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High-yield cell-free protein synthesis for site-specific incorporation of unnatural amino acids at two sites
Faculty of Science - Papers (Archive)
  • Kiyoshi Ozawa, University of Wollongong
  • Karin V Loscha, Australian National University
  • Kekini V Kuppan, Australian National University
  • Choy Theng Loh, Australian National University
  • Nicholas E Dixon, University of Wollongong
  • Gottfried Otting, Australian National University
RIS ID
53052
Publication Date
1-1-2012
Publication Details

Ozawa, K., Loscha, K. V., Kuppan, K. V., Loh, C. Theng., Dixon, N. E., Otting, G. (2012). High-yield cell-free protein synthesis for site-specific incorporation of unnatural amino acids at two sites. Biochemical and Biophysical Research Communications, 418 (4), 652-656.

Abstract

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 tRNACUA opt and optimization of reagent concentrations. The efficiency of the cell-free system allows the incorporation of trifluoromethyl-phenylalanine using a polyspecific synthetase evolved previously for p-cyanophenylalanine, and the incorporation of UAAs at two different sites of the same protein without any re-engineering of the E. coli cells used to make the cell-free extract.

Citation Information
Kiyoshi Ozawa, Karin V Loscha, Kekini V Kuppan, Choy Theng Loh, et al.. "High-yield cell-free protein synthesis for site-specific incorporation of unnatural amino acids at two sites" (2012) p. 652 - 656
Available at: http://works.bepress.com/nick_dixon/30/