RIS ID
53052
Abstract
Using 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.
Included in
Life Sciences Commons, Physical Sciences and Mathematics Commons, Social and Behavioral Sciences Commons
Publication Details
Ozawa, K., Loscha, K. V., Kuppan, K. V., Loh, C. Theng., Dixon, N. E., Otting, G. (2012). High-yield cell-free protein synthesis for site-specific incorporation of unnatural amino acids at two sites. Biochemical and Biophysical Research Communications, 418 (4), 652-656.