Difference between revisions of "Team:ITB Indonesia/Design"

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{{ITB_Indonesia}}
 
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<h1>Design</h1>
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<p>
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Design is the first step in the design-build-test cycle in engineering and synthetic biology. Use this page to describe the process that you used in the design of your parts. You should clearly explain the engineering principles used to design your project.
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This page is different to the "Applied Design Award" page. Please see the <a href="https://2017.igem.org/Team:ITB_Indonesia/Applied_Design">Applied Design</a> page for more information on how to compete for that award.
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<div class="column full_size" style="background: #1c2922; margin-top: -40px; padding: 30px; color: #e8e6d1">
<h5>What should this page contain?</h5>
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<h1 class="ITB_h1">Design</h1>
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<li>Explanation of the engineering principles your team used in your design</li>
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<li>Discussion of the design iterations your team went through</li>
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<li>Experimental plan to test your designs</li>
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<div style="background: #e8e6d1; padding: 30px; color: #1c2922">
<h5>Inspiration</h5>
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<h1 class="ITB_h1" style="padding-bottom: 30px; margin-bottom: 50px; border-bottom: 2px solid #1c2922 !important; padding-left: 30px; text-align: center; color: #1c2922">Design</h1>
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<li><a href="https://2016.igem.org/Team:MIT/Experiments/Promoters">2016 MIT</a></li>
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<li><a href="https://2016.igem.org/Team:BostonU/Proof">2016 BostonU</a></li>
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<li><a href="https://2016.igem.org/Team:NCTU_Formosa/Design">2016 NCTU Formosa</a></li>
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<p>We designed <i>E. coli</i> that have increased biofilm formation speed compared to wild-type by inserting NhaR transcriptional activator. Yet, both of them are also inserted by PETase, a protein that will degrade PET plastic into its smaller monomers.</p>
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<p>The production and degradation capacity are modeled to predict when degradation does not occur or reaction rate equals to zero. Hence, we want to test following hypothesis, whether the increased expression of biofilm will support the PET degradation capacity. Our assumptions are as follows: (1) Biofilm which encapsulates <i>E. coli</i> colony act as protection from environment and increase colony concentration inside biofilm, therefore enzyme production will be optimal. (2) Bottom section of <i>E. coli</i> that interacts with the PET interface is not encapsulated by biofilm and the enzyme-substrate complex will work just as fine on this layer.</p>
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<p><i>E. coli</i> without increased biofilm formation is used as comparison to above strain. Without NhaR, this strain will just have normal biofilm formation expression, which will have planktonic forms floating around in the medium. PET degradation will be slower than using <i>+NhaR</i> strain, because we assume that PETase should be diffused into water and affected by Brownian motion, therefore the possibility of PETase-substrate contact will be less than <i>+NhaR</i> strain. </p>
  
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<p>If, somehow, the result of <i>+NhaR</i> strain is significantly lower than its opposite strain, we should conclude that our assumptions is not confirmed to be true.</p>
  
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<center><figure>
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    <img src='https://static.igem.org/mediawiki/2017/b/b8/Design_itbIndonesia.JPG'/>
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    <figcaption><i><b>Figure 1:</b> Left: assumed process of +NhaR strain. Right: assumed process of -NhaR strain</i></figcaption>
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Latest revision as of 14:50, 1 November 2017


Design


Design

We designed E. coli that have increased biofilm formation speed compared to wild-type by inserting NhaR transcriptional activator. Yet, both of them are also inserted by PETase, a protein that will degrade PET plastic into its smaller monomers.

The production and degradation capacity are modeled to predict when degradation does not occur or reaction rate equals to zero. Hence, we want to test following hypothesis, whether the increased expression of biofilm will support the PET degradation capacity. Our assumptions are as follows: (1) Biofilm which encapsulates E. coli colony act as protection from environment and increase colony concentration inside biofilm, therefore enzyme production will be optimal. (2) Bottom section of E. coli that interacts with the PET interface is not encapsulated by biofilm and the enzyme-substrate complex will work just as fine on this layer.

E. coli without increased biofilm formation is used as comparison to above strain. Without NhaR, this strain will just have normal biofilm formation expression, which will have planktonic forms floating around in the medium. PET degradation will be slower than using +NhaR strain, because we assume that PETase should be diffused into water and affected by Brownian motion, therefore the possibility of PETase-substrate contact will be less than +NhaR strain.

If, somehow, the result of +NhaR strain is significantly lower than its opposite strain, we should conclude that our assumptions is not confirmed to be true.

Figure 1: Left: assumed process of +NhaR strain. Right: assumed process of -NhaR strain