http://2011.igem.org/wiki/index.php?title=Team:DTU-Denmark-2/results/Proofofconcept/fungi&feed=atom&action=historyTeam:DTU-Denmark-2/results/Proofofconcept/fungi - Revision history2024-03-28T15:55:50ZRevision history for this page on the wikiMediaWiki 1.16.0http://2011.igem.org/wiki/index.php?title=Team:DTU-Denmark-2/results/Proofofconcept/fungi&diff=158875&oldid=prevJaide at 20:04, 21 September 20112011-09-21T20:04:22Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a name="Proof of concept"></a><h1><b>Proof of concept</b></h1></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a name="Proof of concept"></a><h1><b>Proof of concept</b></h1></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Five different plasmids were constructed with the Plug 'n' Play assembly standard in order to verify the systems function in filamentous fungi. To ensure a successful transformation the backbone plasmid <a href="http://partsregistry.org/Part:BBa_K678046">pFun</a> has two NotI restriction sites flanking the device to be inserted. Hereby, the device can be cut out of the plasmid and the linearized DNA fragments can be transformed into the fungus. The devices for proof of concept are not <del class="diffchange diffchange-inline">design </del>to be inserted at a specific site in the fungal genome. Therefore, the device can be integrated at any site and with a random number of copies by non-homologous end<del class="diffchange diffchange-inline">-</del>joining (NHEJ) <del class="diffchange diffchange-inline">in the fungus</del>. This means that the possibility of the disruption of essential genes exists.<br><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Five different plasmids were constructed with the Plug 'n' Play assembly standard in order to verify the systems function in filamentous fungi. To ensure a successful transformation the backbone plasmid <a href="http://partsregistry.org/Part:BBa_K678046">pFun</a> has two NotI restriction sites flanking the device to be inserted. Hereby, the device can be cut out of the plasmid and the linearized DNA fragments can be transformed into the fungus. The devices for proof of concept are not <ins class="diffchange diffchange-inline">designed </ins>to be inserted at a specific site in the fungal genome. Therefore, the device can be integrated at any site and with a random number of copies by non-homologous end joining (NHEJ). This means that the possibility of the disruption of essential genes exists.<br><br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The fungal proof of concept plasmids, were constructed with the strong constitutive promoter <a href="http://partsregistry.org/Part:BBa_K678024">P<i>gpdA</i></a>, the <a href="http://partsregistry.org/Part:BBa_K678036">T<i>trpC</i></a> terminator, and the <a href="http://partsregistry.org/Part:BBa_K678040"><i>pyrG marker cassette</i></a>. Different genes encoding fluorescent proteins were included in the reporter system, where different compartments of fungi could be targeted. The reporter system was designed to target the nucleus, peroxisomes, and the mitochondria. All transformations were performed in the <i> Aspergillus nidulans</i> laboratory strain:<i> argB2, pyrG89, veA1</i>. </p> </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The fungal proof of concept plasmids, were constructed with the strong constitutive promoter <a href="http://partsregistry.org/Part:BBa_K678024">P<i>gpdA</i></a>, the <a href="http://partsregistry.org/Part:BBa_K678036">T<i>trpC</i></a> terminator, and the <a href="http://partsregistry.org/Part:BBa_K678040"><i>pyrG marker cassette</i></a>. Different genes encoding fluorescent proteins were included in the reporter system, where different compartments of fungi could be targeted. The reporter system was designed to target the nucleus, peroxisomes, and the mitochondria. All transformations were performed in the <i> Aspergillus nidulans</i> laboratory strain:<i> argB2, pyrG89, veA1</i>. </p> </div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>We <del class="diffchange diffchange-inline">have proved </del>that the Plug 'n' Play assembly standard can <del class="diffchange diffchange-inline">be </del>easily applied for the construction fungal plasmids. The constructed plasmids were transformed into <i>A. nidulans</i> and in each case the fluorescent protein was expressed. Only the strain transformed with <del class="diffchange diffchange-inline">the plasmid </del>targeting GFP to the mitochondria did not result in <del class="diffchange diffchange-inline">transform ants </del>and due to time limitation the experiment was not repeated.<br><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>We <ins class="diffchange diffchange-inline">here prove </ins>that the Plug 'n' Play assembly standard can easily <ins class="diffchange diffchange-inline">be </ins>applied for the construction fungal plasmids. The constructed plasmids were transformed into <i>A. nidulans</i> and in each case the fluorescent protein was expressed. Only the strain transformed with <ins class="diffchange diffchange-inline"><a href="http://partsregistry.org/Part:BBa_K678064">pJEJAM16</a> </ins>targeting GFP to the mitochondria did not result in <ins class="diffchange diffchange-inline">any transformants </ins>and due to time limitation the experiment was not repeated.<br><br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>The obtained transformants with the different constructed devices integrated into the genome all had a different morphology and were growing slower than the wild type strain. Especially the strain transformed with pJEJAM14 looked sick. The fungi also had a pronounced number of undefined vacuoles that can be <del class="diffchange diffchange-inline">seem </del>on the images obtained from differential interference contrast (DIC) microscopy. Such 'symptoms' are not uncommon when DNA is introduced into the genome by NHEJ. Below the images of the microscopy are presented.