Team:KULeuven/Data

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<li class="off"><a href="https://2011.igem.org/Team:KULeuven/Project">Project</a>
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<li class="off"><a href="https://2011.igem.org/Team:KULeuven/Description">Project</a>
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<li><a href="https://2011.igem.org/Team:KULeuven/Project">Summary</a></li>
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<li><a href="https://2011.igem.org/Team:KULeuven/Description">Description</a></li>
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<li><a href="https://2011.igem.org/Team:KULeuven/Details">Extended</a><li>
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<li><a href="https://2011.igem.org/Team:KULeuven/Modeling">Modeling</a></li>
<li><a href="https://2011.igem.org/Team:KULeuven/Modeling">Modeling</a></li>
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                <li><a href="https://2011.igem.org/Team:KULeuven/Thermodynamics">Thermodynamics</a></li>
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                <li><a href="https://2011.igem.org/Team:KULeuven/Applications">Applications</a></li>
<li><a href="https://2011.igem.org/Team:KULeuven/Biobricks">Biobricks</a></li>
<li><a href="https://2011.igem.org/Team:KULeuven/Biobricks">Biobricks</a></li>
<li><a href="https://2011.igem.org/Team:KULeuven/Notebook">Notebook</a></li>
<li><a href="https://2011.igem.org/Team:KULeuven/Notebook">Notebook</a></li>
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                <li><a href="https://2011.igem.org/Team:KULeuven/Results">Results</a></li>
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<li><a href="https://2011.igem.org/Team:KULeuven/Ethics">Ethics</a></li>
<li><a href="https://2011.igem.org/Team:KULeuven/Ethics">Ethics</a></li>
<li><a href="https://2011.igem.org/Team:KULeuven/Safety">Safety</a></li>
<li><a href="https://2011.igem.org/Team:KULeuven/Safety">Safety</a></li>
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                <li><a href="https://2011.igem.org/Team:KULeuven/Law&Patents">Law&Patents</a></li>
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<img src="http://homes.esat.kuleuven.be/~igemwiki/images/data/figure02.png"><br><br>
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<b>Plasmid 1</b> contains <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584028" target="blank"> <b>BBa_K584028</b> </a> that enables <i>E.D. Frosti</i> to respond immediately to lactose and produce INP.
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<b>Plasmid 1</b> contains the bricks necessary to induce expression of the red color (via the CrtEBI system) and  ice-nucleating protein (INP) in a lactose-dependent manner. For this plasmid, the INP has been cloned (<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584024" target="blank"><b>BBa_K584024</b></a>) and tested for functionality by putting it behind the constitutive (<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584027" target="blank"><b>BBa_K584027</b></a>) and lactose-inducible (<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584028" target="blank"> <b>BBa_K584028</b> </a>) promoter.
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<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584027" target="blank"><b>BBa_K584027</b></a> is a precursor of <b>BBa_K584028</b>, with a constitutive promoter.
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<b>Plasmid 2</b> contains the bricks that induce the expression of the black color (via the melA system) and  antifreeze protein (AFP) in a arabinose-dependent manner. For this plasmid, the AFP has been cloned (<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584020" target="blank"> <b>BBa_K584020</b></a>) and tested for functionality by putting it behind the constitutive promoter (<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584026" target="blank"><b>BBa_K584026</b></a>).<br><br>
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<b>Plasmid 2</b> contains <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584020" target="blank"> <b>BBa_K584020</b> </a> for AFP production. We were able to put AFP behind a constitutive promoter (<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584026" target="blank"><b>BBa_K584026</b></a>) but due to lack of time we could not finish the AFP testing. <br>
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<b>Plasmid 3</b> contains <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584004" target="blank"><b>BBa_K584004</b></a>. This is a GFP-construct to test the activity of the HybB promoter. Due to lack of time we could not test this biobrick. We wanted to engineer a biobrick in which the ribokey is controlled by the HybB promoter.
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<b>Plasmid 3</b> contains the necessary bricks for our cell death mechanism. Briefly, lactose or arabinose will generate a ribolock-ceaB construct (<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584015" target="blank" ><b> BBa_K584015</b></a> and <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584016" target="blank"> <b>BBa_K584016</b></a>).The Ribokey can be activated by a cold shock(<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584014" target="blank"> <b>BBa_K584014</b></a>). For this plasmid we cloned the lactose- and arabinose-inducible ceaB construct (<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584016" target="blank"> <b>BBa_K584016</b></a> and <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584015" target="blank"> <b>BBa_K584015</b></a> resp.) and characterized the cold shock promoter using the Biobrick <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584004" target="blank"><b>BBa_K584004</b></a>.
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<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584027" target="blank"> <b>BBa_K584027</b> </a> was inserted in plasmid 1 to characterize INP.
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<img src="http://homes.esat.kuleuven.be/~igemwiki/images/data/figure03.png"><br><br>
<img src="http://homes.esat.kuleuven.be/~igemwiki/images/data/figure03.png"><br><br>
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All our obtained results can also be found at our <a href="https://2011.igem.org/Team:KULeuven/Results">results</a> section.
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<br><br>
<h2>Data for our favorite new parts</h2>
<h2>Data for our favorite new parts</h2>
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<li> 1. <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584020" target="blank"> <b>BBa_K584020</b>  </a> - <b> AFP extracellular protein generator </b> (K.U.Leuven, iGEM 2011): this biobrick generates AFP that will be expressed extracellularly. We tested this biobrick by putting it behind a constitutive promoter (which yielded <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584026" target="blank"><b>BBa_K584026</b></a> ).
