Difference between revisions of "Team:ASIJ Tokyo/Parts"

 
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         <li class="active"><a href="https://2016.igem.org/Team:ASIJ_Tokyo">Home</a></li>
 
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           <a class="dropdown-toggle" data-toggle="dropdown" href="https://2016.igem.org/ASIJProjectDescription">Our Project<span class="caret"></span></a>
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           <a class="dropdown-toggle" data-toggle="dropdown" href="">Project<span class="caret"></span></a>
 
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             <li><a href="#">Page 1-1</a></li>
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             <li><a href="https://2016.igem.org/Team:ASIJ_Tokyo/Description">Project Description + Abstract</a></li>
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             <li><a href="#">Page 1-2</a></li>
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             <li><a href="https://2016.igem.org/Team:ASIJ_Tokyo/Experiments">Experiments</a></li>
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             <li><a href="#">Page 1-3</a></li>
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             <li><a href="https://2016.igem.org/Team:ASIJ_Tokyo/Results">Results</a></li>
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            <li><a href="https://2016.igem.org/Team:ASIJ_Tokyo/Notebook">Notebook</a></li>
 
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         <li><a href="https://2016.igem.org/Team:ASIJ_Tokyo/Team">Team</a></li>
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         <li><a href="https://2016.igem.org/ASIJParts">Parts</a></li>
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         <li><a href="https://2016.igem.org/Team:ASIJ_Tokyo/Safety">Safety</a></li>
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         <li><a href="https://2016.igem.org/ASIJPROTOCOL">Protocol</a></li>
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         <li><a href="https://2016.igem.org/Team:ASIJ_Tokyo/Attributions">Attributions</a></li>
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         <li><a href="#">Pictures</a></li>
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         <li><a href="https://2016.igem.org/Team:ASIJ_Tokyo/Parts">Parts</a></li>    
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         <li><a href="https://2016.igem.org/Team:ASIJ_Tokyo/Human_Practices">Human Practices</a></li>
 
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                   <h1>Parts </h1>
 
                   <h1>Parts </h1>
 
<br>
 
<br>
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<ol type="1">
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<h3><strong>Anderson Promoters</strong></h3>
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<li> Weak promoter: Wanted a weak promoter to measure its PET plastic degradation up against the other promoters
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<li>Relative Strength: weak (0.33) BBa_J23110
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<li>Moderate promoter: Wanted a moderate promoter to measure its PET plastic degradation up against the other promoters
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        <ul>
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<li>Strong promoter: Wanted a strong promoter to measure its PET plastic degradation up against the other promoters
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            <li>Selected as a relatively weak promoter against which to compare the expression of the PETase construct</li>
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<li>N-Osmy tag: Fusion proteins with an N-terminal osmY have been shown to successfully secrete proteins of interest
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          </ul>
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<li>C-Myc tag: Make our PETase construct easier to detect for Western Blot to assay expression and secretion
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<li>PETase sequence: Needed the enzyme to degrade PET plastic
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−
<li>Plasmid backbone: pSB1C3: Best vector to move our ligated DNA produced in Gibson Assembly
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<li>High efficiency e. Coli cells: Were the host for our optimized PETase
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</li>
 
