Team:UC Chile/Cyanolux/Project short

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In 2010 the Cambridge iGEM team Biobricked the LuxBrick, a collection of genes from the Lux operon that incorporate both the Luciferase and the substrate production enzymes without regulation, allowing endogenous bioluminescence on E. coli.
In 2010 the Cambridge iGEM team Biobricked the LuxBrick, a collection of genes from the Lux operon that incorporate both the Luciferase and the substrate production enzymes without regulation, allowing endogenous bioluminescence on E. coli.
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<h3>Chassis</h3>
<h3>Chassis</h3>
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Coupling the endogenous circadian rhythms of this organism to the expression of the Lux genes will enable high-level functionality, through an automatically switching system that turns on bioluminescence only when needed.
Coupling the endogenous circadian rhythms of this organism to the expression of the Lux genes will enable high-level functionality, through an automatically switching system that turns on bioluminescence only when needed.
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<h2>Strategy</h2>
 
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According to literature (CITA!), the limiting step for the bacterial bioluminescent reaction is the substrate (n-decanal) concentration, therefore, to control light emission over time we decided to control it´s abundance in the cells, which in our model is a function of the substrates generation (by Lux C, D, E and G enzymes) and consumption (by the LuxAB luciferase).
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<h2>Strategy</h2>
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The limiting step for the bacterial bioluminescent reaction is the substrate (n-decanal) concentration, therefore, to control light emission over time we decided to control it´s abundance in the cells, which in our model is a function of the substrates generation (by Lux C, D, E and G enzymes) and consumption (by the LuxAB luciferase).
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Our model works as a “black box” in which the input takes the form of a specific hour of the day (i.e the hour on which you want your metabolite to reach maximal concentration) and the output is a couple of promoters from Synechocystis genome.  
Our model works as a “black box” in which the input takes the form of a specific hour of the day (i.e the hour on which you want your metabolite to reach maximal concentration) and the output is a couple of promoters from Synechocystis genome.  
It assumes that the metabolite's production is controlled by enzymes under the control of promoter 1 and it´s degradation by enzymes under promoter 2.
It assumes that the metabolite's production is controlled by enzymes under the control of promoter 1 and it´s degradation by enzymes under promoter 2.
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For more details please check [2012.igem.org/Team:UC_Chile/Cyanolux/Modelling here]
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For more details please check [https://2012.igem.org/Team:UC_Chile/Cyanolux/Modelling here]
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As there weren´t straighforward tools to start working with in the registry (i.e characterized plasmids backbones, protocols, etc) we started from scratch.
As there weren´t straighforward tools to start working with in the registry (i.e characterized plasmids backbones, protocols, etc) we started from scratch.
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We designed two recombination plasmids backbones.  One targets a gene essential for our chassis survival in the environment (link:see biosafety) and the other one a neutral site.
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We designed two recombination plasmids backbones.  One targets a gene essential for our chassis survival in the environment [https://2012.igem.org/Team:UC_Chile/Biosafety#Susceptibility_Construct (see biosafety)] and the other one a neutral site.
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[[File:UC_Chile-IntKstrategy.jpg | 480px | left]]
[[File:UC_Chile-IntKstrategy.jpg | 480px | left]]
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<h2>Implementation</h2>
<h2>Implementation</h2>
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Synthetic biology inspired in nature to make abstractions of it´s principles and mechanisms. We thought this moto could be applied beyond mollecular genetics...
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Synthetic biology inspires in nature making abstractions of its principles and mechanisms. We thought this moto could be applied beyond mollecular genetics...
With the relevance of context in mind, a biomimetic biolamp structure was designed that resembles the organ in which Vibrio fischeri -the bacteria from which the lux genes were biobricked- lives.
With the relevance of context in mind, a biomimetic biolamp structure was designed that resembles the organ in which Vibrio fischeri -the bacteria from which the lux genes were biobricked- lives.
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[https://2012.igem.org/Team:UC_Chile/Cyanolux/Biolamp Full description of the device here]
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[https://2012.igem.org/Team:UC_Chile/Cyanolux/Biolamp Full description of the biolamp device here]
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[https://2012.igem.org/Team:UC_Chile/Cyanolux/Project See more about the whole project]
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<a href="https://2012.igem.org/Team:UC_Chile/Cyanolux/Results_short"><img src="https://static.igem.org/mediawiki/2012/a/ab/UC_Chile-Continue_button.jpg" align="right">
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[https://2012.igem.org/Team:UC_Chile/Cyanolux/Project See more about the project]
 
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Latest revision as of 03:23, 27 October 2012

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