project" target="" /><area shape="poly" alt="" title="" coords="483,331,454,317,404,313,351,335,330,375,355,411,394,431,438,432,481,420,508,391,509,364,501,345,501,345" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_3" target="" /><area shape="rect" alt="" title="" coords="225,261,358,286" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_6" target="" /><area shape="circle" alt="phoa" title="PhoA" coords="116,340,19" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_7" target="" /><area shape="circle" alt="GusA" title="GusA" coords="183,324,21" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_7" target="" /><area shape="circle" alt="LacZ" title="LacZ" coords="158,365,22" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_7" target="" /><area shape="circle" alt="LacZ" title="LacZ" coords="104,398,20" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_7" target="" /><area shape="circle" alt="Aes" title="Aes" coords="189,414,22" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_7" target="" /><!-- Created by Online Image Map Editor (http://www.maschek.hu/imagemap/index) --></map> | <map id="imagemap2" name="map"><area shape="poly" alt="" title="" coords="291,95,125,97,126,120,291,120,291,131,315,108,290,82,290,82" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_2" target="" /><area shape="poly" alt="" title="" coords="577,163,547,171,524,205,525,250,543,288,573,299,607,284,624,250,625,211,611,182,591,164" href="https://2013.igem.org/Team:ETH_Zurich/Project" target="" /><area shape="poly" alt="" title="" coords="483,331,454,317,404,313,351,335,330,375,355,411,394,431,438,432,481,420,508,391,509,364,501,345,501,345" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_3" target="" /><area shape="rect" alt="" title="" coords="225,261,358,286" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_6" target="" /><area shape="circle" alt="phoa" title="PhoA" coords="116,340,19" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_7" target="" /><area shape="circle" alt="GusA" title="GusA" coords="183,324,21" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_7" target="" /><area shape="circle" alt="LacZ" title="LacZ" coords="158,365,22" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_7" target="" /><area shape="circle" alt="LacZ" title="LacZ" coords="104,398,20" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_7" target="" /><area shape="circle" alt="Aes" title="Aes" coords="189,414,22" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_7" target="" /><!-- Created by Online Image Map Editor (http://www.maschek.hu/imagemap/index) --></map> | ||
- | <p style="font-size:15px"><b>Information Processing</b><br> Our non-mine cells are able to distinguish between different concentrations of AHL and to translate this information into the expression of different sets of enzymes. They use LuxR promoters with different sensitivities as highpass filters. The promoters were created using site-saturation mutagenesis. Through | + | <p style="font-size:15px"><b>Information Processing</b><br> Our non-mine cells are able to distinguish between different concentrations of AHL and to translate this information into the expression of different sets of enzymes. They use LuxR promoters with different sensitivities as highpass filters. The promoters were created using site-saturation mutagenesis. Through mutation of the LuxR binding sites the affinities were changed successfully.</p> |
</li> | </li> | ||
<li> | <li> | ||
<a href="https://2013.igem.org/Team:ETH_Zurich/Experiments_3"><img src="https://static.igem.org/mediawiki/2013/5/54/2013-09-16_09.30_%281%29.png"/></a> | <a href="https://2013.igem.org/Team:ETH_Zurich/Experiments_3"><img src="https://static.igem.org/mediawiki/2013/5/54/2013-09-16_09.30_%281%29.png"/></a> | ||
- | <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 | + | <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(<i>phoA</i>), β-galactosidase(<i>lacZ</i>), acetylesterase(<i>aes</i>), N-acetyl-β-glucosaminidase(<i>nagZ</i>) and β-glucuronidase(<i>gusA</i>) and their respective substrates to achieve fast and colorful outputs.</p> |
</li> | </li> | ||
<li> | <li> | ||
Line 37: | Line 37: | ||
<map id="imagemap2" name="#map2"><area shape="rect" alt="" title="PluxRmutations" coords="16,14,421,308" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_5" target="" /><area shape="rect" alt="" title="Diffusion tests" coords="441,18,855,315" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_2" target="" /><area shape="rect" alt="" title="Enzyme substrate tests" coords="23,321,444,725" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_3" target="" /><area shape="rect" alt="" title="Gameplay" coords="459,317,851,724" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_6" target="" /><!-- Created by Online Image Map Editor (http://www.maschek.hu/imagemap/index) --></map> | <map id="imagemap2" name="#map2"><area shape="rect" alt="" title="PluxRmutations" coords="16,14,421,308" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_5" target="" /><area shape="rect" alt="" title="Diffusion tests" coords="441,18,855,315" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_2" target="" /><area shape="rect" alt="" title="Enzyme substrate tests" coords="23,321,444,725" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_3" target="" /><area shape="rect" alt="" title="Gameplay" coords="459,317,851,724" href="https://2013.igem.org/Team:ETH_Zurich/Experiments_6" target="" /><!-- Created by Online Image Map Editor (http://www.maschek.hu/imagemap/index) --></map> | ||
- | <p style="font-size:15px"><b>Experiments</b><br>Starting with characterization of the AHL diffusion in agar plates we soon went on to test different sender and receiver-cell set ups using GFP as a readable output. In parallel the promoter mutagenesis and screening as well as the testing and characterizing of different substrates on the hydrolases was done.</p> | + | <p style="font-size:15px"><b>Experiments</b><br>Starting with characterization of the AHL diffusion in agar plates we soon went on to test different sender- and receiver-cell set ups using GFP as a readable output. In parallel, the promoter mutagenesis and screening, as well as the testing and characterizing of different substrates on the hydrolases was done.</p> |
</li> | </li> | ||
Line 58: | Line 58: | ||
</html> | </html> | ||
<br clear="all" /> | <br clear="all" /> | ||
+ | |||
<p>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.<br>Genetically engineered <i>Escherichia coli</i> 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. <br>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. | <p>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.<br>Genetically engineered <i>Escherichia coli</i> 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. <br>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. | ||
</p> | </p> | ||
{{:Team:ETH_Zurich/templates/footer}} | {{:Team:ETH_Zurich/templates/footer}} |
Revision as of 15:50, 16 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.