Team:Groningen/Project

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<div class="cte">
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<br>
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<div class="cte">
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<z1 >Abstract</z1><br>
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<div class="ctd">
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<z1>Abstract</z1>
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<td colspan="4">
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<p class="nomargin">
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Every year, one third of global food production -1.3 billion tons of food- is thrown away, partially due to the “best before” dating system.
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<z7>iGEM Groningen 2012</z7> seeks to provide an alternative method of assessing edibility: the <z7>Food Warden</z7>. It uses an <z7>engineered strain</z7>
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of <i>Bacillus subtilis</i> to detect and report volatiles in spoiling meat. The introduced <z7>genetic construct</z7> uses a promoter to trigger
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a pigment coding gene. This promoter, <z7>identified by microarray analysis</z7>, is significantly upregulated in the presence of
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<z7>volatiles from spoiling meat</z7>. The activity of the <z7>promoter</z7> regulates the expression of the <z7>pigment reporter</z7> and will
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be visible to the naked eye. For safe usage of the system, spores of our engineered strain are placed into one half of a semi-permeable
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<z7>capsule</z7>, the second containing a calibrated amount of nutrients. Breaking the barrier between the two compartments allows
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<z7>germination and growth</z7>, thereby activating the <z7>spoiling-meat sensor</z7>.
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</p>
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<img src="https://static.igem.org/mediawiki/2012/c/c2/Groningen2012_RR_20120909_rotting.png" width="100" height="100">
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<td align="left" >
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<td align="left" >
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<div class="cogoverlay" id="cogoverlaytopleft">
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<img src="https://static.igem.org/mediawiki/2012/1/15/Groningen2012_RR20120909_construct.png"  width="100px" height="100px">
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</div>
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</td>
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<td align="right" >
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<div class="cogoverlay" id="cogoverlaytopright">
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<img src="https://static.igem.org/mediawiki/2012/2/29/Groningen2012_RR20120909_badmeat.png" width="100px" height="100px">
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</div>
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</td>
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<td align="right">
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<div class="bigcog" id="bigcogbottomright">
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<img src="https://static.igem.org/mediawiki/2012/b/b6/Groningen2012_AD_20120802_BigCog.png" width="150px" height="150px">
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</div>
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</td>
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<td class="margincell">
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<div class="cogoverlay" id="cogoverlaybottomright">
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<img src="https://static.igem.org/mediawiki/2012/e/e7/Groningen2012_RR20120909_capsule.png" width="100px" height="100px">
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</td>
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</table>
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<br>
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<br>
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<div class="cte3">
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<div class="ctd3">
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<z2 >Completed after European Regionals</z2><br><br>
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</div>
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</div>
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<p class="marginChecklist">
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<br>
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<br>
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<a href="https://2012.igem.org/Team:Groningen/in_development" target="_blank"><img src="https://static.igem.org/mediawiki/2012/d/df/IGEMGroningen2012_AD20120915_Tick.png"> Tested a construct with downregulated promoter <i>WapA</i></a><br>
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<a href="https://2012.igem.org/Team:Groningen/Construct" target="_blank"><img src="https://static.igem.org/mediawiki/2012/d/df/IGEMGroningen2012_AD20120915_Tick.png"> Performed enhanced construct characterization</a><br>
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<a href="https://2012.igem.org/Team:Groningen/Sticker" target="_blank"><img src="https://static.igem.org/mediawiki/2012/d/df/IGEMGroningen2012_AD20120915_Tick.png"> Characterized the influence of oxygen on germination and growth within the sticker</a><br>
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<a href="https://2012.igem.org/Team:Groningen/market_research" target="_blank"><img src="https://static.igem.org/mediawiki/2012/d/df/IGEMGroningen2012_AD20120915_Tick.png"> Expanded the scope of the market survey</a><br>
