WITH ABIDEEN AYANGBEMI, CHRIS HATCHELL, AND DR. DAVID JACOBSON, DEPARTMENT OF CHEMISTRY
Sfp Function for Atomic Force Microscopy
Expressed in the BL21 DE3 strain of E. coli via plasmid transformation, the Sfp protein can be used in attachment chemistry as a link between molecules. Single molecule-experiments help explain the folding and dynamics of biomolecules. Sfp can be used in single molecule pulling experiments because it catalyzes a covalent bond between the ybbr amino acid tag and Coenzyme A (CoA). In past experiments in Dr. Jacobson’s lab, this expression has yielded Sfp protein, but it has not been functional. The goal of this current trial is to produce a new sample of functional Sfp to enable this attachment chemistry. A gel shift assay will be used to monitor the binding of Sfp to a test polymer. Nucleic acids (RNA and DNA) are some of the most essential organic molecules of biology and can be probed in the single molecule pulling experiments. Both molecules can have different sugar structures, but which ones they exist in are still in question. With functional Sfp and Atomic Force Microscopy the thermodynamic stability discrepancies of the compact and extended structures of the RNA sugar pucker can be tested and analyzed for a better understanding of these crucial molecules.
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