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

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<div class="row"><div class="col"><div class="content-main"></div></div></div>
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<div class="row"><div class="col"><div class="content-main"><h2 id="empty-jt2">Empty JT2</h2>
 +
<p><img src="https://static.igem.org/mediawiki/2018/5/56/T--Waterloo--MeetMicro_JT2.png" alt="Empty JT2"></p>
 +
<h4 id="strain">Strain</h4>
 +
<ul>
 +
<li><p><em>Escherichia coli</em> <a href="https://www.addgene.org/80403/">JT2 SKA974</a> </p>
 +
</li>
 +
<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>
 +
</li>
 +
</ul>
 +
<h4 id="plasmids">Plasmids</h4>
 +
<ul>
 +
<li>None</li>
 +
</ul>
 +
<h4 id="antibiotic-resistance-s-">Antibiotic Resistance(s)</h4>
 +
<ul>
 +
<li>Kanamycin</li>
 +
</ul>
 +
<h4 id="growth-rate">Growth Rate</h4>
 +
<ul>
 +
<li><h4 id="fluorescent-marker">Fluorescent Marker</h4>
 +
</li>
 +
<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>
 +
<p><img src="https://static.igem.org/mediawiki/2018/a/a9/T--Waterloo--MeetMicro_JT2CcaSR.png" alt="JT2 CcaS/R"> </p>
 +
<h4 id="strain">Strain</h4>
 +
<ul>
 +
<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>
 +
<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>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>
 +
</li>
 +
</ul>
 +
<h4 id="antibiotic-resistance-s-">Antibiotic Resistance(s)</h4>
 +
<ul>
 +
<li>Kanamycin</li>
 +
<li>Spectinomycin</li>
 +
<li>Chloramphenicol </li>
 +
</ul>
 +
<h4 id="growth-rate">Growth Rate</h4>
 +
<ul>
 +
<li><h4 id="fluorescent-marker">Fluorescent Marker</h4>
 +
</li>
 +
<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>
 +
<p><img src="https://static.igem.org/mediawiki/2018/9/92/T--Waterloo--MeetMicro_dh5.png" alt="Empty DH5"> </p>
 +
<h4 id="strain">Strain</h4>
 +
<ul>
 +
<li><em>Escherichia coli</em> DH5alpha </li>
 +
</ul>
 +
<h4 id="plasmids">Plasmids</h4>
 +
<ul>
 +
<li>None</li>
 +
</ul>
 +
<h4 id="antibiotic-resistance-s-">Antibiotic Resistance(s)</h4>
 +
<ul>
 +
<li>None</li>
 +
</ul>
 +
<h4 id="growth-rate">Growth Rate</h4>
 +
<ul>
 +
<li><h4 id="fluorescent-marker">Fluorescent Marker</h4>
 +
</li>
 +
<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>
 +
<p><img src="" alt="DH5 GFP"></p>
 +
<h3 id="strain">Strain</h3>
 +
<ul>
 +
<li><em>Escherichia coli</em> DH5alpha</li>
 +
</ul>
 +
<h3 id="plasmids">Plasmids</h3>
 +
<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 plasmid we constructed that has Spectinomycin resistance and p15A origin of replication</li>
 +
</ul>
 +
<h3 id="antibiotic-resistance-s-">Antibiotic Resistance(s)</h3>
 +
<ul>
 +
<li>Spectinomycin</li>
 +
<li>Chloramphenicol </li>
 +
</ul>
 +
<h3 id="growth-rate">Growth Rate</h3>
 +
<ul>
 +
<li><h3 id="fluorescent-marker">Fluorescent Marker</h3>
 +
</li>
 +
<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>
 
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Revision as of 23:02, 17 October 2018

Meet the Microbes

Empty JT2

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

Growth Rate

  • Fluorescent Marker

  • None

Can produce methionine?

  • No

Which experiments was I involved in?

JT2 - CcaS/R

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

Growth Rate

  • 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.

Which experiments was I involved in?

  • Is methionine shared between cells?
  • What is the metabolic load of (over)expressing MetE?
  • Can we control growth with red and green light?

Empty DH5alpha

Empty DH5

Strain

  • Escherichia coli DH5alpha

Plasmids

  • None

Antibiotic Resistance(s)

  • None

Growth Rate

  • Fluorescent Marker

  • None

Can produce methionine?

  • Yes, with my wild type genomic MetE gene.

Which experiments was I involved in?

  • Can we reliably measure 2 populations in co-culture?

DH5alpha - GFP

DH5 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

Growth Rate

  • Fluorescent Marker

  • GFP, constitutively expressed.

Can produce methionine?

  • Yes, with my wild type genomic MetE gene.

Which experiments was I involved in?

  • Can we reliably measure 2 populations in co-culture?

Waterloo iGEM 2018

© 2018 Waterloo iGEM