Difference between revisions of "Team:Utrecht/Parts"
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<div class = "sidenav" id = "side-menu"> | <div class = "sidenav" id = "side-menu"> | ||
<ul> | <ul> | ||
| − | <li><a href="#Introduction"></a></li> | + | <li><a href="#Introduction">Introduction</a></li> |
| − | <li><a href="# | + | <li><a href="#BBU">Parts Acquired</a></li> |
| − | <li><a href="# | + | <li><a href="#BBC">Parts Created</a></li> |
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</ul> | </ul> | ||
</div> | </div> | ||
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<div class = "customelementM5A" id = "Introduction"> | <div class = "customelementM5A" id = "Introduction"> | ||
| − | <p>Several biobricks from the distribution kit were used during our project. These were retrieved from the kit using the protocols provided by iGEM, transformed into <i>E. coli</i> | + | <p>Several biobricks from the distribution kit were used during our project. These were retrieved from the kit using the protocols provided by iGEM<a class = "url_icon" href = "http://parts.igem.org/Help:2018_DNA_Distribution"></a>, transformed into <i>E. coli</i> and isolated in elution buffer NE. We also ordered several constructs from IDT (Table 1). These Biobricks were used to create several new biobricks that were relevant to our project. BBa_K2736101<a class = "url_icon" href = "http://parts.igem.org/Part:BBa_K2736101"></a> and BBa_K2736103<a class = "url_icon" href = "http://parts.igem.org/Part:BBa_K2736103"></a>, were created to be used as BRET pair to measure the activity of our customized Tar receptors (BBa_K2736106<a class = "url_icon" href = "http://parts.igem.org/Part:BBa_K2736106"></a>, BBa_K2736108<a class = "url_icon" href = "http://parts.igem.org/Part:BBa_K2736108"></a>). To test whether it was possible to switch ligand binding domains of the Tar receptor and two component receptors we created BBa_K2736105<a class = "url_icon" href = "http://parts.igem.org/Part:BBa_K2736105"></a>, and BBa_K2736109<a class = "url_icon" href = "http://parts.igem.org/Part:BBa_K2736109"></a>. <b>During our project we proved the functionallity of the parts BBa_K2736101<a class = "url_icon" href = "http://parts.igem.org/Part:BBa_K2736101"></a> and BBa_K2736105<a class = "url_icon" href = "http://parts.igem.org/Part:BBa_K2736105"></a>, which were submitted to the registery.</b></p> |
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| − | Tar receptors | + | |
| − | switch ligand binding domains of the Tar receptor and two component receptors we created | + | |
| − | + | ||
</div> | </div> | ||
| − | <div class = "customelementM5B" id = " | + | <div class = "customelementM5B" id = "BBU"> |
| − | + | <table class="InterlabTable" width = "100%"> | |
| − | + | <caption style = "font-size: 1.5em;background-color: #A4C2F4">Table 1: Acquired Biobricks</caption> | |
| − | <table class="InterlabTable"> | + | <tr><th>Name</th><th>Biobrick Number</th><th>Size</th><th>Function</th><th>Obtained</th></tr> |
| − | <caption style = "font-size: 1.5em;background-color: #A4C2F4">Table 1: | + | <tr><td>RFP</td><td>BBa_K516032<a class = "url_icon" href = "http://parts.igem.org/Part:BBa_K516032"></a></td><td>828</td><td>Red fluorescence</td><td>Isolated</td></tr> |
| − | <tr><th>Name</th><th>Biobrick Number</th><th>Size</th><th> | + | <tr><td>Promoter</td><td>BBa_K608003<a class = "url_icon" href = "http://parts.igem.org/Part:BBa_K608003"></a></td><td>56</td><td>Strong promoter with medium ribosome binding site</td><td>Isolated</td></tr> |
| − | <tr><td>RFP</td><td>BBa_K516032</td><td>828</td><td>Red fluorescence</td><td> | + | <tr><td>eYFP</td><td>BBa_I15017<a class = "url_icon" href = "http://parts.igem.org/Part:BBa_I15017"></a></td><td>724</td><td>Yellow fluorescence</td><td>Isolated</td></tr> |
