Team:Cambridge/Project/FluorideRiboswitch

From 2012.igem.org

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==Fluoride riboswitch==
==Fluoride riboswitch==
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We also plan on implementing and characterising a Fluoride riboswitch. This, unlike the Magnesium construct, is a positive regulator. The riboswitch, originally isolated from Bacillus cereus, serves as a transcriptional attenuator in the abscence of fluoride. In the presence of fluoride its conformation changes and the repression is lifted. In B. cereus this serves to permit translation of a fluoride efflux pump, which allows the bacteria to cope with the potentially toxic fluoride levels in which it finds itself.
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We also plan on implementing and characterising a Fluoride riboswitch. This, unlike the Magnesium construct, is a positive regulator. The riboswitch, originally isolated from Bacillus cereus, serves as a transcriptional attenuator in the absence of fluoride. In the presence of fluoride its conformation changes and the repression is lifted. In B. cereus this serves to permit translation of a fluoride efflux pump, which allows the bacteria to cope with the potentially toxic fluoride levels in which it finds itself.
We have kindly been provided with a plasmid from the [[Team:Cambridge/Special_thanks|Breaker lab]] in Yale University, the key features of which are shown below:
We have kindly been provided with a plasmid from the [[Team:Cambridge/Special_thanks|Breaker lab]] in Yale University, the key features of which are shown below:
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[[File: F-riboswitch.png|750px|center|thumb|Our riboswitch construct for the detection of fluoride.]]
[[File: F-riboswitch.png|750px|center|thumb|Our riboswitch construct for the detection of fluoride.]]
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This construct was initially tested with an X-GAL assay, which produced excellent results. Having gained this qualatative data, we moved onto performing a Miller assay to produce more quantatative data.
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For the results of our characterisation assays, please see our [[Team:Cambridge/Project/Results|<span style="color:#000066"><u>Results page</u></span>]]
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[[File:fluoridegradient.jpg|left|250px|thumb|The results of our X-GAL assay on the fluoride riboswitch construct. Numbers on the top of the tubes represent the fluoride concentration of each solution in mM.]]
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If time permits, we are also aiming to put our part into [[Team:Cambridge/Project/Standardised_Outputs|Ratiometrica]], once it is completed:
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===References===
===References===

Revision as of 02:15, 27 September 2012

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Fluoride riboswitch

We also plan on implementing and characterising a Fluoride riboswitch. This, unlike the Magnesium construct, is a positive regulator. The riboswitch, originally isolated from Bacillus cereus, serves as a transcriptional attenuator in the absence of fluoride. In the presence of fluoride its conformation changes and the repression is lifted. In B. cereus this serves to permit translation of a fluoride efflux pump, which allows the bacteria to cope with the potentially toxic fluoride levels in which it finds itself.

We have kindly been provided with a plasmid from the Breaker lab in Yale University, the key features of which are shown below:

Our riboswitch construct for the detection of fluoride.

For the results of our characterisation assays, please see our Results page

References

  • Jenny L. Baker et al., Widespread Genetic Switches and Toxicity Resistance Proteins for Fluoride, Science 335, 233-235.

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