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>The obtained transformants with the different constructed devices integrated into the genome all had a different morphology and were growing slower than the wild type strain. Especially the strain transformed with pJEJAM14 looked sick. The fungi also had a pronounced number of undefined vacuoles that can be <ins class="diffchange diffchange-inline">seen </ins>on the images obtained from differential interference contrast (DIC) microscopy. Such 'symptoms' are not uncommon when DNA is introduced into the genome by NHEJ. Below the images of the microscopy are presented.</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Green fluorescence can be observed in the hyphae and in clear spots.This indicates that GFP is targeted to the peroxisomes, but further investigations are required to confirm this. The 'background' fluorescence was however not expected and a replication of the experiment should be conducted in order to be able to draw any conclusions upon this. </p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Green fluorescence can be observed in the hyphae and in clear spots. This indicates that GFP is targeted to the peroxisomes, but further investigations are required to confirm this. The 'background' fluorescence was however not expected and a replication of the experiment should be conducted in order to be able to draw any conclusions upon this. </p></div></td></tr>
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</table>Jaidehttp://2011.igem.org/wiki/index.php?title=Team:DTU-Denmark-2/results/Proofofconcept/fungi&diff=158715&oldid=prevJaide at 19:56, 21 September 20112011-09-21T19:56:12Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a name="Proof of concept"></a><h1><b>Proof of concept</b></h1></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a name="Proof of concept"></a><h1><b>Proof of concept</b></h1></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Five different plasmids were constructed with the Plug 'n' Play assembly standard in order to verify the systems function in filamentous fungi. To ensure a successful transformation the backbone plasmid <a href="http://partsregistry.org/Part:BBa_K678046">pFun</a> has two NotI restriction sites flanking the device to be inserted. Hereby, the device can be cut out of the plasmid and the <del class="diffchange diffchange-inline">linearised </del>DNA fragments can be transformed into the fungus. The devices for proof of concept are not design to be inserted at a specific site in the fungal genome. Therefore, the device can be integrated at any site and with a random number of copies by non homologous end-joining (NHEJ) in the fungus. This means that the possibility of the disruption of essential genes exists.<br><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Five different plasmids were constructed with the Plug 'n' Play assembly standard in order to verify the systems function in filamentous fungi. To ensure a successful transformation the backbone plasmid <a href="http://partsregistry.org/Part:BBa_K678046">pFun</a> has two NotI restriction sites flanking the device to be inserted. Hereby, the device can be cut out of the plasmid and the <ins class="diffchange diffchange-inline">linearized </ins>DNA fragments can be transformed into the fungus. The devices for proof of concept are not design to be inserted at a specific site in the fungal genome. Therefore, the device can be integrated at any site and with a random number of copies by non<ins class="diffchange diffchange-inline">-</ins>homologous end-joining (NHEJ) in the fungus. This means that the possibility of the disruption of essential genes exists.<br><br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The fungal proof of concept plasmids, were constructed with the strong constitutive promoter <a href="http://partsregistry.org/Part:BBa_K678024">P<i>gpdA</i></a>, the <a href="http://partsregistry.org/Part:BBa_K678036">T<i>trpC</i></a> terminator, and the <a href="http://partsregistry.org/Part:BBa_K678040"><i>pyrG marker cassette</i></a>. Different genes encoding fluorescent proteins were included in the reporter system, where different compartments of fungi could be targeted. The reporter system was designed to target the nucleus, peroxisomes, and the mitochondria. All transformations were performed in the <i> Aspergillus nidulans</i> laboratory strain:<i> argB2, pyrG89, veA1</i>. </p> </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The fungal proof of concept plasmids, were constructed with the strong constitutive promoter <a href="http://partsregistry.org/Part:BBa_K678024">P<i>gpdA</i></a>, the <a href="http://partsregistry.org/Part:BBa_K678036">T<i>trpC</i></a> terminator, and the <a href="http://partsregistry.org/Part:BBa_K678040"><i>pyrG marker cassette</i></a>. Different genes encoding fluorescent proteins were included in the reporter system, where different compartments of fungi could be targeted. The reporter system was designed to target the nucleus, peroxisomes, and the mitochondria. All transformations were performed in the <i> Aspergillus nidulans</i> laboratory strain:<i> argB2, pyrG89, veA1</i>. </p> </div></td></tr>
</table>Jaidehttp://2011.igem.org/wiki/index.php?title=Team:DTU-Denmark-2/results/Proofofconcept/fungi&diff=158671&oldid=prevJaide at 19:53, 21 September 20112011-09-21T19:53:19Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a name="Proof of concept"></a><h1><b>Proof of concept</b></h1></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a name="Proof of concept"></a><h1><b>Proof of concept</b></h1></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p align="justify"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p align="justify"></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Five different plasmids were constructed with the Plug 'n' Play assembly standard in order to verify the systems function in filamentous fungi. To ensure a successful transformation <del class="diffchange diffchange-inline">in fungi </del>the backbone plasmid <a href="http://partsregistry.org/Part:BBa_K678046">pFun</a> has two NotI restriction sites flanking the device to be inserted. Hereby, the device can be cut out of the plasmid and <del class="diffchange diffchange-inline">a </del>linearised DNA <del class="diffchange diffchange-inline">fragment </del>can be <del class="diffchange diffchange-inline">transformated </del>into the fungus. The devices for proof of concept <del class="diffchange diffchange-inline">in fungi </del>are not design to be inserted at a specific site in the fungal genome. Therefore, the device can be integrated at any site and with a random number of copies by non homologous end-joining (NHEJ) in the fungus. This means that the possibility of the disruption of essential genes exists.