کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
1929889 | 1050477 | 2012 | 5 صفحه PDF | دانلود رایگان |

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 tRNACUAopt 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-cyano-phenylalanine, 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.
Cell-free synthesis.Figure optionsDownload as PowerPoint slideHighlights
► Cell-free synthesis affords high protein yields with unnatural amino acids.
► 4-(Trifluoromethyl)phenylalanine incorporated by a polyspecific synthetase.
► Truncation at amber stop codons can be suppressed almost completely.
► Two unnatural amino acids can readily be incorporated at two different sites.
Journal: Biochemical and Biophysical Research Communications - Volume 418, Issue 4, 24 February 2012, Pages 652–656