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High-yield cell-free protein synthesis for site-specific incorporation of unnatural amino acids at two sites

Ozawa, Kiyoshi; Loscha, Karin; Kuppan, Kekini V; Loh, Choy; Dixon, Nicholas Edward; Otting, Gottfried

Description

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...[Show more]

CollectionsANU Research Publications
Date published: 2012
Type: Journal article
URI: http://hdl.handle.net/1885/60845
Source: Biochemical and Biophysical Research Communications
DOI: 10.1016/j.bbrc.2012.01.069

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