<br><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Five different plasmids were constructed with the Plug 'n' Play assembly standard in order to verify the systems function in filamentous fungi. To ensure a successful transformation the backbone plasmid <a href="http://partsregistry.org/Part:BBa_K678046">pFun</a> has two NotI restriction sites flanking the device to be inserted. Hereby, the device can be cut out of the plasmid and <ins class="diffchange diffchange-inline">the </ins>linearised DNA <ins class="diffchange diffchange-inline">fragments </ins>can be <ins class="diffchange diffchange-inline">transformed </ins>into the fungus. The devices for proof of concept are not design to be inserted at a specific site in the fungal genome. Therefore, the device can be integrated at any site and with a random number of copies by non homologous end-joining (NHEJ) in the fungus. This means that the possibility of the disruption of essential genes exists.<br><br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>The fungal proof of concept plasmids, were constructed with the <del class="diffchange diffchange-inline">same </del>strong constitutive promoter P<i>gpdA</i>, the T<i>trpC</i> terminator, and the <i>pyrG</i> <del class="diffchange diffchange-inline">marker cassette</del>. Different genes encoding fluorescent proteins were included in the reporter system, where different compartments of fungi could be targeted. The reporter system was designed to target the nucleus, peroxisomes, and the mitochondria. All transformations were performed in the <i> Aspergillus nidulans</i> laboratory strain:<i> argB2, pyrG89, veA1</i>. </p> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>The fungal proof of concept plasmids, were constructed with the strong constitutive promoter <ins class="diffchange diffchange-inline"><a href="http://partsregistry.org/Part:BBa_K678024"></ins>P<i>gpdA</i<ins class="diffchange diffchange-inline">></a</ins>>, the <ins class="diffchange diffchange-inline"><a href="http://partsregistry.org/Part:BBa_K678036"></ins>T<i>trpC</i<ins class="diffchange diffchange-inline">></a</ins>> terminator, and the <ins class="diffchange diffchange-inline"><a href="http://partsregistry.org/Part:BBa_K678040"></ins><i>pyrG <ins class="diffchange diffchange-inline">marker cassette</ins></i><ins class="diffchange diffchange-inline"></a></ins>. Different genes encoding fluorescent proteins were included in the reporter system, where different compartments of fungi could be targeted. The reporter system was designed to target the nucleus, peroxisomes, and the mitochondria. All transformations were performed in the <i> Aspergillus nidulans</i> laboratory strain:<i> argB2, pyrG89, veA1</i>. </p> </div></td></tr>
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</table>Jaidehttp://2011.igem.org/wiki/index.php?title=Team:DTU-Denmark-2/results/Proofofconcept/fungi&diff=158575&oldid=prevJaide at 19:47, 21 September 20112011-09-21T19:47:10Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a name="pJEJAM15 BBa_K678063"></a><h2>pJEJAM15 BBa_K678063</h2></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a name="pJEJAM15 BBa_K678063"></a><h2>pJEJAM15 BBa_K678063</h2></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p align="justify"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p align="justify"></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">Red fluorescence can be observed in clear spots. The occurrence of clear spots and compared to the results from device </del><a href="http://partsregistry.org/Part:<del class="diffchange diffchange-inline">BBa_K678062</del>"><del class="diffchange diffchange-inline">BBa_K678062</del></a><del class="diffchange diffchange-inline">, correlate with what expected </del>for <del class="diffchange diffchange-inline">the device </del><<del class="diffchange diffchange-inline">a href="http://partsregistry</del>.<del class="diffchange diffchange-inline">org/Part:BBa_K678063">BBa_K678063</del></<del class="diffchange diffchange-inline">a</del>> <del class="diffchange diffchange-inline">that holds </del>the gene <del class="diffchange diffchange-inline">for </del>red <del class="diffchange diffchange-inline">fluorescence </del>protein <del class="diffchange diffchange-inline">RFP </del>with <del class="diffchange diffchange-inline">the </del>targeting <del class="diffchange diffchange-inline">signal for </del>the <del class="diffchange diffchange-inline">nucleus. However</del>, <del class="diffchange diffchange-inline">we cannot conclude that </del>the <del class="diffchange diffchange-inline">signal is accumulated in </del>the <del class="diffchange diffchange-inline">nucleus, since they are not dyed</del>. <del class="diffchange diffchange-inline">Though, it </del>can be <del class="diffchange diffchange-inline">concluded </del>that the <del class="diffchange diffchange-inline">RFP signal </del>is <del class="diffchange diffchange-inline">targeting </del>to <del class="diffchange diffchange-inline">a specific place and accumulated somewhere in </del>the <del class="diffchange diffchange-inline">fungi compared </del>to <del class="diffchange diffchange-inline">the results from device <a href="http://partsregistry.org/Part:BBa_K678062">BBa_K678062</a></del>. </p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><a href="http://partsregistry.org/Part:<ins class="diffchange diffchange-inline">BBa_K678063</ins>"><ins class="diffchange diffchange-inline">pJEJAM15</ins></a> <ins class="diffchange diffchange-inline">is a plasmid intended </ins>for <ins class="diffchange diffchange-inline">linearization and transformation into </ins><<ins class="diffchange diffchange-inline">i>A</ins>. <ins class="diffchange diffchange-inline">nidulans</ins></<ins class="diffchange diffchange-inline">i</ins>><ins class="diffchange diffchange-inline">. The plasmid contains </ins>the gene <ins class="diffchange diffchange-inline">encoding monomeric </ins>red <ins class="diffchange diffchange-inline">fluorescent </ins>protein <ins class="diffchange diffchange-inline">(mRFP1) </ins>with <ins class="diffchange diffchange-inline">a nucleosomal </ins>targeting <ins class="diffchange diffchange-inline">sequence (NLS) fused to </ins>the <ins class="diffchange diffchange-inline">C-terminus</ins>, <ins class="diffchange diffchange-inline">under </ins>the <ins class="diffchange diffchange-inline">control of </ins>the <ins class="diffchange diffchange-inline">strong constitutive P<i>gpdA</i> promoter</ins>.