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<li> 1. <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584024" target="blank"> <b>BBa_K584024</b>  </a> - <b> INP extracellular generator </b> (K.U.Leuven, iGEM 2011): this biobrick generates INP that is expressed extracellularly. We tested this biobrick by putting it behind a constitutive promoter (which yielded <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584027" target="blank"><b>BBa_K584027</b></a>) and the lactose-inducible promoter (yielding  <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584028" target="blank"><b>BBa_K584028</b></a>).
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<li> 2. <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584024" target="blank"> <b>BBa_K584024</b> </a> - <b> INP generator </b> (K.U.Leuven, iGEM 2011): this biobrick generates INP that is expressed extracellularly. We tested this biobrick by putting it behind a constitutive promoter (which yielded <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584027" target="blank"> <b>BBa_K584027</b></a>).
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<li> 2. <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584020" target="blank"> <b>BBa_K584020</b> </a> - <b> AFP extracellular protein generator </b> (K.U.Leuven, iGEM 2011): this biobrick generates AFP that will be expressed extracellularly. We tested this biobrick by putting it behind a constitutive promoter (which yielded <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584026" target="blank"> <b>BBa_K584026</b></a>).
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<h2> Data for pre-existing parts</h2>
<h2> Data for pre-existing parts</h2>
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<li> 1.  <a href="http://partsregistry.org/Part:BBa_I13453:Experience#User_Reviews" target="blank"> <b>BBa_I13453</b> </a> - <b> pBAD promoter</b> (Endy Lab, iGEM 2005): fused a GFP reporter to this promoter (which created biobrick: <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584000" target="blank"> <b>BBa_K584000</b></a>). The results can be found at the experience page of that promoter.
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<li> 1.  <a href="http://partsregistry.org/Part:BBa_I13453:Experience#User_Reviews" target="blank"> <b>BBa_I13453</b> </a> - <b> pBAD promoter</b> (Endy Lab, iGEM 2005): fused a GFP reporter to this promoter (which created biobrick: <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584000" target="blank"> <b>BBa_K584000</b></a>). The results can be found at the experience page of the promoter.
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<li> 2. <a href="http://partsregistry.org/Part:BBa_J23119:Experience" target="blank"><b>BBa_J23119</b> </a> - <b> constitutive promoter</b> (Berkeley, iGEM 2006): fused a GFP reporter to this promoter (which created biobrick: <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584001" target="blank"> BBa_K584001</a>). The results can be found at the experience page of that promoter.
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<li> 2. <a href="http://partsregistry.org/Part:BBa_J23119:Experience" target="blank"><b>BBa_J23119</b> </a> - <b> constitutive promoter</b> (Berkeley, iGEM 2006): fused a GFP reporter to this promoter (which created biobrick: <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584001" target="blank"><b>BBa_K584001</b> </a>). The results can be found at the experience page of the promoter.
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<li> 3. <a href="http://partsregistry.org/Part:BBa_K091100:Experience" target="blank"><b>BBa_K091100</b> </a> - <b> Lac-Lux hybrid promoter</b> (Davidson-Missouri_Western, iGEM 2008): fused a GFP to this promoter (which created biobrick <a href="http://partsregistry.org/Part:BBa_K584002" target="blank"><b>BBa_K584002</b> </a>). The results can be found at the experience page of the promoter.
<h2>We've also characterized the following parts</h2>
<h2>We've also characterized the following parts</h2>
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<li> 1. <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584004" target="blank"> <b>BBa_K584004</b> </a> - <b> HybB promoter + GFP generator</b> (K.U.Leuven, iGEM 2011): we created this biobrick but due to lack of time we could not test it.
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<li> 1. <a href="http://partsregistry.org/Part:BBa_J45503:Experience" target="blank"> <b>BBa_J45503</b> </a> - <b> HybB promoter</b> (MIT, iGEM 2006): fused a GFP to this promoter (which yielded biobrick <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584004" target="blank"> <b>BBa_K584004</b> </a>). The results can be found at the experience page of the promoter.<br><br>
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<li> 2. <a href="http://partsregistry.org/Part:BBa_K091107:Experience" target="blank"> <b>BBa_K091107</b> </a> - <b> pLux/cI Hybrid Promoter</b> (Davidson-Missouri_Western, iGEM 2008): we wanted to use this promoter at the inception of our project, however on a 1% agarose gel we saw fragments of a different height than reported by previous iGEM competitions.<br><br>
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<li> 2. <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584027" target="blank"> <b>BBa_K584027</b></a>, <b>Constitutive promoter + INP </b>(K.U.Leuven, iGEM 2011): we characterized this promoter by coupling a GFP behind it. The characterization can be found on the registry page of the original biobrick: <a href="http://partsregistry.org/Part:BBa_J23119:Experience" target="blank"><b> BBa_J23119 </b></a>.
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<li> 3. <a href="http://partsregistry.org/Part:BBa_K091104:Experience" target="blank"> <b>BBa_K091104</b> </a> - <b> pLac/Mnt Hybrid Promoter</b> (Davidson-Missouri_Western, iGEM 2008): we wanted to use this promoter at the inception of our project, however on a 1% agarose gel we saw fragments at a different height than reported by previous iGEM competitions.
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<li> 3. <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K584026" target="blank"><b>BBa_K584026</b></a>, <b>Constitutive promoter + AFP </b> (K.U.Leuven, iGEM 2011): For this biobrick, we used the same constitutive promoter (<a href="http://partsregistry.org/Part:BBa_J23119:Experience" target="blank"><b> BBa_J23119 </b></a>), the characterization of this biobrick was the same.
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Latest revision as of 07:45, 28 October 2011