| − | <tr><td>Promoter</td><td>BBa_K608003</td><td>56 </td><td>Strong promoter with medium ribosome binding site</td><td> | + | <tr><td>CheZ</td><td>BBa_K629003<a class = "url_icon" href = "http://parts.igem.org/Part:BBa_K629003"></a></td><td>645</td><td>CheZ chemotaxis regulator protein phosphatase for CheY</td><td>Isolated</td></tr> |
| − | <tr><td>eYFP</td><td>BBa_I15017</td><td>724</td><td>Yellow fluorescence</td><td> | + | <tr><td>Copper promoter</td><td>-</td><td>98</td><td>Promoter responsive to phosphorylated CusR </td><td>Ordered</td></tr> |
| − | <tr><td>CheZ</td><td>BBa_K629003</td><td>645</td><td> | + | <tr><td>Custom receptor</td><td>BBa_K2736109<a class = "url_icon" href = "http://parts.igem.org/Part:BBa_K2736109"></a></td><td>1505</td><td>Fusion protein of CusS with the Tar ligand binding domain</td><td> Ordered</td></tr> |
| − | <tr><td>Copper promoter</td><td>98</td><td>Promoter responsive to phosphorylated CusR </td><td> | + | <tr><td>1/5A::CheZ::Rluc</td><td>BBa_K2736103<a class = "url_icon" href = "http://parts.igem.org/Part:BBa_K2736103"></a></td><td>1655</td><td>CheZ fused to Rluc</td><td>Ordered</td></tr> |
| − | <tr><td>Custom receptor</td><td> | + | <tr><td>CheY</td><td>-</td><td>390</td><td>Chemotaxis response regulator protein</td><td>Ordered</td></tr> |
| − | <tr><td>1/5A::CheZ::Rluc</td><td> | + | |
| − | <tr><td>CheY</td><td>390</td><td>Chemotaxis response regulator protein </td><td> | + | |
</table> | </table> | ||
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</div> | </div> | ||
| − | <div class = "customelementM5A" id = " | + | <div class = "customelementM5A" id = "BBC"> |
| − | < | + | |
| − | < | + | <table class="InterlabTable" width = "100%"> |
| − | + | <caption style = "font-size: 1.5em;background-color: #A4C2F4">Table 2: Created Biobricks</caption> | |
| − | </ | + | <tr><th>Name</th><th>Biobrick Number</th><th>Size</th><th>Function</th><th>Obtained</th></tr> |
| + | <tr><td>1/5A::eYFP::CheY</td><td>BBa_K2736101<a class = "url_icon" href = "http://parts.igem.org/Part:BBa_K2736101"></a></td><td>1865</td><td>Chemotaxis responsive BRET pair (Part 1)</td><td>Created</td></tr> | ||
| + | <tr><td>1/5A::CheZ::Rluc </td><td>BBa_K2736103<a class = "url_icon" href = "http://parts.igem.org/Part:BBa_K2736103"></a></td><td>1655</td><td>Chemotaxis responsive BRET pair (Part 2)</td><td>Created</td></tr> | ||
| + | <tr><td>Cu_prom::RFP</td><td>BBa_K2736105<a class = "url_icon" href = "http://parts.igem.org/Part:BBa_K2736105"></a></td><td>930</td><td>RFP behind a promoter that responds to CusR</td><td>Created</td></tr> | ||
| + | <tr><td>Tar(E491A)</td><td>BBa_K2736106<a class = "url_icon" href = "http://parts.igem.org/Part:BBa_K2736106"></a></td><td>1703</td><td>Tar receptor protein with a E->A substitution</td><td>Created</td></tr> | ||
| + | </table> | ||
| + | |||
</div> | </div> | ||
| + | <div class = "navigateArrows"> | ||
| + | <div class = "arrowLeft"> <a href="https://2018.igem.org/Team:Utrecht/Design"><b class = "fa fa-angle-left"></b> Project Design</a></div> | ||
| + | <div class = "arrowRight"> <a href="https://2018.igem.org/Team:Utrecht/Improve">Improved Biobrick <b class = "fa fa-angle-right"></b></a></div> | ||
| + | </div> | ||
</body> | </body> | ||
Latest revision as of 21:12, 4 December 2018
Several biobricks from the distribution kit were used during our project. These were retrieved from the kit using the protocols provided by iGEM, transformed into E. coli and isolated in elution buffer NE. We also ordered several constructs from IDT (Table 1). These Biobricks were used to create several new biobricks that were relevant to our project. BBa_K2736101 and BBa_K2736103, were created to be used as BRET pair to measure the activity of our customized Tar receptors (BBa_K2736106, BBa_K2736108). To test whether it was possible to switch ligand binding domains of the Tar receptor and two component receptors we created BBa_K2736105, and BBa_K2736109. During our project we proved the functionallity of the parts BBa_K2736101 and BBa_K2736105, which were submitted to the registery.