Team:Evry/Sensor
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- | We constructed iron sensors 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. | + | We constructed iron sensors 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 using the <a href="https://2013.igem.org/Team:Evry/Project_FUR">Fur system. |
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Revision as of 10:28, 4 October 2013
Iron Sensor
We constructed iron sensors 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 using the Fur system.
Fig 1 Diagram of a 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.
Table I Genetic elements used to make iron-responsive sensors.
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 |