Team:Evry/Sensor

From 2013.igem.org

(Difference between revisions)
Line 9: Line 9:
<p>
<p>
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 <a href="https://2013.igem.org/Team:Evry/Project_FUR">Fur system</a> to repress a target gene.
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 <a href="https://2013.igem.org/Team:Evry/Project_FUR">Fur system</a> to repress a target gene.
 +
</p>
 +
<p>
 +
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.
</p>
</p>
<br>
<br>

Revision as of 00:50, 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, fes, fepA and yncE. Finally, pAceB appears to be the best candidate to build our sensor system.



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.