High-yield cell-free protein synthesis for site-specific incorporation of unnatural amino acids at two sitesFaculty of Science - Papers (Archive)
AbstractUsing aminoacyl-tRNA synthetase/suppressor tRNA pairs derived from Methanocaldococcus jannaschii, an Escherichia coli cell-free protein production system affords proteins with site-specifically incorporated unnatural amino acids (UAAs) in high yields through the use of optimized amber suppressor tRNACUA opt and optimization of reagent concentrations. The efficiency of the cell-free system allows the incorporation of trifluoromethyl-phenylalanine using a polyspecific synthetase evolved previously for p-cyanophenylalanine, and the incorporation of UAAs at two different sites of the same protein without any re-engineering of the E. coli cells used to make the cell-free extract.
Citation InformationKiyoshi Ozawa, Karin V Loscha, Kekini V Kuppan, Choy Theng Loh, et al.. "High-yield cell-free protein synthesis for site-specific incorporation of unnatural amino acids at two sites" (2012) p. 652 - 656
Available at: http://works.bepress.com/nick_dixon/30/