Nature Biotechnology
19, 751 - 755 (2001)
doi:10.1038/90802
Cell-free translation reconstituted with purified componentsYoshihiro Shimizu1, Akio Inoue2, Yukihide Tomari1, Tsutomu Suzuki2, Takashi Yokogawa3, Kazuya Nishikawa3
& Takuya Ueda21
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. 2
Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Bldg FSB401, 5-1-5, Kasiwanoha, Kashiwa, Chiba prefecture 277-8562, Japan. 3
Department of Biomolecular Science, Faculty of Engineering, Gifu University, Yanagido, Gifu 501-1193, Japan.
Correspondence should be addressed to Takuya Ueda ueda@k.u-tokyo.ac.jpWe 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  g/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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