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Team:Grenoble/Projet/Design
From 2011.igem.org
Design and principle
Toggle switch explanation
In a few words
Our project is based on a toggle switch system. So we have to explain how it works. Basically it corresponds to an irreversible choice between two pathways depending on the environment. In reality, the reversibility is possible but very difficult to obtain because of the strength of the interactions and the stability of the system (defined by hysteresis * insérer un lien ici*).
Genetically, what is happening ?
Contrary to us, bacteria can’t think or make voluntary choices. So how can they choose a pathway? Through our project, we are creating a genetic system which helps bacteria to take such a decision. For that, we exploit chemical components of the environment which influence the bacterial behaviour. By taking into account two proteins repressing each other’s expression (LacI and XR), we create the fundaments of a toggle system that can switch when using two inducers (IPTG and X) in the medium.
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Suppose that initially the genes in the Toggle Switch are both expressed. |
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If the effect of LacI is dominant through the presence of a high level of IPTG, the bacteria will synthesise more XR and switch off the expression of lacI |
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The bacteria toggle to one side and totally block the expression of lacI. |
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We need to use very high level of the second inducer X to try to switch the toggle back because the active pathway creates an excess of XR, inhibiting lacI expression. |
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The amount of X would have to be so high as to relieve the inhibition of lacI expression and consequently turn off the other pathway |
The toggle switch offers the bacteria the capacity to choose between two pathways in function of their environment. Thanks to that our E.coli can activate one of two Quorum Sensing (*insérer un lien ici*) ways.
Quorum Sensing
- Ultra violet lamp:
- There is a risk for the eyes and the skin, but the UV lamp is only used to take a picture of our gel after electrophoresis, so we are never directly exposed because there is a protective cover and we wear a mask that shields from UV.
- Centrifuge:
- The centrifuges have to be perfectly balanced. All the centrifuges in our lab have detectors that warn the operator in case of imbalance.
- Autoclave:
- The operation of the autoclave require a specific training and has only be performed by trained people.
