Difference between revisions of "Team:East Chapel Hill/Demonstrate"
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Revision as of 06:42, 1 November 2017
Demonstrate
Our project pertains to measuring concentrations of fluoride and characterizing fluoride riboswitches with higher affinity to fluoride; these technologies can be used to determine methods to sequester, bioremediate, and detect fluoride.
We developed the Chloramphenicol Acetyltransferase Operon (CHOP) regulated by the fluoride riboswitch as a system to characterize the fluoride riboswitch.
Since bacteria does not grow in the presence of chloramphenicol, the protein bacteria needs chloramphenicol acetyltransferase in order to build resistance to chloramphenicol and survive.
We plated CHOP and ΔcrcB, which is our control E.coli without the resistance to chloramphenicol, onto plates with no fluoride and increased concentrations of fluoride in order to test the best level the fluoride riboswitch can grow at.
Our results demonstrated that with no fluoride there was no growth of CHOP. However, the growth of CHOP increased as the concentration of fluoride increased. In other words, CHOP is fluoride dependent. Our fluoride riboswitch was able to regulate the chloramphenicol acetyltransferase and allow the bacteria to survive; the concentration with the most growth of CHOP would represent the best activation concentration of fluoride for the riboswitch.
Since the growth of CHOP is correlated with the activation of the fluoride riboswitch, CHOP can be used to find other riboswitches that have a better affinity to fluoride by finding the lowest level of fluoride CHOP can grow at specific riboswitches.
