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      Cell-free translation reconstituted with purified components.

      Nature biotechnology
      Amino Acids, chemistry, Cell-Free System, Chromatography, Affinity, Electrophoresis, Polyacrylamide Gel, Escherichia coli, metabolism, Histidine, Plasmids, Protein Biosynthesis, RNA, Transfer, RNA, Transfer, Amino Acyl, Recombinant Proteins, Ribosomes, Time Factors

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          Abstract

          We have developed a protein-synthesizing system reconstituted from recombinant tagged protein factors purified to homogeneity. The system was able to produce protein at a rate of about 160 microg/ml/h in a batch mode without the need for any supplementary apparatus. The protein products were easily purified within 1 h using affinity chromatography to remove the tagged protein factors. Moreover, omission of a release factor allowed efficient incorporation of an unnatural amino acid using suppressor transfer RNA (tRNA).

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