Difference between revisions of "Team:Waterloo/Meet the Microbes"

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<li class=""><a class="dropdown-item" href="https://2018.igem.org/Team:Waterloo/Engagement"><span>Engagement</span></a></li>
 
<li class=""><a class="dropdown-item" href="https://2018.igem.org/Team:Waterloo/Engagement"><span>Engagement</span></a></li>
 
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<li class=""><a class="dropdown-item" href="https://2018.igem.org/Team:Waterloo/Communication"><span>Communication</span></a></li>
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<li class=""><a class="dropdown-item" href="https://2018.igem.org/Team:Waterloo/IAT"><span>Iat</span></a></li>
 
</li>
 
</li>
 
<li class=""><a class="dropdown-item" href="https://2018.igem.org/Team:Waterloo/Societal_Considerations"><span>Societal Considerations</span></a></li>
 
<li class=""><a class="dropdown-item" href="https://2018.igem.org/Team:Waterloo/Societal_Considerations"><span>Societal Considerations</span></a></li>
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<div class="content">
 
 
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<div class="row"><div class="col"><div class="content-main"><h2 id="empty-jt2">Empty JT2</h2>
 
<div class="row"><div class="col"><div class="content-main"><h2 id="empty-jt2">Empty JT2</h2>
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<p><img src="https://static.igem.org/mediawiki/2018/5/56/T--Waterloo--MeetMicro_JT2.png" alt="Empty JT2"></p>
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<p><img align="left" src="https://static.igem.org/mediawiki/2018/5/56/T--Waterloo--MeetMicro_JT2.png"></p>
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<h4 id="strain">Strain</h4>
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<p><strong>Strain</strong>: <em>Escherichia coli</em> <a href="https://www.addgene.org/80403/">JT2 SKA974</a>. This strain has been engineered for optogenetic control using CcaS/R. Its genomic MetE gene is under control of the CcaS/R promoter. It also has a Kanamycin resistance gene in its genome. </p>
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<ul>
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<p><strong>Plasmids</strong>: None</p>
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<li><p><em>Escherichia coli</em> <a href="https://www.addgene.org/80403/">JT2 SKA974</a> </p>
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<p><strong>Antibiotic Resistance(s)</strong>: Kanamycin</p>
−
</li>
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<p><strong>Fluorescent Marker</strong>: None</p>
−
<li><p>This strain has been engineered for optogenetic control using CcaS/R. Its genomic MetE gene is under control of the CcaS/R promoter. It also has a Kanamycin resistance gene in its genome. </p>
+
<p><strong>Can produce methionine?</strong> No </p>
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</li>
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<hr>
−
</ul>
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−
<h4 id="plasmids">Plasmids</h4>
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−
<ul>
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−
<li>None</li>
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−
</ul>
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−
<h4 id="antibiotic-resistance-s-">Antibiotic Resistance(s)</h4>
+
−
<ul>
+
−
<li>Kanamycin</li>
+
−
</ul>
+
−
<h4 id="fluorescent-marker">Fluorescent Marker</h4>
+
−
<ul>
+
−
<li>None</li>
+
−
</ul>
+
−
<h4 id="can-produce-methionine-">Can produce methionine?</h4>
+
−
<ul>
+
−
<li>No </li>
+
−
</ul>
+
−
<h4 id="which-experiments-was-i-involved-in-">Which experiments was I involved in?</h4>
+
−
<ul>
+
−
<li>Is methionine shared between cells? </li>
+
−
<li>Biobrick <a href="http://parts.igem.org/Part:BBa_K2573000">BBa_K2573000 characterization</a> </li>
+
−
</ul>
+
 
<h2 id="jt2-ccas-r">JT2 - CcaS/R</h2>
 
<h2 id="jt2-ccas-r">JT2 - CcaS/R</h2>
−
<p><img src="https://static.igem.org/mediawiki/2018/a/a9/T--Waterloo--MeetMicro_JT2CcaSR.png" alt="JT2 CcaS/R"> </p>
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<p><img align="left" src="https://static.igem.org/mediawiki/2018/a/a9/T--Waterloo--MeetMicro_JT2CcaSR.png"></p>
−
<h4 id="strain">Strain</h4>
+
<p><strong>Strain</strong>: <em>Escherichia coli</em> <a href="https://www.addgene.org/80403/">JT2 SKA974</a>. This strain has been engineered for optogenetic control using CcaS/R. Its genomic MetE gene is under control of the CcaS/R promoter. It also has a Kanamycin resistance gene in its genome. </p>
−
<ul>
+
<p><strong>Plasmids</strong>:</p>
−
<li><em>Escherichia coli</em> <a href="https://www.addgene.org/80403/">JT2 SKA974</a>  
+
−
This strain has been engineered for optogenetic control using CcaS/R. Its genomic MetE gene is under control of the CcaS/R promoter. It also has a Kanamycin resistance gene in its genome. </li>
+
−
</ul>
+
−
<h4 id="plasmids">Plasmids</h4>
+
 
