Team:Colombia/Notebook/Characterization

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==Characterization==
==Characterization==
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Our objective was to analyze the of our system for chitin detection and activation of the chininolytic metabolism of ''Vibrio fisherii'' ES144, we proposed two different strategies:
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First, we needed to characterize the expression of the two constitutive promoters of the two-component system (Sensor Promoter and CBP Promoter).
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We made a construct where the CBP promoter leads the expression of the mCherry reporter. The characterization was made using fluorometry and using as control the constitutive promoter of ''E. coli'' (BBa_J23119).
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The characterization of the chitin sensor on ''E. coli'' will be made on the future using the same strategy.
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Aditionally, we needed to made the characterization of the inducible promoter of the two component system (ChiP) as a response of the extracellular chitin concentration.
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In order to do that we realized a construct where ChiP was Upstream from the mCherry reporter. Without a termination codon, mCherry expression is controlled by the ChiP promoter.
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Este constructo se introducirá en una bacteria que contenta también plásmidos que codifican para el Sensor y para CBP, y el nivel de fluorescencia será una medida de la activación del promotor inducible a diferentes concentraciones extracelulares de quitina.
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Para esta caracterización los vectores ya fueron construidos y la transformación se realizara prontamente, los resultados se los mostramos en el Jamboree;)
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[[File:CharacColombia.png]]
[[File:CharacColombia.png]]

Revision as of 03:36, 29 October 2011

Template:Https://2011.igem.org/User:Tabima

Characterization

Our objective was to analyze the of our system for chitin detection and activation of the chininolytic metabolism of Vibrio fisherii ES144, we proposed two different strategies:

First, we needed to characterize the expression of the two constitutive promoters of the two-component system (Sensor Promoter and CBP Promoter). We made a construct where the CBP promoter leads the expression of the mCherry reporter. The characterization was made using fluorometry and using as control the constitutive promoter of E. coli (BBa_J23119).

The characterization of the chitin sensor on E. coli will be made on the future using the same strategy.

Aditionally, we needed to made the characterization of the inducible promoter of the two component system (ChiP) as a response of the extracellular chitin concentration. In order to do that we realized a construct where ChiP was Upstream from the mCherry reporter. Without a termination codon, mCherry expression is controlled by the ChiP promoter.

Este constructo se introducirá en una bacteria que contenta también plásmidos que codifican para el Sensor y para CBP, y el nivel de fluorescencia será una medida de la activación del promotor inducible a diferentes concentraciones extracelulares de quitina. Para esta caracterización los vectores ya fueron construidos y la transformación se realizara prontamente, los resultados se los mostramos en el Jamboree;)

CharacColombia.png