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Team:Brasil-SP/Project/DiagnosisModule
From 2014.igem.org
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<h3 align="center">Diagnosis Module</h3> | <h3 align="center">Diagnosis Module</h3> | ||
| - | <p><div align="justify"> If the concentration of Cystatin C is normal, the activity of Cathepsin S is not sufficiently inhibited. Thus, the linker will be cleaved, releasing AIP. The AIP activates ComD, which, in turn, phosphorylates ComE. ComE activates the LasR production, so that it achieves optimal concentration to activate the promoter, even if several LasR proteins interact with qteE. Therefore, the transcription of the reporter gene, GFP, is activated. If the concentration of Cystatin C is above normal, indicating a possible kidney disease, the activity of Cathepsin S is inhibited. In this way, there will not be cleavage of the linker nor the release of AIP. Thus the chain of events is not triggered, so there is no production of the reporter gene.</div></p> | + | <p><div align="justify"> It is crucial for our biodetector to have the ability to discriminate between normal and abnormal Cys C, not just responding to the presence or absence of the same. The interaction of the proteins LasR and QteE is the one responsible for giving our circuit this feature. The QteE destabilize the LasR protein, removing its activity as an inducer, or in other words creating an expression barrier. In our system, the LasR production will be under control of the ComE responsive promoter, while the QteE will be under control of the lactose promoter (IPTG inducible). Having said that, we have to adjust the IPTG concentration so that only the LasR concentration originated from increased Cys C levels. Once we have figured out the appropriate IPTG amount our bacteria will be capable of generating different outputs for normal and increased Cys C levels. |
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| + | <!--If the concentration of Cystatin C is normal, the activity of Cathepsin S is not sufficiently inhibited. Thus, the linker will be cleaved, releasing AIP. The AIP activates ComD, which, in turn, phosphorylates ComE. ComE activates the LasR production, so that it achieves optimal concentration to activate the promoter, even if several LasR proteins interact with qteE. Therefore, the transcription of the reporter gene, GFP, is activated. If the concentration of Cystatin C is above normal, indicating a possible kidney disease, the activity of Cathepsin S is inhibited. In this way, there will not be cleavage of the linker nor the release of AIP. Thus the chain of events is not triggered, so there is no production of the reporter gene.</div></p>--> | ||
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Revision as of 21:46, 11 October 2014
Diagnosis Module
It is crucial for our biodetector to have the ability to discriminate between normal and abnormal Cys C, not just responding to the presence or absence of the same. The interaction of the proteins LasR and QteE is the one responsible for giving our circuit this feature. The QteE destabilize the LasR protein, removing its activity as an inducer, or in other words creating an expression barrier. In our system, the LasR production will be under control of the ComE responsive promoter, while the QteE will be under control of the lactose promoter (IPTG inducible). Having said that, we have to adjust the IPTG concentration so that only the LasR concentration originated from increased Cys C levels. Once we have figured out the appropriate IPTG amount our bacteria will be capable of generating different outputs for normal and increased Cys C levels.
