کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1929889 1050477 2012 5 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
High-yield cell-free protein synthesis for site-specific incorporation of unnatural amino acids at two sites
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
High-yield cell-free protein synthesis for site-specific incorporation of unnatural amino acids at two sites
چکیده انگلیسی

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.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochemical and Biophysical Research Communications - Volume 418, Issue 4, 24 February 2012, Pages 652–656
نویسندگان
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