Team:Evry/Sensor

From 2013.igem.org

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We constructed 4 differents iron sensor using promoters regions from <i>aceB</i>(<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1163102">BBa_K1163102</a>), <i>fes</i>(<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1163108">BBa_K1163108</a>), <i>fepA</i>(<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1163105">BBa_K1163105</a>) and  <i>yncE</i>(<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1163111">BBa_K1163111</a>). Finally, pAceB appears to be the best candidate to build our sensor system. <a href="https://2013.igem.org/Team:Evry/Results">See our results</a>
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We constructed 4 differents iron sensor using promoters regions from <i>aceB</i> (<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1163102">BBa_K1163102</a>), <i>fes</i> (<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1163108">BBa_K1163108</a>), <i>fepA</i> (<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1163105">BBa_K1163105</a>) and  <i>yncE</i> (<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1163111">BBa_K1163111</a>). Finally, pAceB appears to be the best candidate to build our sensor system. <a href="https://2013.igem.org/Team:Evry/Results">See our results</a>
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Revision as of 01:03, 5 October 2013

Iron coli project

Iron Sensor

We constructed iron-responsive biosensors by combining 3 genetic parts: an E. coli promoter with a Ferric Uptake Regulator (Fur) binding site, a fluorescent reporter (sfGFP), and a transcriptional terminator. These sensors respond to ambient iron by using the Fur system to repress a target gene.

We constructed 4 differents iron sensor using promoters regions from aceB (BBa_K1163102), fes (BBa_K1163108), fepA (BBa_K1163105) and yncE (BBa_K1163111). Finally, pAceB appears to be the best candidate to build our sensor system. See our results



Fig 1 Diagram of our genetic iron sensor. Iron binds the Ferric Uptake Regulator (Fur) to form a complex with high affinity for the Fur box in the promoter, here shown as the aceB promoter. Once the iron-Fur complex is bound to the promoter, it represses transcription of the target gene GFP. GFP expression is thus negatively correlated with iron availability.



Fig 2 Construction of an iron-responsive genetic element by fusing a Fur-regulated promoter with a reporter gene. Promoter-reporter fusions were made with flanking restriction sites that are compatible with Biobrick-based cloning.

NAME FIGURE DESCRIPTION

E. coli promoter with Fur binding site

iron-Fur complex binds promoter to repress expression

sfGFP

Fluorescent reporter gene

Terminator

terminator to stop transcription

Plasmid

Biobrick-compatible plasmid backbone

Table I Genetic elements used to make iron-responsive sensors.