<ins class="diffchange diffchange-inline"></p> Red fluorescence </ins>can be <ins class="diffchange diffchange-inline">observed in clear spots. To confirm </ins>that the <ins class="diffchange diffchange-inline">mRFP1 in fact </ins>is <ins class="diffchange diffchange-inline">targeted </ins>to the <ins class="diffchange diffchange-inline">nucleus further experiments have </ins>to <ins class="diffchange diffchange-inline">be conducted</ins>. </p></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"><p align="justify"></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">The constructed pJEJAM15 plasmid, holding device BBa_K678063, are cut at the two NotI site before transformation, ensuring linearised DNA fragment for optimal result.<br><br></p></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p align="center"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p align="center"></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><img src="https://static.igem.org/mediawiki/2011/e/e4/Device_25_ny.png" height="100px" align="center"> </img></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><img src="https://static.igem.org/mediawiki/2011/e/e4/Device_25_ny.png" height="100px" align="center"> </img></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><td>DIC image of<i>Aspergillus nidulans </i> with <br><a href="http://partsregistry.org/Part:BBa_K678063">pJEJAM15</a>.</td></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><td>DIC image of<i>Aspergillus nidulans </i> with <br><a href="http://partsregistry.org/Part:BBa_K678063">pJEJAM15</a>.</td></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><td>Fluorescence image of <i>Aspergillus nidulans </i> <del class="diffchange diffchange-inline">with device </del><br><a href="http://partsregistry.org/Part:BBa_K678063">pJEJAM15</a>.</td></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><td>Fluorescence image of <i>Aspergillus nidulans </i><br><a href="http://partsregistry.org/Part:BBa_K678063">pJEJAM15</a>.</td></div></td></tr>
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</table>Jaidehttp://2011.igem.org/wiki/index.php?title=Team:DTU-Denmark-2/results/Proofofconcept/fungi&diff=158422&oldid=prevMathilda at 19:38, 21 September 20112011-09-21T19:38:46Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><!--<span id="test">test</span>--></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><!--<span id="test">test</span>--></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#Proof of concept" class="h1">Proof of concept</a><br><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#Proof of concept" class="h1">Proof of concept</a><br><br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#pJEJAM12 BBa_K678060" class="h2"> pJEJAM12 BBa_K678060</a><br><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#pJEJAM12 BBa_K678060" class="h2"> pJEJAM12<ins class="diffchange diffchange-inline"><br></ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#pJEJAM13 BBa_K678061" class="h2"> pJEJAM13 BBa_K678061</a><br><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> </ins>BBa_K678060</a><br><br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#pJEJAM14 BBa_K678062" class="h2"> pJEJAM14 BBa_K678062</a><br><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#pJEJAM13 BBa_K678061" class="h2"> pJEJAM13 <ins class="diffchange diffchange-inline"><br></ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#pJEJAM15 BBa_K678063" class="h2"> pJEJAM15 BBa_K678063</a><br><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>BBa_K678061</a><br><br></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#pJEJAM14 BBa_K678062" class="h2"> pJEJAM14 <ins class="diffchange diffchange-inline"><br></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>BBa_K678062</a><br><br></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#pJEJAM15 BBa_K678063" class="h2"> pJEJAM15 <ins class="diffchange diffchange-inline"><br></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>BBa_K678063</a><br><br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#Wild type" class="h2"> Wild type</a><br><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#Wild type" class="h2"> Wild type</a><br><br></div></td></tr>
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</table>Mathildahttp://2011.igem.org/wiki/index.php?title=Team:DTU-Denmark-2/results/Proofofconcept/fungi&diff=158391&oldid=prevMathilda at 19:36, 21 September 20112011-09-21T19:36:37Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><!--<span id="test">test</span>--></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><!--<span id="test">test</span>--></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#Proof of concept" class="h1">Proof of concept</a><br><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#Proof of concept" class="h1">Proof of concept</a><br><br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#<del class="diffchange diffchange-inline">Device </del>BBa_K678060" class="h2"> pJEJAM12 BBa_K678060</a><br><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#<ins class="diffchange diffchange-inline">pJEJAM12 </ins>BBa_K678060" class="h2"> pJEJAM12 BBa_K678060</a><br><br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#<del class="diffchange diffchange-inline">Device </del>BBa_K678061" class="h2"> pJEJAM13 BBa_K678061</a><br><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#<ins class="diffchange diffchange-inline">pJEJAM13 </ins>BBa_K678061" class="h2"> pJEJAM13 BBa_K678061</a><br><br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#<del class="diffchange diffchange-inline">Device </del>BBa_K678062" class="h2"> pJEJAM14 BBa_K678062</a><br><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#<ins class="diffchange diffchange-inline">pJEJAM14 </ins>BBa_K678062" class="h2"> pJEJAM14 BBa_K678062</a><br><br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#<del class="diffchange diffchange-inline">Device </del>BBa_K678063" class="h2"> pJEJAM15 BBa_K678063</a><br><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#<ins class="diffchange diffchange-inline">pJEJAM15 </ins>BBa_K678063" class="h2"> pJEJAM15 BBa_K678063</a><br><br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#Wild type" class="h2"> Wild type</a><br><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a href="https://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#Wild type" class="h2"> Wild type</a><br><br></div></td></tr>
