Team:Tec-Monterrey/projectmodeling

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        <p><a href="https://2011.igem.org/Team:Tec-Monterrey/projectoverview">overview</a></p>
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                <p><a href="https://2011.igem.org/Team:Tec-Monterrey/projectoverview">overview</a></p>
             <p><a href="https://2011.igem.org/Team:Tec-Monterrey/projectparts">parts</a></p>
             <p><a href="https://2011.igem.org/Team:Tec-Monterrey/projectparts">parts</a></p>
             <p><a href="https://2011.igem.org/Team:Tec-Monterrey/projectmodeling">genetic frame</a></p>
             <p><a href="https://2011.igem.org/Team:Tec-Monterrey/projectmodeling">genetic frame</a></p>
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             <p><a href="https://2011.igem.org/Team:Tec-Monterrey/safetypage">safety</a></p>
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             <p><a href="https://2011.igem.org/Team:Tec-Monterrey/projectresults/methods">methods</a></p>
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            <p><a href="https://2011.igem.org/Team:Tec-Monterrey/teamha">human practice</a></p>
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            <p><a href="https://2011.igem.org/Team:Tec-Monterrey/projectnotebook">notebook</a></p>
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             <p><a href="https://2011.igem.org/Team:Tec-Monterrey/projectresults">results</a></p>
             <p><a href="https://2011.igem.org/Team:Tec-Monterrey/projectresults">results</a></p>
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            <p><a href="https://2011.igem.org/Team:Tec-Monterrey/teamha">human approach</a></p>
             <p><a href="https://2011.igem.org/Team:Tec-Monterrey/projectprotocols">protocols</a><p>
             <p><a href="https://2011.igem.org/Team:Tec-Monterrey/projectprotocols">protocols</a><p>
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            <p><a href="https://2011.igem.org/Team:Tec-Monterrey/safetypage">safety</a></p>
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            <p><a href="https://2011.igem.org/Team:Tec-Monterrey/projectnotebook">notebook</a></p>
             <p><a href="https://2011.igem.org/Team:Tec-Monterrey/sampledata">sample data</a></p>
             <p><a href="https://2011.igem.org/Team:Tec-Monterrey/sampledata">sample data</a></p>
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  The first DNA construction our team used, was called “Construct C” this assembly was made in order to obtain data from our AraC-Pbad promoter. Our team worked with parts from British Columbia 09 team. Part BBa_K206000, pBAD,  is an E.coli promoter that is induced by L-arabinose. In the absence of arabinose, the repressor protein AraC, BBa_I13458, binds to the AraI1 operator site of pBAD and the upstream operator site AraO2, blocking transcription [1]. In the presence of arabinose, AraC binds to it and changes its conformation such that it interacts with the AraI1 and AraI2 operator sites, permitting transcription.
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  The first DNA construction our team used, was called “Construct GFP” this assembly was made in order to obtain data from our araC-P<sub>BAD</sub> promoter. Our team worked with parts from British Columbia 09 team. Part <a href="http://partsregistry.org/Part:BBa_K206000">BBa_K206000</a>, P<sub>BAD</sub>,  is an <i>Escherichia coli</i> promoter that is induced by L-arabinose. In the absence of arabinose, the repressor protein AraC, <a href="http://partsregistry.org/Part:BBa_I13458">BBa_I13458</a>, binds to the AraI1 operator site of P<sub>BAD</sub> and the upstream operator site AraO2, blocking transcription [1]. In the presence of arabinose, AraC binds to it and changes its conformation such that it interacts with the AraI1 and AraI2 operator sites, permitting transcription.
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K206000 is a variant pBAD promoter with a modified AraI1 site that has been shown both to be responsive to lower concentrations of arabinose and to exhibit a higher maximum expression than the wild type (BBa_I13453), as measured by coupling to a fluorescent reporter by iGEM09 British Columbia Team.
 
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We aslo used as a RBS the Kozak sequence used normally on most constructs, part BBa_B0034, followed by a Green Fluorecent Protein (BBa_b0040), supposed to act as a reporter in order to meassure effectivness of the arabinose promoter. This construct was meassured in e. coli’s strain BW27783, which is unable to metabolize arabinose, (our inductor).
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<a href="http://partsregistry.org/Part:BBa_K206000">BBa_K206000</a> is a variant pBAD promoter with a modified AraI1 site that has been shown both to be responsive to lower concentrations of arabinose and to exhibit a higher maximum expression than the wild type (<a href="http://partsregistry.org/Part:BBa_I13453">BBa_I13453</a>), as measured by coupling to a fluorescent reporter by iGEM09 British Columbia Team.
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Our results meassured by our team member Fausto Ortiz, were the next ones:
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We aslo used as a RBS the Kozak sequence used normally on most constructs, part <a href="http://partsregistry.org/Part:BBa_B0034">BBa_B0034</a>, followed by a Green Fluorecent Protein (<a href="http://partsregistry.org/Part:BBa_B0040">BBa_B0040</a>), supposed to act as a reporter in order to meassure effectivness of the arabinose promoter. This construct was meassured in <i>E. coli</i> strain BW27783, which is unable to metabolize L-arabinose (our inductor).
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Schlief, R. (2000). Regulation of the L-arabinose operon of Escherichia coli. Trends in Genetics. 16(12):559-565.
 
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Schlief, R. (2000). Regulation of the L-arabinose operon of <i>Escherichia coli</i>. Trends in Genetics. 16(12):559-565.
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Latest revision as of 20:50, 20 October 2011

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iGEM

 

 


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The first DNA construction our team used, was called “Construct GFP” this assembly was made in order to obtain data from our araC-PBAD promoter. Our team worked with parts from British Columbia 09 team. Part BBa_K206000, PBAD, is an Escherichia coli promoter that is induced by L-arabinose. In the absence of arabinose, the repressor protein AraC, BBa_I13458, binds to the AraI1 operator site of PBAD and the upstream operator site AraO2, blocking transcription [1]. In the presence of arabinose, AraC binds to it and changes its conformation such that it interacts with the AraI1 and AraI2 operator sites, permitting transcription.

BBa_K206000 is a variant pBAD promoter with a modified AraI1 site that has been shown both to be responsive to lower concentrations of arabinose and to exhibit a higher maximum expression than the wild type (BBa_I13453), as measured by coupling to a fluorescent reporter by iGEM09 British Columbia Team.

We aslo used as a RBS the Kozak sequence used normally on most constructs, part BBa_B0034, followed by a Green Fluorecent Protein (BBa_B0040), supposed to act as a reporter in order to meassure effectivness of the arabinose promoter. This construct was meassured in E. coli strain BW27783, which is unable to metabolize L-arabinose (our inductor).



Schlief, R. (2000). Regulation of the L-arabinose operon of Escherichia coli. Trends in Genetics. 16(12):559-565.





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