Team:Cornell/testing/project/wetlab/4/7

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<div class="row last-ele"><div class="twelve columns"><h3>Registry of Standard Parts Submitted</h3></div></div>
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<h2 class="centered">Reactor Testing</h2>
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<tr><th style="width:8px" class="header"></th><th style="width:8px" class="header"></th><th style="width:8px" class="header"></th><th style="width: 87px;" class="header">Name</th><th style="width: 80px" colspan="" class="header">Type</th><th style="width: auto;" colspan="" class="header">Description</th><th style="width: 150px" colspan="" class="header">Designer</th><th style="width: 50px" colspan="" class="header">Length</th>
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<h3>Overview of Characterization in Bioelectrochemical Stystems</h3>
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<td class="status_cell heart13">&nbsp;</td><td class="status_cell cell_white">&nbsp;</td><td class="status_cell cell_white">&nbsp;&nbsp;</td><td><a class="noul_link" href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K957000">BBa_K957000</a></td><td>Generator</td><td>Arsenic inducible mtrB with cut sites flanking RBS</td><td width="100px">Dylan Patrick Webster</td><td align="right">2625</td>
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As described in the <a href="https://2012.igem.org/Team:Cornell/testing/project/wetlab/2">Chassis</a> section, <i>S. oneidensis</i> MR-1 is capable of shuttling electrons through the Mtr pathway to reduce Fe(III) to Fe(II) because of the negative free energy change associated with the redox reaction. Thus, to encourage <i>Shewanella</i> to transfer electrons to an electrode, we poise the redox potential of an electrode to that of Fe(III)/Fe(II) in a three electrode system, controlled by a potentiostat. [[LINK TO THE PAGE ON THE POTENTIOSTAT]]. In short, this instrument works by setting the potential of a working electrode (WE) with respect to a Ag/AgCl reference eletrode (RE) by injecting current through a counter electrode (CE). These electrodes can be seen in the schematic representation of our single-compartment bioelectrochemical reactors shown below.
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<td class="status_cell heart13">&nbsp;</td><td class="status_cell cell_white">&nbsp;</td><td class="status_cell cell_white">&nbsp;&nbsp;</td><td><a class="noul_link" href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K957001">BBa_K957001</a></td><td>Generator</td><td>Arsenic inducible mtrB</td><td width="100px">Dylan Patrick Webster</td><td align="right">2704</td>
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<td class="status_cell heart13">&nbsp;</td><td class="status_cell cell_white">&nbsp;</td><td class="status_cell cell_white">&nbsp;&nbsp;</td><td><a class="noul_link" href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K957002">BBa_K957002</a></td><td>Generator</td><td>Salicylate inducible mtrB with cut sites flanking RBS </td><td width="100px">Dylan Patrick Webster</td><td align="right">3207</td>
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<h3>First Lessons Learned: Control Reactors</h3>
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<td class="status_cell heart13">&nbsp;</td><td class="status_cell cell_white">&nbsp;</td><td class="status_cell cell_white">&nbsp;&nbsp;</td><td><a class="noul_link" href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K957003">BBa_K957003</a></td><td>Generator</td><td>Salicylate inducible mtrB</td><td width="100px">Dylan Patrick Webster</td><td align="right">3284</td>
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In order to enhance the field-deployability of our final device, we initially decided to feed our reactors with LB, since a very concentrated LB source fed at a low flow rate could sustain our field reactors for extended periods of time without taking up much physical space. However, upon setting up control reactors&#8212;both in batch and continuous flow operation&#8212;we discovered that wild type <i>S. oneidensis</i> MR-1 produced significantly less current when fed with LB than M4&#8212;a commonly used media for <i>Shewanella</i>-inoculated bioelectrochemical systems.
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<td class="status_cell heart13">&nbsp;</td><td class="status_cell cell_white">&nbsp;</td><td class="status_cell cell_white">&nbsp;&nbsp;</td><td><a class="noul_link" href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K957004">BBa_K957004</a></td><td>Coding</td><td>nah Operon for degradation of naphthalene to salicylate</td><td width="100px">Dylan Patrick Webster</td><td align="right">9409</td>
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<h3>Naphthalene & Salicylate Sensing</h3>
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<td class="status_cell heart13">&nbsp;</td><td class="status_cell cell_white">&nbsp;</td><td class="status_cell cell_white">&nbsp;&nbsp;</td><td><a class="noul_link" href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K957005">BBa_K957005</a></td><td>Generator</td><td> Constitutive expression of nah Operon for degradation of naphthalene to salicylate </td><td width="100px">Dylan Patrick Webster</td><td align="right">9452</td>
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First characterized naphthalene sensors with salicylate, since our system indirectly senses naphthalene via salicylate.
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Uninduced salicylate reporter produces same current as MR-1 in rich M4 media.
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<td class="status_cell  "></td><td class="status_cell cell_white">&nbsp;</td><td class="status_cell cell_white">&nbsp;</td><td><a class="noul_link" href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K957006">BBa_K957006</a></td><td>Composite</td><td>Constitutive expression of nah Operon with oriT </td><td width="100px">Dylan Patrick Webster</td><td align="right">9831</td>
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First attempt to diagnose problem: Switch to minimal M4... Producing higher current, but we're still not getting a salicylate response.
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Next attempt: Poise the working electrode at a higher potential in attempt to get more MtrC and MtrA for MtrB to associate with. Didn't quite do the trick either.
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<h3>Arsenic Sensing</h3>
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Latest revision as of 03:48, 4 October 2012

Registry of Standard Parts Submitted

NameTypeDescriptionDesignerLength
    BBa_K957000GeneratorArsenic inducible mtrB with cut sites flanking RBSDylan Patrick Webster2625
    BBa_K957001GeneratorArsenic inducible mtrBDylan Patrick Webster2704
    BBa_K957002GeneratorSalicylate inducible mtrB with cut sites flanking RBS Dylan Patrick Webster3207
    BBa_K957003GeneratorSalicylate inducible mtrBDylan Patrick Webster3284
    BBa_K957004Codingnah Operon for degradation of naphthalene to salicylateDylan Patrick Webster9409
    BBa_K957005Generator Constitutive expression of nah Operon for degradation of naphthalene to salicylate Dylan Patrick Webster9452
  BBa_K957006CompositeConstitutive expression of nah Operon with oriT Dylan Patrick Webster9831

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