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</table>Mathildahttp://2011.igem.org/wiki/index.php?title=Team:DTU-Denmark-2/results/Proofofconcept/fungi&diff=158338&oldid=prevJaide at 19:33, 21 September 20112011-09-21T19:33:15Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p align="justify"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p align="justify"></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">Observed is red fluorescence spread evenly in the hyphae, which correlate with what expected for the device </del><a href="http://partsregistry.org/Part:BBa_K678062"><del class="diffchange diffchange-inline">BBa_K678062</del></a><del class="diffchange diffchange-inline">, that holds </del>the gene <del class="diffchange diffchange-inline">for </del>red <del class="diffchange diffchange-inline">fluorescence </del>protein <del class="diffchange diffchange-inline">RFP with no specific targeting signal</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><a href="http://partsregistry.org/Part:BBa_K678062"><ins class="diffchange diffchange-inline">pJEJAM14</ins></a> <ins class="diffchange diffchange-inline">is a plasmid intended for linearization and transformation into <i>A. nidulans</i>. The plasmid contains </ins>the gene <ins class="diffchange diffchange-inline">encoding monomeric </ins>red <ins class="diffchange diffchange-inline">fluorescent </ins>protein <ins class="diffchange diffchange-inline">(mRFP1) under the control of the strong constitutive P<i>gpdA</i> promoter. The expressed mRFP1 was evenly spread in the hyphae</ins>.</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"><p align="justify"></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">The constructed pJEJAM14 plasmid holding device BBa_K678062, are cut at the two NotI site before transformation, ensuring linearised DNA fragment for optimal result.<br><br></p></del></div></td><td colspan="2"> </td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><td>DIC image of <i>Aspergillus nidulans </i> with <del class="diffchange diffchange-inline">device </del><br><a href="http://partsregistry.org/Part:BBa_K678062"><del class="diffchange diffchange-inline">BBa_K678062</del></a> <del class="diffchange diffchange-inline">- detected with DIC light</del>.</td></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><td>DIC image of <i>Aspergillus nidulans </i> with <br><a href="http://partsregistry.org/Part:BBa_K678062"><ins class="diffchange diffchange-inline">pJEJAM14</ins></a>.</td></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><td>Fluorescence image of <i>Aspergillus nidulans </i> with <del class="diffchange diffchange-inline">device </del><br><a href="http://partsregistry.org/Part:BBa_K678062"><del class="diffchange diffchange-inline">BBa_K678062</del></a> <del class="diffchange diffchange-inline"> - detected with RFP filter</del>.</td></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><td>Fluorescence image of <i>Aspergillus nidulans </i> with <br><a href="http://partsregistry.org/Part:BBa_K678062"><ins class="diffchange diffchange-inline">pJEJAM14</ins></a>.</td></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></tr></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></tr></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></table></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></table></div></td></tr>
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<td colspan="2" class="diff-lineno">Line 216:</td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><tr></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><tr></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><td><i>Aspergillus nidulans </i> with <del class="diffchange diffchange-inline">device </del><br><a href="http://partsregistry.org/Part:BBa_K678062"<del class="diffchange diffchange-inline">BBa_K678062</del></a> - shown from front</td></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><td><i>Aspergillus nidulans </i> with <br><a href="http://partsregistry.org/Part:BBa_K678062"<ins class="diffchange diffchange-inline">>pJEJAM14</ins></a> - shown from front</td></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><td><i>Aspergillus nidulans </i> with <del class="diffchange diffchange-inline">device </del><br><a href="http://partsregistry.org/Part:BBa_K678062"<del class="diffchange diffchange-inline">BBa_K678062</del></a> - shown from back </td></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><td><i>Aspergillus nidulans </i> with <br><a href="http://partsregistry.org/Part:BBa_K678062"<ins class="diffchange diffchange-inline">>pJEJAM14</ins></a> - shown from back </td></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></tr></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></tr></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></table></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></table></div></td></tr>
