Team:KU Leuven/Parts

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<div class="span4 icon white">
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    <a href="http://2013.igem.org/Team:KU_Leuven/Project/Aphid_Background">
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    <i class="livicon activeicon" data-name="bug" data-color="white"></i></div>
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  <div class="span7 icon-text">
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    <h3>Aphid Background</h3>
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    </a>
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    <p>Crashcourse in aphid biology</p>
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  </div>
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    <a href="http://2013.igem.org/Team:KU_Leuven/Project/HoneydewSystem">
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    <i class="livicon activeicon" data-name="drop" data-color="white"></i></div>
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  <div class="span7 icon-text">
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    <h3>Honeydew System</h3> </a>
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    <p>Our BanAphids react to honeydew</p>
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  </div>
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  </div>
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    <a href="http://2013.igem.org/Team:KU_Leuven/Project/StickerSystem">
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    <i class="livicon activeicon" data-name="refresh" data-color="white"></i>
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    </div>
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  <div class="span7 icon-text">
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    <h3>Sticker System</h3>
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    </a>
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    <p>BanAphid population oscillates!</p>
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  </div>
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  </div>
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    <a href="http://2013.igem.org/Team:KU_Leuven/Project/E.coligy">
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    <i class="livicon activeicon" data-name="leaf" data-color="white"></i>
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  <div class="span7 icon-text">
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    <h3><i>E. coligy</i></h3>
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    </a>
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    <p>Validating the BanAphids in vivo</p>
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  </div>
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  </div>
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  <a href="http://2013.igem.org/Team:KU_Leuven/Parts">
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    <i class="livicon activeicon" data-name="gears" data-color="white"></i></div>
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  <div class="span7 icon-text">
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    <h3>Parts</h3> </a>
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    <p>You are here!</p>
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  </div>
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    <a href="http://2013.igem.org/Team:KU_Leuven/Project/DataPage">
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    <i class="livicon activeicon" data-name="info" data-color="white"></i>
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    </div>
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  <div class="span7 icon-text">
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    <h3>Data Page</h3> </a>
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    <p>All our achievements on one small page!</p>
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  <p align = "justify">Here you will find the BioBricks we made this summer in the wetlab. Our three favorite parts are marked with a heart:<br/>
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<h3>Favourite parts</h3>
<ol>
<ol>
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<li> BBa_K1060002 was a milestone in our project work. We succeeded to clone the beta farnesene synthase of <i>Artemisia annua</i> in <i>E. coli</i>. To apply to the iGEM standards, we removed an <i>EcoRI</i> site in the gene (AY835398.1).</li><br />
 