</li>
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</ol>
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<li>Relative Strength: moderate (0.58) BBa_J23111
 +
          <ul>
 +
            <li>Selected as a relatively moderately strong promoter against which to compare the expression of the PETase construct </li>
 +
          </ul>
 +
</li>
 +
<li>Relative Strength: strong (1) BBa_J23100
 +
          <ul>
 +
            <li>Selected as a relatively strong promoter against which to compare the expression of the PETase construct</li>
 +
          </ul>
 +
</li>
 +
<br>
 +
<h3><strong>Tags</strong></h3>
 +
<li>N-OsmY
 +
      <ul>
 +
          <li>Fusion proteins with an N-terminal osmY have been shown to successfully secrete proteins of interest out of <i>E. Coli</i></li>
 +
      </ul>
 +
</li>
 +
<li>C-Myc
 +
    <ul>
 +
      <li>This C-terminal tag would allow the PETase construct to be more easily detected by Western blot assays for expression </li>
 +
    </ul>
 +
</li>
 +
<br>
 +
<h3><strong>Others</strong></h3>
 +
<li>PETase sequence
 +
<ul>
 +
      <li>Gene that coded for the PET hydrolase</li>
 +
</ul>
 +
</li>
 +
<li>Plasmid backbone
 +
<ul>
 +
    <li>The pSB1C3 backbone is recommended</li>
 +
</ul>
 +
</li>
 +
<li>High efficiency <i>E. Coli</i> cells
 +
<ul>
 +
    <li>Hosted the PETase constructs</li>
 +
</ul>
 +
</li>
 +
<br>
 +
<h3><strong>Parts Submitted</strong></h3>
 +
<li>Gibson-assembled pSB1C3 with novel fusion osmY-PETase A1 (BBa_K1885001)
 +
    <ul>
 +
          <li>This part offers efficiency in the initial step in the bioremediation of polyethylene terephthalate (PET): the production of metabolizable monomers (ethylene glycol and terephthalate) through hydrolysis. PETase catalyzes the hydrolysis of PET into terephthalate and ethylene glycol monomers. The N-terminal osmY protein and its signal peptide fused to PETase, with serine-glycine peptide’s repeating intervention, serves to secrete PETase out of the cell (this reacts with PET in the cell's local environment). The Anderson promoter, with a relative strength of 1 that precedes the fusion protein in the plasmid, serves to constitutively express the fusion PETase, enhancing the degradation of environmental PET. This part utilizes the pSB1C3 backbone and confers resistance to chloramphenicol.</li>
 +
    </ul>
 +
</li>
 +
<li>Gibson-assembled pSB1C3 with novel fusion osmY-PETase A0.58 (BBa_K1885002)
 +
    <ul>
 +
          <li>This part offers efficiency in the initial step in the bioremediation of polyethylene terephthalate (PET): the production of metabolizable monomers (ethylene glycol and terephthalate) through hydrolysis. PETase catalyzes the hydrolysis of PET into terephthalate and ethylene glycol monomers. The N-terminal osmY protein and its signal peptide fused to PETase, with serine-glycine peptide’s repeating intervention, serves to secrete PETase out of the cell (this reacts with PET in the cell's local environment). The Anderson promoter, with a relative strength of 0.58 that precedes the fusion protein in the plasmid, serves to constitutively express the fusion PETase, enhancing the degradation of environmental PET. This part utilizes the pSB1C3 backbone and confers resistance to chloramphenicol.</li>
 +
    </ul>
 +
</li>
 +
<li>Gibson-assembled pSB1C3 with novel fusion osmY-PETase A0.33 (BBa_K1885005)
 +
    <ul>
 +
          <li>This part offers efficiency in the initial step in the bioremediation of polyethylene terephthalate (PET): the production of metabolizable monomers (ethylene glycol and terephthalate) through hydrolysis. PETase catalyzes the hydrolysis of PET into terephthalate and ethylene glycol monomers. The N-terminal osmY protein and its signal peptide fused to PETase, with serine-glycine peptide’s repeating intervention, serves to secrete PETase out of the cell (this reacts with PET in the cell's local environment). The Anderson promoter, with a relative strength of 0.33 that precedes the fusion protein in the plasmid, serves to constitutively express the fusion PETase, enhancing the degradation of environmental PET. This part utilizes the pSB1C3 backbone and confers resistance to chloramphenicol.</li>
 +
    </ul>
 +
</li>
 +
 
