Difference between revisions of "Team:Utrecht/Parts"

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<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>1/5A::eYFP::CheY</td><td>BBa_K2736000</td><td>1865</td><td>Chemotaxis responsive BRET pair (Part 1)</td><td>Created</td></tr>
 
<tr><td>1/5A::eYFP::CheY</td><td>BBa_K2736000</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_K2736001</td><td>930</td><td>Chemotaxis responsive BRET pair (Part 2)</td><td>Created</td></tr>
+
<tr><td>1/5A::CheZ::Rluc </td><td>BBa_K2736001</td><td>1655</td><td>Chemotaxis responsive BRET pair (Part 2)</td><td>Created</td></tr>
<tr><td>Cu_prom::RFP</td><td>BBa_K2736002</td><td>1655</td><td>RFP behind a promoter that responds to CusR</td><td>Created</td></tr>
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<tr><td>Cu_prom::RFP</td><td>BBa_K2736002</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_K2736003</td><td>1703</td><td>Tar receptor protein with a E->A substitution</td><td>Created</td></tr>
 
<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</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</td><td>Created</td></tr>

Revision as of 12:33, 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.

Table 1: Acquired Biobricks
NameBiobrick NumberSizeSizeObtained
RFPBBa_K516032828Red fluorescenceIsolated
PromoterBBa_K60800356Strong promoter with medium ribosome binding siteIsolated
eYFPBBa_I15017724Yellow fluorescenceIsolated
CheZBBa_K629003645CheZ chemotaxis regulator protein phosphatase for CheYIsolated
Copper promoter-98Promoter responsive to phosphorylated CusR Ordered
Custom receptorBBa_K27360061505Fusion protein of CusS with the Tar ligand binding domain Ordered
1/5A::CheZ::RlucBBa_K27360011655CheZ fused to RlucOrdered
CheY-390Chemotaxis response regulator proteinOrdered
Table 2: Created Biobricks
NameBiobrick NumberSizeSizeObtained
1/5A::eYFP::CheYBBa_K27360001865Chemotaxis responsive BRET pair (Part 1)Created
1/5A::CheZ::Rluc BBa_K27360011655Chemotaxis responsive BRET pair (Part 2)Created
Cu_prom::RFPBBa_K2736002930RFP behind a promoter that responds to CusRCreated
Tar(E491A)BBa_K27360031703Tar receptor protein with a E->A substitutionCreated
Tar(Q309A)BBa_K27360041703Tar receptor protein with a Q->A substitutionCreated
Tar(E491A, Q309A)BBa_K27360051703Tar receptor with two amino acid mutationsCreated