Abstract
The incorporation of unnatural amino acids (UAAs) into bacterial proteins provide insight for understanding protein interactions and have implications within the medical field. These small UAA molecules, not endogenous to the biological systems, have unique properties and can be incorporated into proteins. Traditionally, TAG is a codon within DNA that signals the termination of polypeptide formation. Incorporating UAAs requires the use of a mutated orthogonal tRNA/synthetase pair to encode the unnatural amino acid, the amber stop codon (TAG) is incorporated through site-directed mutagenesis to the DNA sequence. Here, a fluorescent protein will be modified to incorporate the UAA, p-boronophenylalanine, resulting in a change in fluorescence that may be monitored upon UAA incorporation. Further implications to this type of work include the usage of subsequent reactions which consist of the addition of diols via linking chemistry.