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<td colspan="2" class="diff-lineno">Line 250:</td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></tr></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></tr></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><tr></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><tr></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><td>DIC image of<i>Aspergillus nidulans </i> with <del class="diffchange diffchange-inline">device </del><br><a href="http://partsregistry.org/Part:BBa_K678063"><del class="diffchange diffchange-inline">BBa_K678063</del></a> <del class="diffchange diffchange-inline">- detected with DIC light</del>.</td></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><td>DIC image of<i>Aspergillus nidulans </i> with <br><a href="http://partsregistry.org/Part:BBa_K678063"><ins class="diffchange diffchange-inline">pJEJAM15</ins></a>.</td></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><td>Fluorescence image of <i>Aspergillus nidulans </i> with device <br><a href="http://partsregistry.org/Part:BBa_K678063"><del class="diffchange diffchange-inline">BBa_K678063</del></a> <del class="diffchange diffchange-inline">- detected with RFP filter</del>.</td></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><td>Fluorescence image of <i>Aspergillus nidulans </i> with device <br><a href="http://partsregistry.org/Part:BBa_K678063"><ins class="diffchange diffchange-inline">pJEJAM15</ins></a>.</td></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></tr></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></tr></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></table></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></table></div></td></tr>
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<td colspan="2" class="diff-lineno">Line 262:</td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></tr></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></tr></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><tr></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><tr></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><td><i>Aspergillus nidulans </i> with <del class="diffchange diffchange-inline">device </del><br> <a href="http://partsregistry.org/Part:BBa_K678063"><del class="diffchange diffchange-inline">BBa_K678063</del></a> - Shown from front</td></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><td><i>Aspergillus nidulans </i> with <br> <a href="http://partsregistry.org/Part:BBa_K678063"><ins class="diffchange diffchange-inline">pJEJAM15</ins></a> - Shown from front</td></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><td><i>Aspergillus nidulans </i> with <del class="diffchange diffchange-inline">device </del><br> <a href="http://partsregistry.org/Part:BBa_K678063"><del class="diffchange diffchange-inline">BBa_K678063</del></a> - Shown from back </td></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><td><i>Aspergillus nidulans </i> with <br> <a href="http://partsregistry.org/Part:BBa_K678063"><ins class="diffchange diffchange-inline">pJEJAM15</ins></a> - Shown from back </td></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></tr></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></tr></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></table></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></table></div></td></tr>
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<td colspan="2" class="diff-lineno">Line 273:</td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a name="Wild type"></a><h2>Wild type</h2></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a name="Wild type"></a><h2>Wild type</h2></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>The <del class="diffchange diffchange-inline">control </del>strain shows no background or auto-fluorescence.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>The <ins class="diffchange diffchange-inline">wild type </ins>strain shows no background or auto-fluorescence.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p align="center"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p align="center"></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><table cellpadding="10px" align="center"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><table cellpadding="10px" align="center"></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><tr></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><tr></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><td> DIC image of <del class="diffchange diffchange-inline">Wild </del>type <i>Aspergillus nidulans </i>. <br></td></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><td> DIC image of <ins class="diffchange diffchange-inline">wild </ins>type <i>Aspergillus nidulans </i>. <br></td></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><td>RFP fluorescence image of wild type <i>Aspergillus nidulans </i>. <br> </td></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><td>RFP fluorescence image of wild type <i>Aspergillus nidulans </i>. <br> </td></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><td>GFP fluorescence image of wild type <i>Aspergillus nidulans </i>.<br></td></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><td>GFP fluorescence image of wild type <i>Aspergillus nidulans </i>.<br></td></div></td></tr>
</table>Jaidehttp://2011.igem.org/wiki/index.php?title=Team:DTU-Denmark-2/results/Proofofconcept/fungi&diff=158208&oldid=prevJaide at 19:25, 21 September 20112011-09-21T19:25:32Z<p></p>
<table style="background-color: white; color:black;">
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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 19:25, 21 September 2011</td>
</tr><tr><td colspan="2" class="diff-lineno">Line 142:</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a name="pJEJAM13 BBa_K678061"></a><h2>pJEJAM13 BBa_K678061</h2></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a name="pJEJAM13 BBa_K678061"></a><h2>pJEJAM13 BBa_K678061</h2></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p align="justify"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p align="justify"></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">Green fluorescence is can be observed in the hyphae and in clear spots. The occurrence of clear spots was expected for device </del><a href="http://partsregistry.org/Part:BBa_K678061"><del class="diffchange diffchange-inline">BBa_K678061</del></a><del class="diffchange diffchange-inline">, which holds the gene GFP encoding green fluorescence proteins </del>and <del class="diffchange diffchange-inline">targeting signal for the peroxiomes. This correlates fine with comparison of device BBa_K678060, which have non-targeting signal</del>. <<del class="diffchange diffchange-inline">br</del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><a href="http://partsregistry.org/Part:BBa_K678061"><ins class="diffchange diffchange-inline">pJEJAM13</ins></a> <ins