Team:Groningen/DesignTest2

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<z1>Attributions</z1>
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<z1>Abstract</z1>
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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
 +
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
 +
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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<img src="https://static.igem.org/mediawiki/2012/1/15/Groningen2012_RR20120909_construct.png"  width="100px" height="100px">
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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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<br>
<br>
-
<br>
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<br>
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<p>
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<div class="cte2">
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All the work for this project was performed by the iGEM Groningen 2012 team members, the lab work starting from May 2012.
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<div class="ctd2">
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<br>
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<a name="MainAcc"></a><z2 >Our main accomplishments</z2><br>
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<br>
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</div>
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The original <i>Bacillus subtilis</i> integration vector psac-cm from which we derived our backbone biobrick was provided
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</div>
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by Dr. Jan-Willem Veening from the Molecular Genetics group of the University of Groningen, just as the GFP optimized for
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<br><br>
-
<i>Bacillus subtilis</i> for our testing constructs. Our <i>Escherichia coli dh5a</i> and <i>Bacillus subtilis sp. 168</i>
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<p class=orange>In the lab</p>
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were provided by Dr. Jan-Willem Veening and MolGen.
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<p class="marginleft">
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<br>
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<img src="https://static.igem.org/mediawiki/2012/d/df/IGEMGroningen2012_AD20120915_Tick.png" width=20> Most importantly: we developed a construct which makes <i>Bacillus subtilis</i> sense spoiled meat and produce an output in the form of a yellow pigment visible by naked eye.<br><br>
-
<br>
+
<img src="https://static.igem.org/mediawiki/2012/d/df/IGEMGroningen2012_AD20120915_Tick.png" width=20> Design of the “<a href="https://2012.igem.org/Team:Groningen/Sticker">Sticker</a>”: semi-permeable capsule: bacteria are kept inside, volatiles can go through. Proof that <i>Bacillus subtilis</i> grows inside the sticker.<br><br>
-
The idea of the Food Warden was developed by the team before the labwork started. All experiments, the modeling and the
+
<img src="https://static.igem.org/mediawiki/2012/d/df/IGEMGroningen2012_AD20120915_Tick.png" width=20> Development of BioBrick psac-cm for <i>Bacillus subtilis</i>. <br><a href="http://partsregistry.org/Part:BBa_K818000" target=_blank><font size=4 color=#FF6700><b>Easy cloning in <i>Bacillus subtilis</i> in Biobrick fashion for the first time!</b></font></a> Advantages: easy to check, BioBrick compatible, <i>E. coli</i> compatible, stable insertion into <i>Bacillus subtilis</i> chromosome.<br><br>
-
sticker design was planned and carried out by the team. Supervision and direction was given by our supervisors, but they
+
<img src="https://static.igem.org/mediawiki/2012/d/df/IGEMGroningen2012_AD20120915_Tick.png" width=20> Identification of spoiled meat sensors by transcriptome analysis. Usage of an innovative experimental setup which can be used to test more sensors in the future.<br><br>
-
only had a consultative function.
+
<img src="https://static.igem.org/mediawiki/2012/d/df/IGEMGroningen2012_AD20120915_Tick.png" width=20> We made AmilCP and AmilGFP be expressed in <i>Bacillus subtilis</i>. These chromoproteins can be of significant value to the other <i>Bacillus subtilis</i> users in the BioBrick community.<br><br><br>
-
<br>
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</p>
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<br>
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<p class=orange>Other</p>
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We were informed by Marius Uebel about GMP/SOP and data management with the use of acronyms. The team also took care of
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<p class="marginleft">
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the sponsoring and all the other organizational issues. We made our own logos, drawings and wiki/presentation/poster style.  
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<img src="https://static.igem.org/mediawiki/2012/d/df/IGEMGroningen2012_AD20120915_Tick.png" width=20> Smart spores activation system: inspired by "glow in the dark stick", we designed a sticker with two compartments. One compartment holds the spores, the other the nutrients; when the nutrients compartment is braked, the spores start to germinate, activating the sticker.<br><br>
-
The picture gallery on the home page has been made on the basis of the hoverbox image gallery code by
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<img src="https://static.igem.org/mediawiki/2012/d/df/IGEMGroningen2012_AD20120915_Tick.png" width=20> We explored the definition of spoiled meat and different ways to test this.<br><br>
-
<a class="inlink" href="http://sonspring.com/journal/hoverbox-image-gallery">Nathan Smith</a>.
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<img src="https://static.igem.org/mediawiki/2012/d/df/IGEMGroningen2012_AD20120915_Tick.png" width=20> We identified various compounds present in spoiled meat by Gas Chromatography - Mass Spectrometry.<br><br>
-
<br>
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<img src="https://static.igem.org/mediawiki/2012/d/df/IGEMGroningen2012_AD20120915_Tick.png" width=20> We made a model which gives insight on how to tweak the growth of <i>Bacillus subtilis</i> based upon literature, experimental data and flux balance analysis.<br>
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<br>
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-
<br>
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-
<br>
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{{Template:SponsorsGroningen2012}}
{{Template:SponsorsGroningen2012}}

Latest revision as of 00:16, 27 September 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.



Our main accomplishments


In the lab

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

Design of the “Sticker”: semi-permeable capsule: bacteria are kept inside, volatiles can go through. Proof that Bacillus subtilis grows inside the sticker.

Development of BioBrick psac-cm for Bacillus subtilis.
Easy cloning in Bacillus subtilis in Biobrick fashion for the first time! Advantages: easy to check, BioBrick compatible, E. coli compatible, stable insertion into Bacillus subtilis chromosome.

Identification of spoiled meat sensors by transcriptome analysis. Usage of an innovative experimental setup which can be used to test more sensors in the future.

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


Other

Smart spores activation system: inspired by "glow in the dark stick", we designed a sticker with two compartments. One compartment holds the spores, the other the nutrients; when the nutrients compartment is braked, the spores start to germinate, activating the sticker.

We explored the definition of spoiled meat and different ways to test this.

We identified various compounds present in spoiled meat by Gas Chromatography - Mass Spectrometry.

We made a model which gives insight on how to tweak the growth of Bacillus subtilis based upon literature, experimental data and flux balance analysis.


Our sponsors:

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