Goals with GFP with Unnatural Amino Acids

In my last post I discussed how my primary goal for the summer was to synthesize different types of GFP, all with some unnatural amino acid (UAA) inserted into the protein sequence. The UAAs that I inserted into GFP were 3-fluorotyrosine (3FY), ortho-nitrobenzyl tyrosine (ONBY), and p-propargyloxyphenylalanine (pPrF). The goal of inserting these UAAs into the protein is to observe how they modify the chemical properties of the GFP fluorophore. When analyzed with a spectrophotometer, wild-type GFP has a characteristic spectrum. Because the UAAs are inserted in the fluorophore region of the peptide, a shift in the fluorescent spectrum is apparent in GFP with UAAs incorporated. The next step in this project was analyze the protein itself using spectroscopy and conducting experiments on the protein to see if there is a shift in the fluorescence spectra.

Thanks for reading!