http://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/results/constructs&feed=atom&action=historyTeam:Valencia UPV/Project/results/constructs - Revision history2024-03-29T15:42:41ZRevision history for this page on the wikiMediaWiki 1.16.5http://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/results/constructs&diff=386482&oldid=prevIvllobel at 02:06, 18 October 20142014-10-18T02:06:18Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><ul style="margin-left: 1.5em;"> <li> <a href="#biosyn">RESULTS–CONSTRUCTS-BIOSYNTHESIS</a></li> <li> <a href="#phero">RESULTS-CONSTRUCTS-PHEROMONE RELEASE</a></li> <li> <a href="#switch">RESULTS - CONSTRUCTS-SWITCH</a></li> <li> <a href="#biosafe">RESULTS-CONSTRUCTS-BIOSAFETY</a></li></ul></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><ul style="margin-left: 1.5em;"> <li> <a href="#biosyn">RESULTS–CONSTRUCTS-BIOSYNTHESIS</a></li> <li> <a href="#phero">RESULTS-CONSTRUCTS-PHEROMONE RELEASE</a></li> <li> <a href="#switch">RESULTS - CONSTRUCTS-SWITCH</a></li> <li> <a href="#biosafe">RESULTS-CONSTRUCTS-BIOSAFETY</a></li></ul></div></td></tr>
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</table>Ivllobelhttp://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/results/constructs&diff=386449&oldid=prevIvllobel at 02:05, 18 October 20142014-10-18T02:05:55Z<p></p>
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</table>Ivllobelhttp://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/results/constructs&diff=386401&oldid=prevIvllobel at 02:05, 18 October 20142014-10-18T02:05:29Z<p></p>
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</table>Ivllobelhttp://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/results/constructs&diff=380800&oldid=prevAlquiru at 01:14, 18 October 20142014-10-18T01:14:58Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>We used GoldenBraid 2.0 for all our cloning reactions. GoldenBraid and BioBrick parts are not directly exchangeable; however, we adapted the coding sequences of the three biosynthetic genes to BioBrick standards using a <a class="normal-link-page" href="#">GoldenBraid-Biobricks translator</a> developed by the <a href="https://2014.igem.org/Team:NRP-UEA-Norwich/Project_Mo-Flipper" class="normal-link-page">NRP-UEA-Norwich</a> team. These BioBricks have been submitted to the Parts Registry.</p><br/><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>We used GoldenBraid 2.0 for all our cloning reactions. GoldenBraid and BioBrick parts are not directly exchangeable; however, we adapted the coding sequences of the three biosynthetic genes to BioBrick standards using a <a class="normal-link-page" href="#">GoldenBraid-Biobricks translator</a> developed by the <a href="https://2014.igem.org/Team:NRP-UEA-Norwich/Project_Mo-Flipper" class="normal-link-page">NRP-UEA-Norwich</a> team. These BioBricks have been submitted to the Parts Registry.</p<ins class="diffchange diffchange-inline">><br/</ins>><br/><br/></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Finally, after two binary GoldenBraid assembly steps, we obtained a multigenic construct with all three transcriptional units with the PCPS2 promoter. This construct was ready to be transferred to the plant to test the release of the pheromones.</p><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Finally, after two binary GoldenBraid assembly steps, we obtained a multigenic construct with all three transcriptional units with the PCPS2 promoter. This construct was ready to be transferred to the plant to test the release of the pheromones.</p><br/></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Finally, we assembled all three constructs into a single multigenic construct that was introduced into the plant to perform the Luciferase assay.</p><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Finally, we assembled all three constructs into a single multigenic construct that was introduced into the plant to perform the Luciferase assay.</p><br/></div></td></tr>
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</table>Alquiruhttp://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/results/constructs&diff=370368&oldid=prevIvllobel at 23:46, 17 October 20142014-10-17T23:46:41Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In order to have a tight control over the pheromone production in the plant, we implemented an inducible switch in the plant genetic circuit. This switch is induced with the presence of copper ions, activating the pheromone production pathway in the plant only when the insects mating season arrives. With this kind of inducible expression, the pheromones production metabolic cost for the plant will be reduced and the pheromone release will be always under control (<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/switch" class="normal-link-page">see Switch</a>).</p><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In order to have a tight control over the pheromone production in the plant, we implemented an inducible switch in the plant genetic circuit. This switch is induced with the presence of copper ions, activating the pheromone production pathway in the plant only when the insects mating season arrives. With this kind of inducible expression, the pheromones production metabolic cost for the plant will be reduced and the pheromone release will be always under control (<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/switch" class="normal-link-page">see Switch</a>).</p><br/></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In order to obtain a safe plant we developed a biosafety module which avoids spread of the plant’s genetic material and allows easy identification of the modified plant.</p><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In order to obtain a safe plant we developed a biosafety module which avoids spread of the plant’s genetic material and allows easy identification of the modified plant.</p><br/></div></td></tr>
