| 초록 |
The ability to incorporate unnatural amino acids can expand the scope of protein engineering beyond the present limitations of using 20 canonical amino acids. While cell-based production of unnatural amino acid-containing protein is cumbersome and time and labor-intensive process, herein we propose a rapid and efficient tool for translational incorporation of unnatural amino acids into proteins. The advantages of cell-free protein synthesis include speed and throughput in protein production. In addition, the open nature of cell-free protein synthesis allows direct modifications of the components for protein synthesis. While the simple replacements of amino acids led to mixed synthesis of proteins containing unnatural and natural amino acids, in our experiments to replace tyrosine with 3,4-dihydroxy-L-phenylalanine (L-DOPA), almost exclusive incorporation of unnatural amino acids was achieved by washing the cell extract to remove residual natural amino acids in cell extract. Proposed strategy should be able to provide a facile route to selective incorporation unnatural amino acids in protein structure. |