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<a href="https://2012.igem.org/Team:Groningen/international_cooperation" target="_blank"><img src="https://static.igem.org/mediawiki/2012/d/df/IGEMGroningen2012_AD20120915_Tick.png"> Collaborated with Cambridge 2012</a><br>
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<br>
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</p>
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<a name="MainAcc"></a>
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<div class="cte2">
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<div class="ctd2">
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<a name="MainAcc"></a><z2 >Our main accomplishments</z2><br><br>
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</div>
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</div>
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<br>
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<br>
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<p align=center style="color: white; font-size: 10pt;">
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<i>Hover your mouse over the pictures to see more!</i>
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</p>
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<img src="https://static.igem.org/mediawiki/2012/c/cb/Groningen2012_RR1capsule_break.png" width=200 />
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<a href="#"><img src="https://static.igem.org/mediawiki/2012/c/c2/Groningen2012_RR_20120909_rotting.png" width=150 /><img src="https://static.igem.org/mediawiki/2012/7/74/Groningen2012_ADVolatilesBig.png" class="preview" width=400 /></a>
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<font color=#FF6700><br><b>1.</b></font> We designed and tested the "<a class="inlink" href="https://2012.igem.org/Team:Groningen/Sticker">Sticker</a>": a capsule in which bacteria are kept inside and volatiles can go through. We used a model to get insight on how to tweak the growth of <i>Bacillus subtilis</i>.
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<font color=#FF6700><b>2.</b></font> We explored the definition of spoiled meat and ways to check meat spoilage. We identified <a class="inlink" href="https://2012.igem.org/Team:Groningen/volatiles">various compounds</a> present in spoiled meat.<br>
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<img src="https://static.igem.org/mediawiki/2012/2/29/Groningen2012_RR20120909_badmeat.png" width=200 />
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                        color: #FFF;
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<img src="https://static.igem.org/mediawiki/2012/e/ec/Groningen2012_ADSensorBig.png" class="preview" width=400 />
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<stickerP ><A HREF="https://2012.igem.org/Team:Groningen/Sticker" TARGET="_blank"><FONT COLOR=#ffffff>sticker</FONT></A></stickerP>
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<li>
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<a href="#">
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<img src="https://static.igem.org/mediawiki/2012/2/24/Groningen2012_RRADPsac_transp.png" width=150 />
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<img src="https://static.igem.org/mediawiki/2012/c/c1/Groningen2012_AdBackboneBig.png" class="preview" width=400 />
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</a>
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<font color=#FF6700><b>3.</b></font> We identified spoiled meat <a class="inlink" href="https://2012.igem.org/Team:Groningen/Sensor">sensors</a> by transcriptome analysis.<br><br><br>
 +
</p>
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</td>
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<td class="accCap">
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<p class="small">
 +
<font color=#FF6700><b>4.</b></font> <a class="inlink" href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K818000" target="_blank">Backbone pSac-Cm</a>: easy cloning in<br><i>B. subtilis</i>, the BioBrick way, for the first time! It's easy to check, BioBrick compatible, <i>E. coli</i> compatible, stabily inserted into the <i>B. subtilis</i> chromosome.
 +
</p>
 +
</td>
 +
</tr>
 +
<tr>
 +
<td class="accPic">
 +
<ul class="hoverbox">
 +
<li>
 +
<a href="#">
 +
<img src="https://static.igem.org/mediawiki/2012/f/f1/Groningen2012_RRADPigments_transp.png" width=150 />
 +
<img src="https://static.igem.org/mediawiki/2012/3/35/Groningen2012_ADPigmentsBig.png"  class="preview" width=400 />
 +
</a>
 +
</li>
 +
</ul>
 +
</td>
 +
<td class="accPic">
 +
<ul class="hoverbox" style="margin-left: 30px">
 +
<li>
 +
<a href="#">
 +
<img src="https://static.igem.org/mediawiki/2012/9/93/Groningen2012_ADConstruct_yellow.png" width=220 />
 +
<img src="https://static.igem.org/mediawiki/2012/2/21/Groningen2012_ADConstructBig.png" class="preview" width=400 />
 +
</a>
 +
</li>
 +
</ul>
 +
</td>
 +
</tr>
 +
<tr>
 +
<td class="accCap">
 +
<p class="small">
 +
<font color=#FF6700><br><br><b>5.</b></font> We made <a class="inlink" href="https://2012.igem.org/Team:Groningen/pigmentproduction">AmilCP and AmilGFP</a> suitable for expression in <i>Bacillus subtilis</i>.
 +
<br>
 +
These chromoproteins can be of significant value to the other <i>Bacillus subtilis</i> users in the BioBrick community.
 +
<br><br><br>
 +
</p>
 +
</td>
 +
<td class="accCap">
 +
<p class="small">
 +
<font color=#FF6700><b>6.</b></font> Most importantly: we developed a <a class="inlink" href="https://2012.igem.org/Team:Groningen/Construct">construct</a> which makes <i>Bacillus subtilis</i> sense spoiled meat and produce an output in the form of a yellow or purple pigment visible by naked eye.
 +
<br><br><br>
 +
</p>
 +
</td>
 +
</tr>
 +
</table>
 +
<br>
 +
<br>
 +
<br>
 +
</body>
</html>
</html>
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</style>
 
