Team:ETH Zurich/achievements

From 2013.igem.org

(Difference between revisions)
Line 4: Line 4:
<h1> We achieved </h1>
<h1> We achieved </h1>
<b>Pre-Processing:</b><br><br>
<b>Pre-Processing:</b><br><br>
-
- Characterize the AHL diffusion on agar plates and define, with the predition from the model and the experimental     results, the distance between colonies, the strengh of the LuxI promoter and the incubation time for the diffusion.<br>
+
- Characterize the AHL diffusion on agar plates and define, with the predition from the model and the experimental results, the distance between colonies, the strengh of the promoter controlling LuxI production and the incubation time for the diffusion.<br>
- To astablish several AHL gradients from different mines on one plate, intersections of gradients result in higher AHL levels for the detection of 1, 2 or 3 mines.<br>
- To astablish several AHL gradients from different mines on one plate, intersections of gradients result in higher AHL levels for the detection of 1, 2 or 3 mines.<br>
- Make a spatio-temporal model of AHL difussion<br><br>
- Make a spatio-temporal model of AHL difussion<br><br>
Line 15: Line 15:
- We measured the dose-response of those promoters in liquid culture as well as on agar plates and determined the EC<sub>50</sub> going from 0.02nM to 6482nM in liquid culture and from 4.45nM to 12'555nM on agar plates.<br>
- We measured the dose-response of those promoters in liquid culture as well as on agar plates and determined the EC<sub>50</sub> going from 0.02nM to 6482nM in liquid culture and from 4.45nM to 12'555nM on agar plates.<br>
- By the prediction of the model we chooose xy to be the right one for our set-up.<br><br>
- By the prediction of the model we chooose xy to be the right one for our set-up.<br><br>
-
- A proof-of-principle using a GFP reporter.<br><br>
+
- a proof-of-principle using a GFP reporter.<br><br>
   
   
Line 21: Line 21:
<b>Optimization:</b>
<b>Optimization:</b>
<br><br>
<br><br>
-
- Identification of the leakyness source.<br>
+
- the identification of the leakyness source.<br>
-
- Optimize the circuit to reduce the leakiness by introduction of a negative feedback loop.<br>
+
- to model different solution to reduce the leakiness
 +
- to optimize the circuit to reduce the leakiness by introduction of a negative feedback loop.<br>
   
   
<b>Output:</b><br><br>
<b>Output:</b><br><br>
-
- Implement several different reporters (hydrolases) in one construct.<br><br>
+
- to implement several different reporters (hydrolases and fluorencent proteins) in one construct.<br><br>
-
- Different colormetric response for all hydrolases.<br>
+
- to show different colormetric (yellow, salmon, violett, green, blue) response for 5 hydrolases.<br>
-
- Characterize the hydrolases by Michaelis-Menten kinetics.<br>
+
- to show the orthogonality of all 5 hydrolases.<br>
 +
- to characterize the hydrolases by Michaelis-Menten kinetics.<br>
<br><br>
<br><br>
<i> A preliminary game with the hydrolases.</i><br><br>
<i> A preliminary game with the hydrolases.</i><br><br>

Revision as of 11:28, 28 October 2013

Header2.png
80px-Eth igem logo.png

We achieved

Pre-Processing:

- Characterize the AHL diffusion on agar plates and define, with the predition from the model and the experimental results, the distance between colonies, the strengh of the promoter controlling LuxI production and the incubation time for the diffusion.
- To astablish several AHL gradients from different mines on one plate, intersections of gradients result in higher AHL levels for the detection of 1, 2 or 3 mines.
- Make a spatio-temporal model of AHL difussion


Processing:

- To isolate a first PLuxR by site directed mutagenisis. - Find a analytical solution for the EC50 of the promoter we need to find.
- To make a promoter libary by partial sensitivity recovery based on the first PLuxR variant and finally isolate 8 promoters with different EC50.
- We measured the dose-response of those promoters in liquid culture as well as on agar plates and determined the EC50 going from 0.02nM to 6482nM in liquid culture and from 4.45nM to 12'555nM on agar plates.
- By the prediction of the model we chooose xy to be the right one for our set-up.

- a proof-of-principle using a GFP reporter.


Optimization:

- the identification of the leakyness source.
- to model different solution to reduce the leakiness - to optimize the circuit to reduce the leakiness by introduction of a negative feedback loop.


Output:

- to implement several different reporters (hydrolases and fluorencent proteins) in one construct.

- to show different colormetric (yellow, salmon, violett, green, blue) response for 5 hydrolases.
- to show the orthogonality of all 5 hydrolases.
- to characterize the hydrolases by Michaelis-Menten kinetics.


A preliminary game with the hydrolases.

Establish a spatio-temporal model of the proof-of-principle and the final game.