Title of Document : THE TRANSCRIPTIONAL REGULATOR MPHR ( A ) : CHARACTERIZATION AND CIRCUIT ENGINEERING

Title of Document: THE TRANSCRIPTIONAL REGULATOR MPHR(A): CHARACTERIZATION AND CIRCUIT ENGINEERING Adam Smolinsky, Master of Science, 2008 Directed By: Dr. T. Ashton Cropp, Assistant Professor, Department of Chemistry and Biochemistry The MphR(A) transcriptional repressor protein was incorporated into high-efficiency expression vectors, purified and characterized. Initial screens for crystallography solvents were conducted along with preliminary gel-shift and intrinsic fluorescence quenching experiments to obtain the equilibrium dissociation constant for the protein-DNA interactions. The constant was found to be 0.5 – 1.9 M, depending on the method used. The gene was also incorporated into positive feedback circuits to detect macrolide antibiotics using various reporter genes and plasmid constructs. Qualitatively, the circuits showed a change in output upon the addition of MphR to the system. THE TRANSCRIPTIONAL REGULATOR MPHR(A): CHARACTERIZATION AND CIRCUIT ENGINEERING