"
Team:ETH Zurich/pre proc
From 2013.igem.org
| Line 8: | Line 8: | ||
<b>Natural Lux signalling system</b> | <b>Natural Lux signalling system</b> | ||
<br> | <br> | ||
| - | <p align="justify">As our system is based on a sender-receiver module, we use the lux signalling system in our bio-game. In <i>Vibriofischeri</i>, the <i>lux</i> gene encodes for the enzyme that catalyses the production of the signalling molecule N-acyl-homoserine lactone, <b>AHL</b>. The AHL molecule is able to <b>diffuse</b> in and out of cells. | + | <p align="justify">As our system is based on a sender-receiver module, we use the lux signalling system in our bio-game. In <i>Vibriofischeri</i>, the <i>lux</i> gene encodes for the enzyme that catalyses the production of the signalling molecule N-acyl-homoserine lactone, <b>AHL</b>. The AHL molecule is able to <b>diffuse</b> in and out of cells. Meanwhile, the LuxR protein is produced which is driven by P<sub>luxL</sub>. When unbound, this protein is in the inactive state. The binding of AHL to LuxR protein converts inactive LuxR to an <b>active LuxR-AHL complex</b>. |
| - | + | The complex binds to the P<sub>LuxR</sub> promoter and drives the gene expression forward .[http://www.fasebj.org/content/7/11/1016. 1]</p> | |
<br> | <br> | ||
Revision as of 21:25, 27 October 2013
Contents |
LuxR signaling system
Natural Lux signalling system
As our system is based on a sender-receiver module, we use the lux signalling system in our bio-game. In Vibriofischeri, the lux gene encodes for the enzyme that catalyses the production of the signalling molecule N-acyl-homoserine lactone, AHL. The AHL molecule is able to diffuse in and out of cells. Meanwhile, the LuxR protein is produced which is driven by PluxL. When unbound, this protein is in the inactive state. The binding of AHL to LuxR protein converts inactive LuxR to an active LuxR-AHL complex. The complex binds to the PLuxR promoter and drives the gene expression forward .[http://www.fasebj.org/content/7/11/1016. 1]
Exploiting the lux signalling system in Colisweeper
In the bio-game, the mines are the sender cells and the non-mines are the receiver cells. The mines constitutively express LuxI under [http://parts.igem.org/Part:BBa_J23110 promoter (Part BBa_J23110)] and hence AHL. This molecule then diffuses through the agar and can be processed by the non-mines. The non-mines produce luxR under [http://parts.igem.org/Part:BBa_J09855 promoter (Part BBa_J09855)] and so the incoming AHL will bind to the luxR protein to form an active complex AHL-luxR. The non-mines also contain PLuxR promoters that are tuned to different AHL sensitivities. The AHL-luxR complex will bind to the promoters and drive the expression of the reporter hydrolases. Depending on the amount of AHL that is processed, different expression of output colors give the player the cue for the next move in the bio-game.
Diffusion of Signalling molecule AHL from mine to non-mine
In order to study the communication between the mines and non-mines via the signalling molecule AHL, diffusion tests were performed. It was vital to know the exact diffusion pattern of AHL from the mine to have an idea of the placement of the colonies on the agar-mine field. By studying the diffusion time and distance of AHL around the sender, we were able to confirm the honey-comb pattern as the design of the mine grid. With this design, colonies are placed on the edges of each hexagon in the honey-comb pattern. See results of these experiments here.
