Team:UC Chile/Cyanolux/Project

From 2012.igem.org

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<p>This construct is an integrative plasmid which targets neutral recombination sites (slr0370 and sll0337). We selected this locus because it has been extensively used in the literature ([[#11| 11]]) and it shown to have no deleterious effects on Synechocystis viability. We selected Kanamycin resistance as our selectable marker. [PUT LINK TO CONSTRUCT HERE].</p>
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<p>This construct is an integrative plasmid which targets neutral recombination sites (slr0370 and sll0337). We selected this locus because it has been extensively used in the literature ([[#11| 11]]) and it shown to have no deleterious effects on Synechocystis viability. We selected Kanamycin resistance as our selectable marker.
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Using this backbone we have decided to put LuxAB under the transaldolase promoter.We have choose the transaldolase promoter to express the luciferase part of the operon, as in the literature the promoter is described as having a peak of expression at 2 hours past dusk, which we believe is just the right timing to "turn on the lamp".
Using this backbone we have decided to put LuxAB under the transaldolase promoter.We have choose the transaldolase promoter to express the luciferase part of the operon, as in the literature the promoter is described as having a peak of expression at 2 hours past dusk, which we believe is just the right timing to "turn on the lamp".
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<p>Due to issues mentioned in the results page (PUT LINK HERE) we designed a new plasmid backbone.
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<p>Due to issues mentioned in the [http://2012.igem.org/Team:UC_Chile/Cyanolux/Results results page] we designed a new plasmid backbone.
This is an integration plasmid which makes Synechocystis susceptible to copper concentrations higher than 0.75 uM [[#10|10]] by disrupting the CopS gene. We believe that this strategy serves as a biosafety measure to avoid the possibility of having a leakage of recombinant DNA to the environment. The plasmid uses Spectynomycin as a selectable marker. [PUT LINK TO CONSTRUCT HERE]
This is an integration plasmid which makes Synechocystis susceptible to copper concentrations higher than 0.75 uM [[#10|10]] by disrupting the CopS gene. We believe that this strategy serves as a biosafety measure to avoid the possibility of having a leakage of recombinant DNA to the environment. The plasmid uses Spectynomycin as a selectable marker. [PUT LINK TO CONSTRUCT HERE]
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We plan on expressing LuxCDEG under the control of the promoters Pcaa3 and PsigE (mentioned above). These promoters have peak activities 1 hour before dusk. We believe that we might enhance bioluminescence yield initially by setting the substrate production/regeneration part of the operon prior to the expression of the luciferase.(LINK TO MODELLING?)
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We plan on expressing LuxCDEG under the control of the promoters Pcaa3 and PsigE (mentioned above). These promoters have peak activities 1 hour before dusk. Based on our [http://2012.igem.org/Team:UC_Chile/Cyanolux/Modelling modelling] we believe that we might enhance bioluminescence yield initially by setting the substrate production/regeneration part of the operon prior to the expression of the luciferase.
<h1>References</h1>
<h1>References</h1>

Revision as of 03:56, 27 September 2012

Project: Luxilla - Pontificia Universidad Católica de Chile, iGEM 2012