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Team:Cambridge/Project/FluorideRiboswitch
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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. | 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 [[Team:Cambridge/Special_thanks|Breaker lab]] in Yale University, the key features of which are shown below: | |
[[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.]] | ||
| + | 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 [[Team:Cambridge/Project/Standardised_Outputs|Ratiometrica]], once it is completed: | ||
===References=== | ===References=== | ||
Revision as of 20:10, 25 September 2012
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:
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.