class="diffchange diffchange-inline">is a plasmid intended for linearization </ins>and <ins class="diffchange diffchange-inline">transformation into <i>A</ins>. <ins class="diffchange diffchange-inline">nidulans</ins><<ins class="diffchange diffchange-inline">/i</ins>>. <ins class="diffchange diffchange-inline">The plasmid contains </ins>the <ins class="diffchange diffchange-inline">gene encoding green fluorescent protein (</ins>GFP<ins class="diffchange diffchange-inline">) with the peroxisomal </ins>targeting <ins class="diffchange diffchange-inline">sequence 1 (PTS1) fused </ins>to the <ins class="diffchange diffchange-inline">C-terminus, under </ins>the <ins class="diffchange diffchange-inline">control of the strong constitutive P</ins><<ins class="diffchange diffchange-inline">i</ins>><ins class="diffchange diffchange-inline">gpdA</ins></<ins class="diffchange diffchange-inline">i</ins>> <ins class="diffchange diffchange-inline">promoter</ins>.</p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">However, we cannot conclude that the signal is accumulated in the peroxisomes, since they are not dyed</del>. <del class="diffchange diffchange-inline">Though, it can be concluded that </del>the GFP <del class="diffchange diffchange-inline">signal is </del>targeting to <del class="diffchange diffchange-inline">a specific place and accumulated somewhere in </del>the <del class="diffchange diffchange-inline">fungi compared to </del>the <del class="diffchange diffchange-inline">results from device </del><<del class="diffchange diffchange-inline">a href="http://partsregistry.org/Part:BBa_K678060"</del>><del class="diffchange diffchange-inline">BBa_K678060</del></<del class="diffchange diffchange-inline">a</del>>. </p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"><p align="justify"></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">The plasmid constructed pJEJAM13 holding device BBa_K678061, are cut at the two NotI site before transformation, ensuring linearised DNA fragment for optimal result.<br><br></p></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p align="center"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p align="center"></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><img src="https://static.igem.org/mediawiki/2011/6/68/Device_23_ny.png" height="100px" align="center"> </img></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><img src="https://static.igem.org/mediawiki/2011/6/68/Device_23_ny.png" height="100px" align="center"> </img></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"><br></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"><br></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"><p align="justify"></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">Green fluorescence can be observed in the hyphae and in clear spots.This indicates that GFP is targeted to the peroxisomes, but further investigations are required to confirm this. The 'background' fluorescence was however not expected and a replication of the experiment should be conducted in order to be able to draw any conclusions upon this. </p></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><tr></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><tr></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><td>DIC image of <i>Aspergillus nidulans </i> with <del class="diffchange diffchange-inline">device </del><br><a href="http://partsregistry.org/Part:BBa_K678061">pJEJAM13</a> <del class="diffchange diffchange-inline">- detected with DIC light</del>.</td></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><td>DIC image of <i>Aspergillus nidulans </i> with <br><a href="http://partsregistry.org/Part:BBa_K678061">pJEJAM13</a>.</td></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><td>Fluorescence image of <i>Aspergillus nidulans </i> with <br><a href="http://partsregistry.org/Part:BBa_K678061">pJEJAM13</a>. Green fluorescence can be observed in the hyphae and in clear spots.</td></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><td>Fluorescence image of <i>Aspergillus nidulans </i> with <br><a href="http://partsregistry.org/Part:BBa_K678061">pJEJAM13</a>. Green fluorescence can be observed in the hyphae and in clear spots.</td></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></tr></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></tr></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></tr></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></tr></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><tr></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><tr></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><td><i>Aspergillus nidulans </i> with <del class="diffchange diffchange-inline">device </del><br><a href="http://partsregistry.org/Part:BBa_K678061"><del class="diffchange diffchange-inline">BBa_K678061</del></a> - Shown from front</td></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><td><i>Aspergillus nidulans </i> with <br><a href="http://partsregistry.org/Part:BBa_K678061"><ins class="diffchange diffchange-inline">pJEJAM13</ins></a> - Shown from front</td></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><td><i>Aspergillus nidulans </i> with <del class="diffchange diffchange-inline">device </del><br><a href="http://partsregistry.org/Part:BBa_K678061"><del class="diffchange diffchange-inline">BBa_K678061</del></a> - Shown from back </td></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><td><i>Aspergillus nidulans </i> with <br><a href="http://partsregistry.org/Part:BBa_K678061"><ins class="diffchange diffchange-inline">pJEJAM13</ins></a> - Shown from back </td></div></td></tr>
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</table>Jaidehttp://2011.igem.org/wiki/index.php?title=Team:DTU-Denmark-2/results/Proofofconcept/fungi&diff=158053&oldid=prevJaide at 19:14, 21 September 20112011-09-21T19:14:40Z<p></p>