 
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<script src="https://ajax.googleapis.com/ajax/libs/jquery/1.12.4/jquery.min.js"></script>
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    <!--CORE SCRIPTS-->
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    <script src="assets/js/bootstrap.js"></script>
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    <!--WOW SCRIPTS-->
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    <script src="assets/js/wow.js"></script>
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    <!--SCROLL SCRIPTS-->
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    <script src="assets/js/jquery.smooth-scroll.min.js"></script>
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Latest revision as of 23:32, 19 October 2016

The BIG TEMPLATE : RESPONSIVE and FREE

Parts


Anderson Promoters

  • Relative Strength: weak (0.33) BBa_J23110
    • Selected as a relatively weak promoter against which to compare the expression of the PETase construct
  • Relative Strength: moderate (0.58) BBa_J23111
    • Selected as a relatively moderately strong promoter against which to compare the expression of the PETase construct
  • Relative Strength: strong (1) BBa_J23100
    • Selected as a relatively strong promoter against which to compare the expression of the PETase construct

    • Tags

  • N-OsmY
    • Fusion proteins with an N-terminal osmY have been shown to successfully secrete proteins of interest out of E. Coli
  • C-Myc
    • This C-terminal tag would allow the PETase construct to be more easily detected by Western blot assays for expression

    • Others

  • PETase sequence
    • Gene that coded for the PET hydrolase
  • Plasmid backbone
    • The pSB1C3 backbone is recommended
  • High efficiency E. Coli cells
    • Hosted the PETase constructs

    • Parts Submitted

  • Gibson-assembled pSB1C3 with novel fusion osmY-PETase A1 (BBa_K1885001)
    • This part offers efficiency in the initial step in the bioremediation of polyethylene terephthalate (PET): the production of metabolizable monomers (ethylene glycol and terephthalate) through hydrolysis. PETase catalyzes the hydrolysis of PET into terephthalate and ethylene glycol monomers. The N-terminal osmY protein and its signal peptide fused to PETase, with serine-glycine peptide’s repeating intervention, serves to secrete PETase out of the cell (this reacts with PET in the cell's local environment). The Anderson promoter, with a relative strength of 1 that precedes the fusion protein in the plasmid, serves to constitutively express the fusion PETase, enhancing the degradation of environmental PET. This part utilizes the pSB1C3 backbone and confers resistance to chloramphenicol.
  • Gibson-assembled pSB1C3 with novel fusion osmY-PETase A0.58 (BBa_K1885002)
    • This part offers efficiency in the initial step in the bioremediation of polyethylene terephthalate (PET): the production of metabolizable monomers (ethylene glycol and terephthalate) through hydrolysis. PETase catalyzes the hydrolysis of PET into terephthalate and ethylene glycol monomers. The N-terminal osmY protein and its signal peptide fused to PETase, with serine-glycine peptide’s repeating intervention, serves to secrete PETase out of the cell (this reacts with PET in the cell's local environment). The Anderson promoter, with a relative strength of 0.58 that precedes the fusion protein in the plasmid, serves to constitutively express the fusion PETase, enhancing the degradation of environmental PET. This part utilizes the pSB1C3 backbone and confers resistance to chloramphenicol.
  • Gibson-assembled pSB1C3 with novel fusion osmY-PETase A0.33 (BBa_K1885005)
    • This part offers efficiency in the initial step in the bioremediation of polyethylene terephthalate (PET): the production of metabolizable monomers (ethylene glycol and terephthalate) through hydrolysis. PETase catalyzes the hydrolysis of PET into terephthalate and ethylene glycol monomers. The N-terminal osmY protein and its signal peptide fused to PETase, with serine-glycine peptide’s repeating intervention, serves to secrete PETase out of the cell (this reacts with PET in the cell's local environment). The Anderson promoter, with a relative strength of 0.33 that precedes the fusion protein in the plasmid, serves to constitutively express the fusion PETase, enhancing the degradation of environmental PET. This part utilizes the pSB1C3 backbone and confers resistance to chloramphenicol.