<ul>
 
<ul>
 
<li><a href="https://www.addgene.org/63197/">pSR43.6r</a> (CcaS, its photoreceptor, p15A origin of replication)</li>
 
<li><a href="https://www.addgene.org/63197/">pSR43.6r</a> (CcaS, its photoreceptor, p15A origin of replication)</li>
 
<li><a href="https://www.addgene.org/63176/">pSR58.6</a> (CcaR, sfGFP under the CcaR promoter, ColeI origin of replication)<ul>
 
<li><a href="https://www.addgene.org/63176/">pSR58.6</a> (CcaR, sfGFP under the CcaR promoter, ColeI origin of replication)<ul>
−
<li>Note that we made 2 versions of this strain. One contains both these plasmids, unaltered, and was used for characterization experiments. We cut GFP out of pSR58.6 for the second version of this strain, and used it for non-characterization experiments.  </li>
+
<li>Note that we made 2 versions of this strain. One contains both these plasmids, unaltered, and was used for characterization  
 +
experiments. We cut GFP out of pSR58.6 for the second version of this strain, and used it for non-characterization experiments.  </li>
 
</ul>
 
</ul>
 
</li>
 
</li>
 
</ul>
 
</ul>
−
<h4 id="antibiotic-resistance-s-">Antibiotic Resistance(s)</h4>
+
<p><strong>Antibiotic Resistance(s)</strong>: Kanamycin, Spectinomycin, Chloramphenicol </p>
−
<ul>
+
<p><strong>Fluorescent Marker</strong>: GFP in the characterization version of this strain, only expressed when CcaS/R is activated.</p>
−
<li>Kanamycin</li>
+
<p><strong>Can produce methionine?</strong> Yes, but only when CcaS/R is activated. </p>
−
<li>Spectinomycin</li>
+
<hr>
−
<li>Chloramphenicol </li>
+
−
</ul>
+
−
<h4 id="fluorescent-marker">Fluorescent Marker</h4>
+
−
<ul>
+
−
<li>GFP in the characterization version of this strain, only expressed when CcaS/R is activated.</li>
+
−
</ul>
+
−
<h4 id="can-produce-methionine-">Can produce methionine?</h4>
+
−
<ul>
+
−
<li>Yes, but only when CcaS/R is activated. </li>
+
−
</ul>
+
−
<h4 id="which-experiments-was-i-involved-in-">Which experiments was I involved in?</h4>
+
−
<ul>
+
−
<li>Is methionine shared between cells? </li>
+
−
<li>What is the metabolic load of (over)expressing MetE?</li>
+
−
<li>Can we control growth with red and green light? </li>
+
−
</ul>
+
 
<h2 id="empty-dh5alpha">Empty DH5alpha</h2>
 
<h2 id="empty-dh5alpha">Empty DH5alpha</h2>
−
<p><img src="https://static.igem.org/mediawiki/2018/9/92/T--Waterloo--MeetMicro_dh5.png" alt="Empty DH5"> </p>
+
<p><img align="left" src="https://static.igem.org/mediawiki/2018/9/92/T--Waterloo--MeetMicro_dh5.png"></p>
−
<h4 id="strain">Strain</h4>
+
<p><strong>Strain</strong>: <em>Escherichia coli</em> DH5alpha</p>
−
<ul>
+
<p><strong>Plasmids</strong>: None</p>
−
<li><em>Escherichia coli</em> DH5alpha </li>
+
<p><strong>Antibiotic Resistance(s)</strong> None</p>
−
</ul>
+
<p><strong>Fluorescent Marker</strong> None</p>
−
<h4 id="plasmids">Plasmids</h4>
+
<p><strong>Can produce methionine?</strong> Yes, with wild type genomic MetE gene. </p>
−
<ul>
+
<hr>
−
<li>None</li>
+
−
</ul>
+
−
<h4 id="antibiotic-resistance-s-">Antibiotic Resistance(s)</h4>
+
−
<ul>
+
−
<li>None</li>
+
−
</ul>
+
−
<h4 id="fluorescent-marker">Fluorescent Marker</h4>
+
−
<ul>
+
−
<li>None</li>
+
−
</ul>
+
−
<h4 id="can-produce-methionine-">Can produce methionine?</h4>
+
−
<ul>
+
−
<li>Yes, with my wild type genomic MetE gene. </li>
+
−
</ul>
+
−
<h3 id="which-experiments-was-i-involved-in-">Which experiments was I involved in?</h3>
+
−
<ul>
+
−
<li>Can we reliably measure 2 populations in co-culture?</li>
+
−
</ul>
+
 
