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Nat Biotechnol. 2001 Aug;19(8):751-5.

Cell-free translation reconstituted with purified components.

Author information

1
Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, Bldg FSB401, 5-1-5, Kasiwanoha, Kashiwa, Chiba prefecture 277-8562, Japan.

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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PMID:
11479568
DOI:
10.1038/90802
[Indexed for MEDLINE]

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