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<td colspan='2' style="background-color: white; color:black;">Revision as of 19:14, 21 September 2011</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a name="pJEJAM12 BBa_K678060"></a><h2>pJEJAM12 BBa_K678060</h2></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a name="pJEJAM12 BBa_K678060"></a><h2>pJEJAM12 BBa_K678060</h2></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p align="justify"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p align="justify"></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>href="http://partsregistry.org/Part:BBa_K678060">pJEJAM12</a> is a plasmid intended for linearization and transformation into <i>A. nidulans</i>. The plasmid contains the gene encoding green fluorescent protein (GFP) under the control of the strong constitutive P<i>gpdA</i> promoter. The expressed GFP was evenly spread in the hyphae.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><a </ins>href="http://partsregistry.org/Part:BBa_K678060">pJEJAM12</a> is a plasmid intended for linearization and transformation into <i>A. nidulans</i>. The plasmid contains the gene encoding green fluorescent protein (GFP) under the control of the strong constitutive P<i>gpdA</i> promoter. The expressed GFP was evenly spread in the hyphae.</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><td><i>Aspergillus nidulans </i> with <del class="diffchange diffchange-inline">device </del><br><a href="http://partsregistry.org/Part:BBa_K678060"><del class="diffchange diffchange-inline">BBa_K678060</del></a> - Shown from front</td></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><td><i>Aspergillus nidulans </i> with <br><a href="http://partsregistry.org/Part:BBa_K678060"><ins class="diffchange diffchange-inline">pJEJAM12</ins></a> - Shown from front</td></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><td><i>Aspergillus nidulans </i> with <del class="diffchange diffchange-inline">device </del><br><a href="http://partsregistry.org/Part:BBa_K678060"><del class="diffchange diffchange-inline">BBa_K678060</del></a> - Shown from Back </td></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><td><i>Aspergillus nidulans </i> with <br><a href="http://partsregistry.org/Part:BBa_K678060"><ins class="diffchange diffchange-inline">pJEJAM12</ins></a> - Shown from Back </td></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><td>DIC image of <i>Aspergillus nidulans </i> with device <br><a href="http://partsregistry.org/Part:BBa_K678061"><del class="diffchange diffchange-inline">BBa_K678061</del></a> - detected with DIC light.</td></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><td>DIC image of <i>Aspergillus nidulans </i> with device <br><a href="http://partsregistry.org/Part:BBa_K678061"><ins class="diffchange diffchange-inline">pJEJAM13</ins></a> - detected with DIC light.</td></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><td>Fluorescence image of <i>Aspergillus nidulans </i> with <del class="diffchange diffchange-inline">device </del><br><a href="http://partsregistry.org/Part:BBa_K678061"><del class="diffchange diffchange-inline">BBa_K678061</del></a> <del class="diffchange diffchange-inline">- detected with GFP filter</del>.</td></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><td>Fluorescence image of <i>Aspergillus nidulans </i> with <br><a href="http://partsregistry.org/Part:BBa_K678061"><ins class="diffchange diffchange-inline">pJEJAM13</ins></a><ins class="diffchange diffchange-inline">. Green fluorescence can be observed in the hyphae and in clear spots</ins>.</td></div></td></tr>
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</table>Jaidehttp://2011.igem.org/wiki/index.php?title=Team:DTU-Denmark-2/results/Proofofconcept/fungi&diff=157989&oldid=prevJaide at 19:10, 21 September 20112011-09-21T19:10:47Z<p></p>
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<td colspan='2' style="background-color: white; color:black;">Revision as of 19:10, 21 September 2011</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p align="justify"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p align="justify"></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">Green fluorescence can be observed evenly spread in the hyphae. This correlate with what was expected for the device <a </del>href="http://partsregistry.org/Part:BBa_K678060"><del class="diffchange diffchange-inline">BBa_K678060</del></a> <del class="diffchange diffchange-inline">that holds the gene </del><i><del class="diffchange diffchange-inline">GFP</del></i> <del class="diffchange diffchange-inline">for green fluorescence and with no specific targeting signal</del>. </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>href="http://partsregistry.org/Part:BBa_K678060"><ins class="diffchange diffchange-inline">pJEJAM12</ins></a> <ins class="diffchange diffchange-inline">is a plasmid intended for linearization and transformation into </ins><i><ins class="diffchange diffchange-inline">A. nidulans</ins></i>. <ins class="diffchange diffchange-inline">The plasmid contains the gene encoding green fluorescent protein (GFP) under the control of the strong constitutive P</ins><<ins class="diffchange diffchange-inline">i</ins>><ins class="diffchange diffchange-inline">gpdA</ins><<ins class="diffchange diffchange-inline">/i</ins>> <ins class="diffchange diffchange-inline">promoter. The expressed GFP was evenly spread in the hyphae.</ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><<del class="diffchange diffchange-inline">/p</del>> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><<del class="diffchange diffchange-inline">br</del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p <del class="diffchange diffchange-inline">align="justify"</del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><<ins class="diffchange diffchange-inline">/</ins>p> </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">The constructed pJEJAM12 plasmid consist of device BBa_K678060 (<i>pgdA,GFP,TrpC,pyrG</i>) , which are cut at the two NotI site before transformation, ensuring linearised DNA fragment for optimal result.</del><br<del class="diffchange diffchange-inline">><br></p</del>> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><br> </div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><img src="https://static.igem.org/mediawiki/2011/e/ef/Device_22_ny.png" height="100px" align="center"> </img></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><img src="https://static.igem.org/mediawiki/2011/e/ef/Device_22_ny.png" height="100px" align="center"> </img></div></td></tr>
</table>Jaide