"
Team:Tec-Monterrey/projectmodeling
From 2011.igem.org
| Line 612: | Line 612: | ||
<p class="textojustif"> | <p class="textojustif"> | ||
| - | 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. | + | 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. |
<br> | <br> | ||
<br> | <br> | ||
| - | <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. | + | <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. |
<br> | <br> | ||
<br> | <br> | ||
| - | 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). | + | 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). |
<br> | <br> | ||
Revision as of 20:48, 20 October 2011
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.