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<div class="Germination2">
 
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<div class="Germination1">
 
-
<GerminationP ><A HREF="https://2012.igem.org/Team:Groningen/Germination" TARGET="_blank"><FONT COLOR=#ffffff>germination</FONT></A></GerminationP>
 
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font-family: Helvetica, Arial, sans-serif;
 
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                        background: #000;
 
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padding-right:10px;
 
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padding-left:10px;
 
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background-color: rgba(0,0,0,0.3);
 
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                        color: #FFF;
 
-
 
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}
 
-
</style>
 
-
 
-
<div class="PBADmeat12">
 
-
<div class="PBADmeat11">
 
-
<PBADmeat1P ><A HREF="https://2012.igem.org/Team:Groningen/identifyingbadvolatiles" TARGET="_blank"><FONT COLOR=#ffffff>volatiles</FONT></A></PBADmeat1>
 
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</div>
 
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</div>
 
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</body>
 
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</head>
 
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</html>
 
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background-color: rgba(0,0,0,0.3);
 
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padding-right:10px;
 
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padding-left:10px;
 
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background-color: transparent;
 
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background-color: rgba(0,0,0,0.3);
 
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                        color: #FFF;
 
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-
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-
</style>
 
-
 
-
<div class="identification2">
 
-
<div class="identification1">
 
-
<identificationP ><A HREF="https://2012.igem.org/Team:Groningen/identifyingPBADmeat" TARGET="_blank"><FONT COLOR=#ffffff>identification</FONT></A></identificationP>
 
-
</div>
 
-
</div>
 
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</body>
 
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</head>
 
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</html>
 
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<style>
 
-
 
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body {
 
-
font-family: Helvetica, Arial, sans-serif;
 
-
 
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                }
 
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PigmentsP {
 
-
font-size:12pt;
 
-
                        line-height:12pt;
 
-
                        color:white;
 
-
}
 
-
                .Pigments1 {
 
-
                        width: 100px;
 
-
text-align: center;
 
-
                        background: #000;
 
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background-color: transparent;
 
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background-color: rgba(0,0,0,0.3);
 
-
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-
                        margin: 0px auto;
 
-
                        color: #FFF;
 
-
}
 
-
                .Pigments2 {
 
-
left: 350px;
 
-
top:520px;
 
-
position:absolute;
 
-
                        width: 120px;
 
-
text-align: center;
 
-
                        background: #000;
 
-
padding-right:10px;
 
-
padding-left:10px;
 
-
                        margin: 0 auto;
 
-
background-color: transparent;
 
-
background-color: rgba(0,0,0,0.3);
 
-
                        color: #FFF;
 
-
 
-
}
 
-
</style>
 
-
 
-
<div class="Pigments2">
 
-
<div class="Pigments1">
 
-
<PigmentsP ><A HREF="https://2012.igem.org/Team:Groningen/Pigments" TARGET="_blank"><FONT COLOR=#ffffff>pigment</FONT></A></Pigments>
 
-
</div>
 
-
</div>
 
-
</body>
 
-
</head>
 
-
</html>
 
-
 
-
 
-
<html>
 
-
<head>
 
-
<style>
 
-
 
-
body {
 
-
font-family: Helvetica, Arial, sans-serif;
 
-
 
-
                }
 
-
ConstructP {
 
-
font-size:12pt;
 
-
                        line-height:12pt;
 
-
                        color:white;
 
-
}
 
-
                .Construct1 {
 
-
                        width: 100px;
 
-
text-align: center;
 
-
                        background: #000;
 
-
background-color: transparent;
 
-
background-color: rgba(0,0,0,0.3);
 
-
                        padding: 10px ;
 
-
                        margin: 0px auto;
 
-
                        color: #FFF;
 
-
}
 
-
                .Construct2 {
 
-
left: 530px;
 
-
top:520px;
 
-
position:absolute;
 
-
                        width: 120px;
 
-
text-align: center;
 
-
                        background: #000;
 
-
padding-right:10px;
 
-
padding-left:10px;
 
-
                        margin: 0 auto;
 
-
background-color: transparent;
 
-
background-color: rgba(0,0,0,0.3);
 
