Difference between revisions of "Team:Utrecht/Parts"

Revision as of 10:10, 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, 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.

Used Biobricks

Table 1: Used Biobricks
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

Created Biobricks

Here comes Table 2

@@ Line 31: / Line 31: @@
 <div class = "customelementM5A" id = "Introduction">
-<p>Several biobricks from the kit were used during our project. Some of these were isolated from the kit by
+<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>, 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.
-direct transformation of 2 μl of DNA resuspended in elution buffer NE. Besides the biobricks that were
+BBa_K2736000 and BBa_K2736001, were created to be used as BRET pair to measure the activity of our customized
-isolated from the kit, some constructs were ordered completely 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 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
@@ Line 51: / Line 48: @@
 <tr><td>Promoter</td><td>BBa_K608003</td><td>56	</td><td>Strong promoter with medium ribosome binding site</td><td>isolated</th></tr>
 <tr><td>eYFP</td><td>BBa_I15017</td><td>724</td><td>Yellow fluorescence</td><td>isolated</th></tr>
-<tr><td>CheZ</td><td>BBa_K629003</td><td>645	</td><td>"cheZ  chemotaxis regulator  protein phosphatase for CheY"</td><td>isolated</th></tr>
+<tr><td>CheZ</td><td>BBa_K629003</td><td>645</td><td>"cheZ  chemotaxis regulator  protein phosphatase for CheY"</td><td>isolated</th></tr>
 <tr><td>Copper promoter</td><td>98</td><td>Promoter responsive to phosphorylated CusR	</td><td>ordered</th></tr>
-<tr><td>Custom receptor</td><td>BBa_K2736006</td><td>1505	Fusion protein of CusS with the Tar ligand binding domain</td><td>	ordered</th></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</th></tr>
-<tr><td>1/5A::CheZ::Rluc</td><td>BBa_K2736002</td><td>1655CheZ fused to Rluc</td><td>ordered</th></tr>
+<tr><td>1/5A::CheZ::Rluc</td><td>BBa_K2736002</td><td>1655</td><td>CheZ fused to Rluc</td><td>ordered</th></tr>
 <tr><td>CheY</td><td>390</td><td>Chemotaxis response regulator protein	</td><td>ordered</th></tr>
 </table>