<h2 id="dh5alpha-gfp">DH5alpha - GFP</h2>
 
<h2 id="dh5alpha-gfp">DH5alpha - GFP</h2>
−
<p><img src="https://static.igem.org/mediawiki/parts/9/99/T--Waterloo--DH5alpha-GFP.png" alt="DH5 GFP"></p>
+
<p><img align="left" src="https://static.igem.org/mediawiki/2018/c/c9/T--Waterloo--MeetMicro_GFP.png"></p>
−
<h3 id="strain">Strain</h3>
+
<p><strong>Strain</strong>: <em>Escherichia coli</em> DH5alpha </p>
−
<ul>
+
<p><strong>Plasmids</strong>:</p>
−
<li><em>Escherichia coli</em> DH5alpha</li>
+
−
</ul>
+
−
<h3 id="plasmids">Plasmids</h3>
+
 
<ul>
 
<ul>
 
<li><a href="http://parts.igem.org/Part:BBa_E0040">BBa_E0040 in pSB1C3</a> (GFP biobrick, ColeI origin of replication)</li>
 
<li><a href="http://parts.igem.org/Part:BBa_E0040">BBa_E0040 in pSB1C3</a> (GFP biobrick, ColeI origin of replication)</li>
 
<li>a plasmid we constructed that has Spectinomycin resistance and p15A origin of replication</li>
 
<li>a plasmid we constructed that has Spectinomycin resistance and p15A origin of replication</li>
 
</ul>
 
</ul>
−
<h3 id="antibiotic-resistance-s-">Antibiotic Resistance(s)</h3>
+
<p><strong>Antibiotic Resistance(s)</strong>: Spectinomycin, Chloramphenicol </p>
−
<ul>
+
<p><strong>Fluorescent Marker</strong>: GFP, constitutively expressed. </p>
−
<li>Spectinomycin</li>
+
<p><strong>Can produce methionine?</strong>: Yes, with wild type genomic MetE gene. </p>
−
<li>Chloramphenicol </li>
+
−
</ul>
+
−
<h3 id="fluorescent-marker">Fluorescent Marker</h3>
+
−
<ul>
+
−
<li>GFP, constitutively expressed. </li>
+
−
</ul>
+
−
<h3 id="can-produce-methionine-">Can produce methionine?</h3>
+
−
<ul>
+
−
<li>Yes, with my wild type genomic MetE gene. </li>
+
−
</ul>
+
−
<h3 id="which-experiments-was-i-involved-in-">Which experiments was I involved in?</h3>
+
−
<ul>
+
−
<li>Can we reliably measure 2 populations in co-culture?</li>
+
−
</ul>
+
 
</div></div></div>
 
</div></div></div>
 
</div>
 
</div>
 
</html>
 
</html>
 
{{Waterloo/footer}}
 
{{Waterloo/footer}}

Revision as of 01:35, 18 October 2018

Meet the Microbes

Empty JT2

Strain: Escherichia coli JT2 SKA974. This strain has been engineered for optogenetic control using CcaS/R. Its genomic MetE gene is under control of the CcaS/R promoter. It also has a Kanamycin resistance gene in its genome.

Plasmids: None

Antibiotic Resistance(s): Kanamycin

Fluorescent Marker: None

Can produce methionine? No

JT2 - CcaS/R

Strain: Escherichia coli JT2 SKA974. This strain has been engineered for optogenetic control using CcaS/R. Its genomic MetE gene is under control of the CcaS/R promoter. It also has a Kanamycin resistance gene in its genome.

Plasmids:

  • pSR43.6r (CcaS, its photoreceptor, p15A origin of replication)
  • pSR58.6 (CcaR, sfGFP under the CcaR promoter, ColeI origin of replication)
    • Note that we made 2 versions of this strain. One contains both these plasmids, unaltered, and was used for characterization experiments. We cut GFP out of pSR58.6 for the second version of this strain, and used it for non-characterization experiments.

Antibiotic Resistance(s): Kanamycin, Spectinomycin, Chloramphenicol

Fluorescent Marker: GFP in the characterization version of this strain, only expressed when CcaS/R is activated.

Can produce methionine? Yes, but only when CcaS/R is activated.

Empty DH5alpha

Strain: Escherichia coli DH5alpha

Plasmids: None

Antibiotic Resistance(s) None

Fluorescent Marker None

Can produce methionine? Yes, with wild type genomic MetE gene.

DH5alpha - GFP

Strain: Escherichia coli DH5alpha

Plasmids:

  • BBa_E0040 in pSB1C3 (GFP biobrick, ColeI origin of replication)
  • a plasmid we constructed that has Spectinomycin resistance and p15A origin of replication

Antibiotic Resistance(s): Spectinomycin, Chloramphenicol

Fluorescent Marker: GFP, constitutively expressed.

Can produce methionine?: Yes, with wild type genomic MetE gene.

Waterloo iGEM 2018

© 2018 Waterloo iGEM