KULeuven iGEM 2011

close

Data page

How does E.D. Frosti work?



The first illustration is a global overview of our project in which you can see how the plasmids (and the parts) function in our system. The next illustration shows the plasmids in more detail, with the favorite new parts and the characterized pre-existing parts.


Plasmid 1 contains the bricks necessary to induce expression of the red color (via the CrtEBI system) and ice-nucleating protein (INP) in a lactose-dependent manner. For this plasmid, the INP has been cloned (BBa_K584024) and tested for functionality by putting it behind the constitutive (BBa_K584027) and lactose-inducible ( BBa_K584028 ) promoter.

Plasmid 2 contains the bricks that induce the expression of the black color (via the melA system) and antifreeze protein (AFP) in a arabinose-dependent manner. For this plasmid, the AFP has been cloned ( BBa_K584020) and tested for functionality by putting it behind the constitutive promoter (BBa_K584026).

Plasmid 3 contains the necessary bricks for our cell death mechanism. Briefly, lactose or arabinose will generate a ribolock-ceaB construct ( BBa_K584015 and BBa_K584016).The Ribokey can be activated by a cold shock( BBa_K584014). For this plasmid we cloned the lactose- and arabinose-inducible ceaB construct ( BBa_K584016 and BBa_K584015 resp.) and characterized the cold shock promoter using the Biobrick BBa_K584004.


All our obtained results can also be found at our results section.

Data for our favorite new parts

  • 1. BBa_K584024 - INP extracellular generator (K.U.Leuven, iGEM 2011): this biobrick generates INP that is expressed extracellularly. We tested this biobrick by putting it behind a constitutive promoter (which yielded BBa_K584027) and the lactose-inducible promoter (yielding BBa_K584028).

  • 2. BBa_K584020 - AFP extracellular protein generator (K.U.Leuven, iGEM 2011): this biobrick generates AFP that will be expressed extracellularly. We tested this biobrick by putting it behind a constitutive promoter (which yielded BBa_K584026).

    Data for pre-existing parts

  • 1. BBa_I13453 - pBAD promoter (Endy Lab, iGEM 2005): fused a GFP reporter to this promoter (which created biobrick: BBa_K584000). The results can be found at the experience page of the promoter.

  • 2. BBa_J23119 - constitutive promoter (Berkeley, iGEM 2006): fused a GFP reporter to this promoter (which created biobrick: BBa_K584001 ). The results can be found at the experience page of the promoter.

  • 3. BBa_K091100 - Lac-Lux hybrid promoter (Davidson-Missouri_Western, iGEM 2008): fused a GFP to this promoter (which created biobrick BBa_K584002 ). The results can be found at the experience page of the promoter.

    We've also characterized the following parts

  • 1. BBa_J45503 - HybB promoter (MIT, iGEM 2006): fused a GFP to this promoter (which yielded biobrick BBa_K584004 ). The results can be found at the experience page of the promoter.

  • 2. BBa_K091107 - pLux/cI Hybrid Promoter (Davidson-Missouri_Western, iGEM 2008): we wanted to use this promoter at the inception of our project, however on a 1% agarose gel we saw fragments of a different height than reported by previous iGEM competitions.

  • 3. BBa_K091104 - pLac/Mnt Hybrid Promoter (Davidson-Missouri_Western, iGEM 2008): we wanted to use this promoter at the inception of our project, however on a 1% agarose gel we saw fragments at a different height than reported by previous iGEM competitions.