Team:DTU-Denmark/Results

From 2010.igem.org

(Difference between revisions)
Line 139: Line 139:
     <td align="center" ><font face="arial" size="3"><a class="mainLinks" href="https://2010.igem.org/Team:DTU-Denmark/Project" >The Project</a> </font></td>
     <td align="center" ><font face="arial" size="3"><a class="mainLinks" href="https://2010.igem.org/Team:DTU-Denmark/Project" >The Project</a> </font></td>
     <td align="center" ><font face="arial" size="3"><a class="mainLinks" href="https://2010.igem.org/Team:DTU-Denmark/Parts" >Parts submitted</a> </font></td>
     <td align="center" ><font face="arial" size="3"><a class="mainLinks" href="https://2010.igem.org/Team:DTU-Denmark/Parts" >Parts submitted</a> </font></td>
-
     <td align="center" ><font face="arial" size="3"><a class="mainLinks" href="https://2010.igem.org/Team:DTU-Denmark/Modelling">Modelling</a></font> </td>
+
     <td align="center" ><font face="arial" size="3"><a class="mainLinks" href="https://2010.igem.org/Team:DTU-Denmark/Results">Results</a></font> </td>
     <td align="center" ><font face="arial" size="3"><a class="mainLinks" href="https://2010.igem.org/Team:DTU-Denmark/Notebook" title="Day to day lab activity">Notebook</a>  
     <td align="center" ><font face="arial" size="3"><a class="mainLinks" href="https://2010.igem.org/Team:DTU-Denmark/Notebook" title="Day to day lab activity">Notebook</a>  
   <td align="center" ><font face="arial" size="3"><a class="mainLinks" href="https://2010.igem.org/Team:DTU-Denmark/Blog">Blog</a></font> </td>
   <td align="center" ><font face="arial" size="3"><a class="mainLinks" href="https://2010.igem.org/Team:DTU-Denmark/Blog">Blog</a></font> </td>
Line 152: Line 152:
   <font color="#990000" face="arial" size="3">
   <font color="#990000" face="arial" size="3">
<ul type="circle">
<ul type="circle">
-
<li><a href="https://2010.igem.org/Team:DTU-Denmark/Experimental_Setup">Experimental Setup</a></li>
+
<li><a href="https://2010.igem.org/Team:DTU-Denmark/Repressor_Section">Repressor - Anti-Repressor Section</a></li>
<ul>
<ul>
<font size="2">
<font size="2">
-
<li><a href="https://2010.igem.org/Team:DTU-Denmark/Experimental_Setup_repressors" target="_blank">Repressor Section</a></li>
+
<li><a href="https://2010.igem.org/Team:DTU-Denmark/Repressor_Section#Experimental" target="_blank">Experimental Setup</a></li>
-
<li><a href="https://2010.igem.org/Team:DTU-Denmark/Experimental_Setup_antiterminators" target="_blank">Anti-Terminators Section</a></li>
+
<li><a href="https://2010.igem.org/Team:DTU-Denmark/Repressor_Section#Characterization" target="_blank">Characterization Results</a></li>
</ul><br></font>
</ul><br></font>
-
<li ><a href="https://2010.igem.org/Team:DTU-Denmark/Characterization_Results"> Characterization Results</a></li>
+
<li ><a href="https://2010.igem.org/Team:DTU-Denmark/AntiTermination_Section">Anti-Terminator Section</a></li>
<ul>
<ul>
<font size="2">
<font size="2">
-
<li><a href="https://2010.igem.org/Team:DTU-Denmark/Characterization_Results_repressors" target="_blank">Repressor Section</a></li>
+
<li><a href="https://2010.igem.org/Team:DTU-Denmark/AntiTermination_Section#Experimental" target="_blank">Experimental Setup</a></li>
-
<li><a href="https://2010.igem.org/Team:DTU-Denmark/Characterization_Results_antiterminators" target="_blank">Anti-Terminators Section</a></li>
+
<li><a href="https://2010.igem.org/Team:DTU-Denmark/AntiTermination_Section#Characterization" target="_blank">Characterization Results</a></li>
</ul><br></font>
</ul><br></font>
</ul>
</ul>
Line 170: Line 170:
<p align="justify">In order to optimize the lab work, we split up the work so that we could have two lab teams working in parallel to design different parts of the switch. We split up the lab work so we had:<br>
<p align="justify">In order to optimize the lab work, we split up the work so that we could have two lab teams working in parallel to design different parts of the switch. We split up the lab work so we had:<br>
<ul>
<ul>
-
<li>Team 1: <b>The Repressor - Anti-Repressor Team</b></li>
+
<li>Team 1: <b>The Repressor - Anti-Repressor Section</b></li>
-
<li>Team 2: <b>The Terminator - Anti-Terminator Team</b></li>
+
<li>Team 2: <b>The Terminator - Anti-Terminator Section</b></li>
</ul>
</ul>
The Repressor (Repressor - Anti-Repressor) Team is responsible for assembling the construct illustrated below in Figure 1:<br>
The Repressor (Repressor - Anti-Repressor) Team is responsible for assembling the construct illustrated below in Figure 1:<br>

Revision as of 09:53, 24 October 2010

Welcome to the DTU iGEM wiki!


Introduction

In order to optimize the lab work, we split up the work so that we could have two lab teams working in parallel to design different parts of the switch. We split up the lab work so we had:

  • Team 1: The Repressor - Anti-Repressor Section
  • Team 2: The Terminator - Anti-Terminator Section
The Repressor (Repressor - Anti-Repressor) Team is responsible for assembling the construct illustrated below in Figure 1:

The Anti-Terminator (Terminator - Anti-Terminator) Team is responsible for assembling the construct illustrated in Figure 2:

Repressor Group

The repressor group will be assembling the constructs step-by-step:

Step 1

The construction of a plasmid containing the divergent promoters is the first step, the effect of this will be the uninhibited expression of GFP as illustrated by the green colonies observed in Figure 4.

Figure 3: The initial plasmid constructed is illustrated. The divergent promoters have been inserted into a plasmid and transformed into the electro-competent E.coli cells.

Figure 3: The success of the plasmid construction and transformation is illustrated by the fluorescent green colonies seen on the LB-agar plates.

Step 2

Figure 4: The construction of a plasmid containing the Repressor protein (GogR or GtgR) expressed from the pRM promoter is shown. The continually expressed repressor protein will then inhibit the pR promoter and no GFP will be expressed.

Step 3

Figure 5: The independent plasmid is constructed is shown. This plasmid contains the gene encoding the anti-repressor is found downstream of the promoter induced by arabinose, pBAD.

Results Simulation

Figure 7: The graphs illustrated are a simulation of the expected results from Construct 2 and Construct 3. The expected results from Construct 2 would be a baseline expression of GFP, as the promoter would continually be repressed. With Construct 3, there would be a baseline expression of GFP until the pBAD is induced. At this point, the anti-repressor is expressed and binds to the repressor preventing its activity. This leads to an increase in the expression of GFP as illustrated by the red curve.

Figure 7: The graphs illustrated are a simulation of the expected results from Construct 2 and Construct 3. The expected results from Construct 2 would be a baseline expression of GFP, as the promoter would continually be repressed. With Construct 3, there would be a baseline expression of GFP until the pBAD is induced. At this point, the anti-repressor is expressed and binds to the repressor preventing its activity. This leads to an increase in the expression of GFP as illustrated by the red curve.