Cell-free translation reconstituted with purified components

Authors
Shimizu, Y Inoue, A Tomari, Y Suzuki, T Yokogawa, T Nishikawa, K Ueda, T
Citation
Y. Shimizu et al., Cell-free translation reconstituted with purified components, NAT BIOTECH, 19(8), 2001, pp. 751-755
Citations number
26
Categorie Soggetti
Biotecnology & Applied Microbiology",Microbiology
Journal title
NATURE BIOTECHNOLOGY
ISSN journal
10870156 → ACNP
Volume
19
Issue
8
Year of publication
2001
Pages
751 - 755
Database
ISI
SICI code
1087-0156(200108)19:8<751:CTRWPC>2.0.ZU;2-S
Abstract
We have developed a protein-synthesizing system reconstituted from recombin ant tagged protein factors purified to homogeneity. The system was able to produce protein at a rate of about 160 mug/ml/h in a batch mode without the need for any supplementary apparatus. The protein products were easily pur ified within 1 h using affinity chromatography to remove the tagged protein factors. Moreover, omission of a release factor allowed efficient incorpor ation of an unnatural amino acid using suppressor transfer RNA (tRNA).