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<li> BBa_K1060009 is a construct that constitutively expresses beta farnesene synthase. This synthase will convert farnesyl pyrophosphate into the aphid alarm hormone, (E)-β-farnesene.</li><br />
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<li> <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1060002">BBa_K1060002</a> contains an open reading frame that codes for a sesquiterpene synthase (GenBank Accession No. AY835398), (E)-b-farnesene synthase, that has been isolated from <i>Artemisia annua</i> (Picaud <i>et al.</i>, 2005). The enzyme converts farnesyl diphosphate into E-β-farnesene. E-β-farnesene is an alarm hormone produced by aphids. It is also an important semiochemical in aphid localization. It is released in the cornicle secretions of many aphid species (Franscis <i>et al.</i>, 2005) to alert surrounding aphids of the presence of natural enemies (Kunert <i>et al.</i>, 2005). Detection of these short range chemical cues leads not to the aphid directly, but only indicates the presence, improving prey detection of the predators and parasitoids.<br/><b>Group favourite</b>: it was a milestone in our project work. We succeeded to clone the beta farnesene synthase of <i>Artemisia annua</i> in <i>E. coli</i>. To apply to the iGEM standards, we removed an <i>EcoRI</i> site in the gene (AY835398.1).This gave us one of the basic parts we needed to create our system. </li><br/>
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<li> BBa_K1060011 is similar to BBa_K1060009. However, in this biobrick we added a <i>lac</i> operator in front of the beta farnesene synthase. This makes it possible to switch of (E)- β-farnesene production by using biosensors expressing LacI.</li><br />
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<li> <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1060009">BBa_K1060009</a> is a construct that constitutively expresses beta farnesene synthase. It is <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1060008">BBa_K1060008</a> behind a medium (constitutive) promoter and medium RBS (<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K608006">BBa_K608006</a>).<br/><b>Group favourite</b>: This generator will convert farnesyl pyrophosphate into the aphid alarm hormone, (E)-β-farnesene. This was the final device used for our <a href="http://2013.igem.org/Team:KU_Leuven/Project/Glucosemodel/EBF">Aphid experiments</a>.</li><br/>
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<li> <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1060011">BBa_K1060011</a> is similar to <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1060009">BBa_K1060009</a>. However, in this biobrick we added a <i>lac</i> operator in front of the beta farnesene synthase. This makes it possible to switch of (E)- β-farnesene production by using biosensors expressing LacI.<br/><b>Group favourite</b>: after the design of <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1060009">BBa_K1060009</a> this is the next step in the design of our <a href="http://2013.igem.org/Team:KU_Leuven/Project/Glucosemodel">glucose model</a>.</li><br/>
</ol>
</ol>
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   </html><groupparts>iGEM013 KU_Leuven</groupparts><html>
   </html><groupparts>iGEM013 KU_Leuven</groupparts><html>
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  </p>
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<ol>
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<h3>Other parts</h3>
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<li> <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1060000">BBa_K1060000</a> contains the aroG gene, coding for 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase and is a part of the Shikimate pathway. It converts phosphoenolpyruvate and erythrose-4P to chorismate. In our project chorismate was one of the precursors needed for the biosynthesis of <a href="http://2013.igem.org/Team:KU_Leuven/Project/Glucosemodel/MeS">methyl salicylate</a> (see also <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1060003">BBa_K1060003</a>). As basic chorismate levels inside <i>E. coli</i> are limited. We wanted to upregulate the biosynthesis of chorismate. The basic aroG gene can be repressed by phenylalanine. However, mutatations in the amino acid sequence (Pro150Leu and Leu175Asp) can prevent this repression by phenylalanine.</li><br/>
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 +
<li> <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1060001" target="_blank">BBa_K1060001</a> contains EBF synthase from <i>Streptomyces coelicolor</i>. In this organism the enzyme albaflavenone synthase (GenBank: AL939123.1) also has a second, completely distinct catalytic activity corresponding to the synthesis of farnesene isomers from farnesyl diphosphate (Zhao <i>et al.</i>, 2009). It results in the production of <a href="http://2013.igem.org/Team:KU_Leuven/Project/Glucosemodel/EBF" target="_blank">(E)-beta-farnesene</a> (61%).</li><br/>
 +
 
