Presenter(s): Michael Veirs
Faculty Mentor(s): Julia Widom
Poster 23
Session: Sciences
Fluorescence Resonance Energy Transfer (FRET) is a photophysical phenomenon in which the excitation of a donor fluorophore results in fluorescent emission from an acceptor fluorophore when the two molecules are in close proximity. Using a FRET system, we intend to investigate the folding dynamics of the preQ1 riboswitch, which is an RNA species that regulates gene expression in bacteria. We used a double-stranded DNA system and the fluorescent adenine analog 2-aminopurine (2AP) to determine fluorescent molecules that can be used as FRET acceptors for 2AP. We found mFluor violet 450 and Atto390 to be appropriate acceptor fluorophores for use in more complex RNA systems. We also found that the riboswitch has a tendency to dimerize under our experimental conditions. To investigate this process, we ran our RNA samples using polyacrylamide gel electrophoresis. We determined protocols for minimizing dimerization of our RNA by varying the procedure by which the construct was annealed and stored. These results lay the foundation for using FRET to study the folding of this riboswitch and, by extension, the mechanism by which it regulates gene expression.