-
                        color: #FFF;
 
-
 
-
}
 
-
</style>
 
-
 
-
<div class="Construct2">
 
-
<div class="Construct1">
 
-
<ConstructP ><A HREF="https://2012.igem.org/Team:Groningen/Construct" TARGET="_blank"><FONT COLOR=#ffffff>construct</FONT></A></ConstructP>
 
-
</div>
 
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-
</body>
 
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</head>
 
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</html>
 
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<html>
 
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<head>
 
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<style>
 
-
 
-
body {
 
-
font-family: Helvetica, Arial, sans-serif;
 
-
 
-
                }
 
-
DeathP {
 
-
font-size:12pt;
 
-
                        line-height:12pt;
 
-
                        color:white;
 
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}
 
-
                .Death1 {
 
-
                        width: 100px;
 
-
text-align: center;
 
-
                        background: #000;
 
-
background-color: transparent;
 
-
background-color: rgba(0,0,0,0.3);
 
-
                        padding: 10px ;
 
-
                        margin: 0px auto;
 
-
                        color: #FFF;
 
-
}
 
-
                .Death2 {
 
-
left: 440px;
 
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top:560px;
 
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position:absolute;
 
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                        width: 120px;
 
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text-align: center;
 
-
                        background: #000;
 
-
padding-right:10px;
 
-
padding-left:10px;
 
-
                        margin: 0 auto;
 
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background-color: transparent;
 
-
background-color: rgba(0,0,0,0.3);
 
-
                        color: #FFF;
 
-
 
-
}
 
-
</style>
 
-
 
-
<div class="Death2">
 
-
<div class="Death1">
 
-
<DeathP ><A HREF="https://2012.igem.org/Team:Groningen/Death" TARGET="_blank"><FONT COLOR=#ffffff>kill switch</FONT></A></DeathP>
 
-
</div>
 
-
</div>
 
-
</body>
 
-
</head>
 
-
</html>
 
-
 
-
 
-
 
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-
<html>
 
-
<div style="position:absolute; left: 80px; top: 500px;opacity:0.4; filter:alpha(opacity=40);z-index:-1;">
 
-
<img src="https://static.igem.org/mediawiki/2012/b/b6/Groningen2012_AD_20120802_BigCog.png" width="150" height="150">
 
-
</div>
 
-
 
-
<div style="position:absolute; left: 100px; top: 510px;">
 
-
<img src="https://static.igem.org/mediawiki/2012/c/c2/Groningen2012_RR_20120909_rotting.png" width="100" height="100">
 
-
</div>
 
-
 
-
<div style="position:absolute; right: 80px; top: 500px;opacity:0.4; filter:alpha(opacity=40); z-index:-1;">
 
-
<img src="https://static.igem.org/mediawiki/2012/b/b6/Groningen2012_AD_20120802_BigCog.png" width="150" height="150">
 
-
</div>
 
-
 
-
<div style="position:absolute; right: 110px; top: 525px;">
 
-
<img src="https://static.igem.org/mediawiki/2012/e/e7/Groningen2012_RR20120909_capsule.png" width="100" height="100">
 
-
</div>
 
-
 
-
<div style="position:absolute; left: 0px; top: 300px;opacity:0.4; filter:alpha(opacity=40); z-index:-1;">
 
-
<img src="https://static.igem.org/mediawiki/2012/b/b6/Groningen2012_AD_20120802_BigCog.png" width="150" height="150">
 
-
</div>
 
-
 
-
<div style="position:absolute; right: 20px; top: 320px;">
 
-
<img src="https://static.igem.org/mediawiki/2012/2/29/Groningen2012_RR20120909_badmeat.png" width="100" height="100">
 
-
</div>
 
-
 
-
<div style="position:absolute; right: 0px; top: 300px;opacity:0.4; filter:alpha(opacity=40); z-index:-1;">
 
-
<img src="https://static.igem.org/mediawiki/2012/b/b6/Groningen2012_AD_20120802_BigCog.png" width="150" height="150">
 
-
</div>
 
-
 
-
<div style="position:absolute; left: 20px; top: 330px;">
 
-
<img src="https://static.igem.org/mediawiki/2012/1/15/Groningen2012_RR20120909_construct.png" width="100" height="100">
 
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</div>
 
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</body>
 
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</html>
 
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<html>
 
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<head>
 
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<style type="text/css">
 
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p.marginleft
 
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{
 
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margin-top:-150px;
 
