Difference between revisions of "Team:East Chapel Hill"

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<p style="padding-bottom:0; padding-right: 10%; padding-left:10%; black; font-size:14px;" class="big"> In many poor countries, excess concentrations of fluoride have entered water sources through erosion of sediment and minerals. These toxic concentrations, which are defined by the World Health Organization to be above 1.0mg/L, may result in an array of health complications. Fluoride has been known to induce cell stress, which in turn impairs the function of ameloblasts who are tasked with forming dental enamel. The resulting disease is dental fluorosis, which manifests in the teeth as porous and yellowed enamel. Other potential consequences of exposure to toxic fluoride levels include skeletal fluorosis, which may result in bone deformities, as well as impaired development.
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<p style="padding-bottom:0; padding-right: 10%; padding-left:10%; black; font-size:14px;" class="big"> Fluoride, in appropriate quantities, has been recognized as beneficial for protecting tooth enamel from decay. However, a significant problem arises when excess amounts of fluoride infiltrate drinking water. These high concentrations result in dental fluorosis, which is characterized in children by hypomineralization of the enamel. To address this, we are using the previously characterized fluoride riboswitch. Last year we developed an operon that, when fluoride binds, activates the riboswitch resulting in the transcription of the chloramphenicol acetyltransferase gene. Thus, when fluoride is present, bacterial growth can be observed in the presence of chloramphenicol. So that the fluoride riboswitch may detect lower concentrations of fluoride, we used restriction enzymes to test several promoters. We tested two different sequences of the fluoride riboswitch, labeled FRS1 and FRS2, to examine how their predicted folding pattern would change the efficacy of the operon. We found the OXB18 promoter and FRS1 to be most successful in promoting bacterial growth
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Our project involves the development of an operon that will serve as a visual indicator of excess fluoride. We plan to further develop and improve the operon created by last year’s iGEM team, specifically focusing on the fluoride riboswitch and promoter. We are looking for promoters that have an increased gene expression and riboswitches that have increased affinity to fluoride. This would allow for our operon to detect fluoride at levels even lower than 1.0mg/L, a significant improvement from the previous operon.
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Revision as of 02:58, 14 September 2018

East Chapel Hill
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Improving the Efficacy of the Fluoride Riboswitch as a Visual Detector of Fluoride Concentrations in Water

Fluoride, in appropriate quantities, has been recognized as beneficial for protecting tooth enamel from decay. However, a significant problem arises when excess amounts of fluoride infiltrate drinking water. These high concentrations result in dental fluorosis, which is characterized in children by hypomineralization of the enamel. To address this, we are using the previously characterized fluoride riboswitch. Last year we developed an operon that, when fluoride binds, activates the riboswitch resulting in the transcription of the chloramphenicol acetyltransferase gene. Thus, when fluoride is present, bacterial growth can be observed in the presence of chloramphenicol. So that the fluoride riboswitch may detect lower concentrations of fluoride, we used restriction enzymes to test several promoters. We tested two different sequences of the fluoride riboswitch, labeled FRS1 and FRS2, to examine how their predicted folding pattern would change the efficacy of the operon. We found the OXB18 promoter and FRS1 to be most successful in promoting bacterial growth