Difference between revisions of "Team:Utrecht/Parts"
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<div class = "customelementM5B" id = "PnT"> | <div class = "customelementM5B" id = "PnT"> | ||
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<table class="InterlabTable" width = "100%"> | <table class="InterlabTable" width = "100%"> | ||
<caption style = "font-size: 1.5em;background-color: #A4C2F4">Table 1: Used Biobricks</caption> | <caption style = "font-size: 1.5em;background-color: #A4C2F4">Table 1: Used Biobricks</caption> | ||
<tr><th>Name</th><th>Biobrick Number</th><th>Size</th><th>Size</th><th>Obtained</th></tr> | <tr><th>Name</th><th>Biobrick Number</th><th>Size</th><th>Size</th><th>Obtained</th></tr> | ||
| − | <tr><td>RFP</td><td>BBa_K516032</td><td>828</td><td>Red fluorescence</td><td>isolated</ | + | <tr><td>RFP</td><td>BBa_K516032</td><td>828</td><td>Red 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>isolated</ | + | <tr><td>Promoter</td><td>BBa_K608003</td><td>56 </td><td>Strong promoter with medium ribosome binding site</td><td>isolated</td></tr> |
| − | <tr><td>eYFP</td><td>BBa_I15017</td><td>724</td><td>Yellow fluorescence</td><td>isolated</ | + | <tr><td>eYFP</td><td>BBa_I15017</td><td>724</td><td>Yellow fluorescence</td><td>isolated</td></tr> |
| − | <tr><td>CheZ</td><td>BBa_K629003</td><td>645</td><td>CheZ chemotaxis regulator protein phosphatase for CheY</td><td>isolated</ | + | <tr><td>CheZ</td><td>BBa_K629003</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</ | + | <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>BBa_K2736006</td><td>1505</td><td>Fusion protein of CusS with the Tar ligand binding domain</td><td> ordered</ | + | <tr><td>Custom receptor</td><td>BBa_K2736006</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_K2736002</td><td>1655</td><td>CheZ fused to Rluc</td><td>ordered</ | + | <tr><td>1/5A::CheZ::Rluc</td><td>BBa_K2736002</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</ | + | <tr><td>CheY</td><td>-</td><td>390</td><td>Chemotaxis response regulator protein</td><td>ordered</td></tr> |
</table> | </table> | ||
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</div> | </div> | ||
<div class = "customelementM5A" id = "Usage"> | <div class = "customelementM5A" id = "Usage"> | ||
| − | < | + | |
| − | < | + | <table class="InterlabTable" width = "100%"> |
| − | + | <caption style = "font-size: 1.5em;background-color: #A4C2F4">Table 1: Used Biobricks</caption> | |
| − | </ | + | <tr><th>Name</th><th>Biobrick Number</th><th>Size</th><th>Size</th><th>Obtained</th></tr> |
| + | <tr><td>1/5A::eYFP::CheY</td><td>BBa_K2736000</td><td>1865</td><td>Chemotaxis responsive BRET pair</td><td>Created | ||
| + | <tr><td>1/5A::CheZ::Rluc </td><td>BBa_K2736001</td><td>930</td><td>RFP behind a promoter that responds to CusR</td><td>Created</td></tr> | ||
| + | <tr><td>Cu_prom::RFP</td><td>BBa_K2736002</td><td>1655</td><td>Chemotaxis responsive BRET pair</td><td>Created | ||
| + | <tr><td>Tar(E491A)</td><td>BBa_K2736003</td><td>1703</td><td>Tar receptor protein with a E->A substitution</td><td>Created</td></tr> | ||
| + | <tr><td>Tar(Q309A)</td><td>BBa_K2736004</td><td>1703</td><td>Tar receptor protein with a Q->A substitution Created</td></tr> | ||
| + | <tr><td>Tar(E491A, Q309A)</td><td>BBa_K2736005</td><td>1703</td><td>Tar receptor with two amino acid mutations Created</td></tr> | ||
| + | </table> | ||
| + | |||
</div> | </div> | ||
Revision as of 11:41, 9 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_K2736000 and BBa_K2736001, were created to be used as BRET pair to measure the activity of our customized Tar receptors (BBa_K2736003, BBa_K2736004, and BBa_K2736005). To test whether it was possible to switch ligand binding domains of the Tar receptor and two component receptors we created BBa_K2736002, and BBa_K2736006.
| Name | Biobrick Number | Size | Size | Obtained |
|---|---|---|---|---|
| RFP | BBa_K516032 | 828 | Red fluorescence | isolated |
| Promoter | BBa_K608003 | 56 | Strong promoter with medium ribosome binding site | isolated |
| eYFP | BBa_I15017 | 724 | Yellow fluorescence | isolated |
| CheZ | BBa_K629003 | 645 | CheZ chemotaxis regulator protein phosphatase for CheY | isolated |
| Copper promoter | - | 98 | Promoter responsive to phosphorylated CusR | ordered |
| Custom receptor | BBa_K2736006 | 1505 | Fusion protein of CusS with the Tar ligand binding domain | ordered |
| 1/5A::CheZ::Rluc | BBa_K2736002 | 1655 | CheZ fused to Rluc | ordered |
| CheY | - | 390 | Chemotaxis response regulator protein | ordered |
| Name | Biobrick Number | Size | Size | Obtained |
|---|---|---|---|---|
| 1/5A::eYFP::CheY | BBa_K2736000 | 1865 | Chemotaxis responsive BRET pair | Created |
| 1/5A::CheZ::Rluc | BBa_K2736001 | 930 | RFP behind a promoter that responds to CusR | Created |
| Cu_prom::RFP | BBa_K2736002 | 1655 | Chemotaxis responsive BRET pair | Created |
| Tar(E491A) | BBa_K2736003 | 1703 | Tar receptor protein with a E->A substitution | Created |
| Tar(Q309A) | BBa_K2736004 | 1703 | Tar receptor protein with a Q->A substitution Created | |
| Tar(E491A, Q309A) | BBa_K2736005 | 1703 | Tar receptor with two amino acid mutations Created |