 +
<li> <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1060003">BBa_K1060003</a> is a generator for Methyl salicylate production. We requested the <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_J45700">BBa_J45700</a> brick from MIT 2006 and recloned into the pSB1C3 standard plasmid backbone. For the assembly of this part we refer to the original MIT brick <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_J45700">BBa_J45700</a> => "We assembled a complete wintergreen odor biosynthetic system (BBa_J45700) by combining the salicylic acid (<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_J45320" target="_blank">BBa_J45320</a>) and wintergreen odor generators (<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_J45120" target="_blank">BBa_J45120</a>). <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_J45700">BBa_J45700</a> catalyzes the conversion of the cellular metabolite chorismate to methyl salicylate or wintergreen odor."</li><br/>
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<li> <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1060004">BBa_K1060004</a> is a construction intermediate. It contains the BSMT1 enzyme (<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_J45120" target="_blank">BBa_J45120</a>) for the production of <a href="http://2013.igem.org/Team:KU_Leuven/Project/Glucosemodel/MeS">methyl salicylate</a> followed by a double terminator (<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_B0015" target="_blank">BBa_B0015</a>).</li><br/>
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 +
<li> <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1060005">BBa_K1060005</a> is a construction intermediate. It contains the pchBA enzyme (<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_J45320">BBa_J45320</a>) for the production of <a href="http://2013.igem.org/Team:KU_Leuven/Project/Glucosemodel/MeS>salicylate</a> followed a double terminator (<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_B0015">BBa_B0015</a>).</li><br/>
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 +
<li> <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1060006">BBa_K1060006</a> is an intermediate construct existing of the gene <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K082003" target="_blank">BBa_K082003</a> which codes for GFP (+degradation LVA tag), followed by the double terminator <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_B0015">BBa_B0015</a>. This is a part of our <a href="http://2013.igem.org/Team:KU_Leuven/Project/Oscillator/wetlab">feed-forward-loop</a>.</li><br/>
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 +
<li> <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1060007">BBa_K1060007</a> is an intermediate construct existing of the <i>cinI</i> <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_C0076"> BBa_C0076</a> gene followed by the double terminator <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_B0015">BBa_B0015</a>. <i>cinI</i> codes for an autoinducer synthetase and is derived from <i>Rizobium etli</i>. The quorum sensing molecule produced by this autoinducer synthetase is 3OH,C14:1-HSL. This construct was made to be incorporated in an <a href="http://2013.igem.org/Team:KU_Leuven/Project/Oscillator/wetlab"> oscillator</a>, that can be used for the alternating production of two different components. A quorum sensing molecule is necessary to get a colony-wide synchronization of oscillation. More information can be found on <a href="http://2013.igem.org/Team:KU_Leuven/Project/Oscillator">the oscillator page</a>.</li><br/>
 +
 
 +
<li> <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1060008">BBa_K1060008</a> is a construction intermediate in contains <a href="http://2013.igem.org/Team:KU_Leuven/Project/Glucosemodel/EBF">EBF synthase</a> (<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1060002">BBa_K1060002</a>) followed by the double terminator <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_B0015">BBa_B0015</a>.</li><br/>
 +
 
 +
<b>Note</b>: <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1060010">BBa_K1060010</a>, <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1060012">BBa_K1060012</a> and <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1060013">BBa_K1060013</a> are still under construction.
 +
 
  </div>
  </div>
</div>
</div>

Latest revision as of 21:45, 28 October 2013

iGem

Secret garden

Congratulations! You've found our secret garden! Follow the instructions below and win a great prize at the World jamboree!


  • A video shows that two of our team members are having great fun at our favourite company. Do you know the name of the second member that appears in the video?
  • For one of our models we had to do very extensive computations. To prevent our own computers from overheating and to keep the temperature in our iGEM room at a normal level, we used a supercomputer. Which centre maintains this supercomputer? (Dutch abbreviation)
  • We organised a symposium with a debate, some seminars and 2 iGEM project presentations. An iGEM team came all the way from the Netherlands to present their project. What is the name of their city?

Now put all of these in this URL:http://2013.igem.org/Team:KU_Leuven/(firstname)(abbreviation)(city), (loose the brackets and put everything in lowercase) and follow the very last instruction to get your special jamboree prize!

tree ladybugcartoon

Aphid Background

Crashcourse in aphid biology

Honeydew System

Our BanAphids react to honeydew

Sticker System

BanAphid population oscillates!

E. coligy

Validating the BanAphids in vivo

Parts

You are here!

Data Page

All our achievements on one small page!

