Team:Evry/Sensor
From 2013.igem.org
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- | We constructed 4 differents iron sensor using promoters regions from <i>aceB</i>, <i>fes</i>, <i>fepA</i> and <i>yncE</i>. Finally, pAceB appears to be the best candidate to build our sensor system. | + | 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 01:02, 5 October 2013
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.