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margin-left:150px;
 
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font-size:13pt;
 
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line-height:15pt;
 
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margin-right:150px;
 
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color:white;
 
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{
 
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margin-right:150px;
 
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-
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margin-bottom:0px;
 
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font-size:18pt;
 
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line-height:21pt;
 
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color:white;
 
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}
 
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                .ctd2 {
 
-
                        width: 300px;
 
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                        height: 20px;
 
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                        text-align: center;
 
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                        background: #000;
 
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                        background-color: transparent;
 
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                        background-color: rgba(0,0,0,0.3);
 
-
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-
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                        padding-left:10px;
 
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<p class="marginleft">
 
-
Every year, one third of global food production -1.3 billion tons of food- is thrown away, partially due to the “best before” dating system. <font color=#FF6700><b>iGEM Groningen 2012</b></font> seeks to provide an alternative method of assessing edibility: the <font color=#FF6700><b>Food Warden</b></font>. It uses an <font color=#FF6700><b>engineered strain </b></font> of <i>Bacillus subtilis</i> to detect and report volatiles in spoiling meat. The introduced <font color=#FF6700><b>genetic construct</b></font> uses a promoter to trigger a pigment coding gene. This promoter, <font color=#FF6700><b>identified by microarray analysis</b></font>, is significantly up-regulated in the presence of <font color=#FF6700><b>volatiles from spoiling meat</b></font>. The activity of the <font color=#FF6700><b>promoter</b></font> regulates the expression of the <font color=#FF6700><b>pigment reporter</b></font> and will be visible to the naked eye. For safe usage of the system, spores of our engineered strain are placed into one half of a semi-permeable <font color=#FF6700><b>capsule</b></font>, the second containing a calibrated amount of nutrients. Breaking the barrier between the two compartments allows <font color=#FF6700><b>germination and growth</b></font>, thereby activating the <font color=#FF6700><b>spoiling meat sensor</b></font>.<br><br>
 
-
 
-
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-
<br><br><br><br><br><br><br><br><br><br><br><br><br><br><br>
 
-
<div class="cte2">
 
-
<div class="ctd2">
 
-
<z2 >Our main accomptishments</z2><br>
 
-
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<p class=orange>In the lab</p>
 
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{{Template:SponsorsGroningen2012}}
{{Template:SponsorsGroningen2012}}

Latest revision as of 02:00, 27 October 2012





Abstract

Every year, one third of global food production -1.3 billion tons of food- is thrown away, partially due to the “best before” dating system. iGEM Groningen 2012 seeks to provide an alternative method of assessing edibility: the Food Warden. It uses an engineered strain of Bacillus subtilis to detect and report volatiles in spoiling meat. The introduced genetic construct uses a promoter to trigger a pigment coding gene. This promoter, identified by microarray analysis, is significantly upregulated in the presence of volatiles from spoiling meat. The activity of the promoter regulates the expression of the pigment reporter and will be visible to the naked eye. For safe usage of the system, spores of our engineered strain are placed into one half of a semi-permeable capsule, the second containing a calibrated amount of nutrients. Breaking the barrier between the two compartments allows germination and growth, thereby activating the spoiling-meat sensor.



Completed after European Regionals



Tested a construct with downregulated promoter WapA
Performed enhanced construct characterization
Characterized the influence of oxygen on germination and growth within the sticker
Expanded the scope of the market survey
Collaborated with Cambridge 2012

Our main accomplishments



Hover your mouse over the pictures to see more!



1. We designed and tested the "Sticker": a capsule in which bacteria are kept inside and volatiles can go through. We used a model to get insight on how to tweak the growth of Bacillus subtilis.

2. We explored the definition of spoiled meat and ways to check meat spoilage. We identified various compounds present in spoiled meat.

3. We identified spoiled meat sensors by transcriptome analysis.

4. Backbone pSac-Cm: easy cloning in
B. subtilis, the BioBrick way, for the first time! It's easy to check, BioBrick compatible, E. coli compatible, stabily inserted into the B. subtilis chromosome.



5. We made AmilCP and AmilGFP suitable for expression in Bacillus subtilis.
These chromoproteins can be of significant value to the other Bacillus subtilis users in the BioBrick community.


6. Most importantly: we developed a construct which makes Bacillus subtilis sense spoiled meat and produce an output in the form of a yellow or purple pigment visible by naked eye.





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