Favourite parts

  1. BBa_K1060002 contains an open reading frame that codes for a sesquiterpene synthase (GenBank Accession No. AY835398), (E)-b-farnesene synthase, that has been isolated from Artemisia annua (Picaud et al., 2005). The enzyme converts farnesyl diphosphate into E-β-farnesene. E-β-farnesene is an alarm hormone produced by aphids. It is also an important semiochemical in aphid localization. It is released in the cornicle secretions of many aphid species (Franscis et al., 2005) to alert surrounding aphids of the presence of natural enemies (Kunert et al., 2005). Detection of these short range chemical cues leads not to the aphid directly, but only indicates the presence, improving prey detection of the predators and parasitoids.
    Group favourite: it was a milestone in our project work. We succeeded to clone the beta farnesene synthase of Artemisia annua in E. coli. To apply to the iGEM standards, we removed an EcoRI site in the gene (AY835398.1).This gave us one of the basic parts we needed to create our system.

  2. BBa_K1060009 is a construct that constitutively expresses beta farnesene synthase. It is BBa_K1060008 behind a medium (constitutive) promoter and medium RBS (BBa_K608006).
    Group favourite: This generator will convert farnesyl pyrophosphate into the aphid alarm hormone, (E)-β-farnesene. This was the final device used for our Aphid experiments.

  3. BBa_K1060011 is similar to BBa_K1060009. However, in this biobrick we added a lac operator in front of the beta farnesene synthase. This makes it possible to switch of (E)- β-farnesene production by using biosensors expressing LacI.
    Group favourite: after the design of BBa_K1060009 this is the next step in the design of our glucose model.

Loading.....

    Other parts

  1. BBa_K1060000 contains the aroG gene, coding for 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase and is a part of the Shikimate pathway. It converts phosphoenolpyruvate and erythrose-4P to chorismate. In our project chorismate was one of the precursors needed for the biosynthesis of methyl salicylate (see also BBa_K1060003). As basic chorismate levels inside E. coli are limited. We wanted to upregulate the biosynthesis of chorismate. The basic aroG gene can be repressed by phenylalanine. However, mutatations in the amino acid sequence (Pro150Leu and Leu175Asp) can prevent this repression by phenylalanine.

  2. BBa_K1060001 contains EBF synthase from Streptomyces coelicolor. In this organism the enzyme albaflavenone synthase (GenBank: AL939123.1) also has a second, completely distinct catalytic activity corresponding to the synthesis of farnesene isomers from farnesyl diphosphate (Zhao et al., 2009). It results in the production of (E)-beta-farnesene (61%).

  3. BBa_K1060003 is a generator for Methyl salicylate production. We requested the BBa_J45700 brick from MIT 2006 and recloned into the pSB1C3 standard plasmid backbone. For the assembly of this part we refer to the original MIT brick BBa_J45700 => "We assembled a complete wintergreen odor biosynthetic system (BBa_J45700) by combining the salicylic acid (BBa_J45320) and wintergreen odor generators (BBa_J45120). BBa_J45700 catalyzes the conversion of the cellular metabolite chorismate to methyl salicylate or wintergreen odor."

  4. BBa_K1060004 is a construction intermediate. It contains the BSMT1 enzyme (BBa_J45120) for the production of methyl salicylate followed by a double terminator (BBa_B0015).

  5. BBa_K1060005 is a construction intermediate. It contains the pchBA enzyme (BBa_J45320) for the production of BBa_B0015).

  6. BBa_K1060006 is an intermediate construct existing of the gene BBa_K082003 which codes for GFP (+degradation LVA tag), followed by the double terminator BBa_B0015. This is a part of our feed-forward-loop.

  7. BBa_K1060007 is an intermediate construct existing of the cinI BBa_C0076 gene followed by the double terminator BBa_B0015. cinI codes for an autoinducer synthetase and is derived from Rizobium etli. The quorum sensing molecule produced by this autoinducer synthetase is 3OH,C14:1-HSL. This construct was made to be incorporated in an oscillator, that can be used for the alternating production of two different components. A quorum sensing molecule is necessary to get a colony-wide synchronization of oscillation. More information can be found on the oscillator page.

  8. BBa_K1060008 is a construction intermediate in contains EBF synthase (BBa_K1060002) followed by the double terminator BBa_B0015.

  9. Note: BBa_K1060010, BBa_K1060012 and BBa_K1060013 are still under construction.