Team:Cambridge/Project/FluorideRiboswitch

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Previous iGEM teams have charaterised an impressive array of inducible promoters, along with other elements of biosensing circuitry... Read More






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 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.

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.

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.

If time permits, we are also aiming to put our part into Ratiometrica, once it is completed:

References

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

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