Difference between revisions of "Team:Utrecht/Parts"
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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<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 | + | <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>.</p> |
</div> | </div> | ||
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<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>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>Copper promoter</td><td>-</td><td>98</td><td>Promoter responsive to phosphorylated CusR </td><td>Ordered</td></tr> | <tr><td>Copper promoter</td><td>-</td><td>98</td><td>Promoter responsive to phosphorylated CusR </td><td>Ordered</td></tr> | ||
| − | <tr><td>Custom receptor</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>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>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>CheY</td><td>-</td><td>390</td><td>Chemotaxis response regulator protein</td><td>Ordered</td></tr> | <tr><td>CheY</td><td>-</td><td>390</td><td>Chemotaxis response regulator protein</td><td>Ordered</td></tr> | ||
Revision as of 19:46, 17 October 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.
| Name | Biobrick Number | Size | Function | Obtained |
|---|---|---|---|---|
| 1/5A::eYFP::CheY | BBa_K2736101 | 1865 | Chemotaxis responsive BRET pair (Part 1) | Created |
| 1/5A::CheZ::Rluc | BBa_K2736103 | 1655 | Chemotaxis responsive BRET pair (Part 2) | Created |
| Cu_prom::RFP | BBa_K2736105 | 930 | RFP behind a promoter that responds to CusR | Created |
| Tar(E491A) | BBa_K2736106 | 1703 | Tar receptor protein with a E->A substitution | Created |