</table>Ivllobelhttp://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/results/constructs&diff=370293&oldid=prevIvllobel at 23:46, 17 October 20142014-10-17T23:46:01Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In order to have a tight control over the pheromone production in the plant, we implemented an inducible switch in the plant genetic circuit. This switch is induced with the presence of copper ions, activating the pheromone production pathway in the plant only when the insects mating season arrives. With this kind of inducible expression, the pheromones production metabolic cost for the plant will be reduced and the pheromone release will be always under control (<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/switch" class="normal-link-page">see Switch</a>).</p><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In order to have a tight control over the pheromone production in the plant, we implemented an inducible switch in the plant genetic circuit. This switch is induced with the presence of copper ions, activating the pheromone production pathway in the plant only when the insects mating season arrives. With this kind of inducible expression, the pheromones production metabolic cost for the plant will be reduced and the pheromone release will be always under control (<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/switch" class="normal-link-page">see Switch</a>).</p><br/></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In order to obtain a safe plant we developed a biosafety module which avoids spread of the plant’s genetic material and allows easy identification of the modified plant.</p><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In order to obtain a safe plant we developed a biosafety module which avoids spread of the plant’s genetic material and allows easy identification of the modified plant.</p><br/></div></td></tr>
</table>Ivllobelhttp://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/results/constructs&diff=368742&oldid=prevIvllobel at 23:33, 17 October 20142014-10-17T23:33:21Z<p></p>
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</table>Ivllobelhttp://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/results/constructs&diff=368636&oldid=prevIvllobel at 23:32, 17 October 20142014-10-17T23:32:26Z<p></p>
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</table>Ivllobelhttp://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/results/constructs&diff=368550&oldid=prevIvllobel at 23:31, 17 October 20142014-10-17T23:31:50Z<p></p>
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</table>Ivllobelhttp://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/results/constructs&diff=368115&oldid=prevIvllobel at 23:28, 17 October 20142014-10-17T23:28:14Z<p></p>
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<td colspan='2' style="background-color: white; color:black;">← Older revision</td>
<td colspan='2' style="background-color: white; color:black;">Revision as of 23:28, 17 October 2014</td>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>We worked together with NRP-UEA-Norwich <del class="diffchange diffchange-inline"> </del>team to introduce their chromoproteins (Yellow and Blue) in our system to be used as a strategy of identity preservation. We received their complete transcriptional units and tested them in plant. However, their activity was not strong enough to be detected by naked eye.</p><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>We worked together with <ins class="diffchange diffchange-inline"><a class="normal-link-page" href="https://2014.igem.org/Team:NRP-UEA-Norwich"></ins>NRP-UEA-Norwich<ins class="diffchange diffchange-inline"></a> </ins>team to introduce their chromoproteins (Yellow and Blue) in our system to be used as a strategy of identity preservation. We received their complete transcriptional units and tested them in plant. However, their activity was not strong enough to be detected by naked eye.</p><br/></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>Since the provided chromoproteins did not show enough change in the plant’s colour to be detected by the naked eye, we tested an alternative strategy. We tested an anthocyanin production enhancing genetic construction in N.benthamiana. Anthocyanins are coloured compounds found in plants.</p><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>Since the provided chromoproteins did not show enough change in the plant’s colour to be detected by the naked eye, we tested an alternative strategy. We tested an anthocyanin production enhancing genetic construction in <ins class="diffchange diffchange-inline"><i></ins>N.benthamiana<ins class="diffchange diffchange-inline"></i></ins>. Anthocyanins are coloured compounds found in plants.</p><br/></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>We obtained this construction from GoldenBraid 2.0 <del class="diffchange diffchange-inline"> </del>collection. This construction expresses two transcription factors (Ant1 and JAF13) and successfully enhanced the synthesis of anthocyanins. Therefore, we propose an identity preservation strategy based in anthocyanin accumulation as part of our biosafety module.</p><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>We obtained this construction from <ins class="diffchange diffchange-inline"><a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/gb"></ins>GoldenBraid 2.0<ins class="diffchange diffchange-inline"></a> </ins>collection. This construction expresses two transcription factors (Ant1 and JAF13) and successfully enhanced the synthesis of anthocyanins. Therefore, we propose an identity preservation strategy based in anthocyanin accumulation as part of our biosafety module.</p><br/></div></td></tr>
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</table>Ivllobel