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Expanding the genetic code of Saccharomyces cerevisiae with methionine analogues by Birgit Wiltschi; Waltraud Wenger; Sebastian Nehring; Nediljko Budisa is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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## Abstract We replaced the single N‐terminal methionine in heterologously expressed human Cu/Zn superoxide dismutase with the non‐canonical methionine analogues homopropargylglycine and norleucine in the yeast __Saccharomyces cerevisiae__. Our non‐canonical amino acid incorporation protocol involves a two‐step procedure. In the first step, the methionine auxotrophic yeast cells are accumulated in synthetic medium containing methionine while the target protein production is shut off. After a short methionine depletion phase, the cells are transferred to inducing medium that contains the methionine analogue instead of methionine and target protein expression is switched on. The initially low level incorporation of ∼12% could be elevated to 40% by increasing the non‐canonical amino acid concentration in the medium by 10‐fold. With this approach we were able to produce up to 5 mg substituted protein per litre of yeast culture. Copyright © 2008 John Wiley & Sons, Ltd.

Who reads Expanding the genetic code of Saccharomyces cerevisiae with methionine analogues?

Author

Birgit Wiltschi; Waltraud Wenger; Sebastian Nehring; Nediljko Budisa

Publisher

John Wiley and Sons; Wiley (Blackwell Publishing); John Wiley & Sons Inc.; [Chichester]; [New York]: John Wiley & Sons, ©1985-; Wiley; Hindawi Limited (ISSN 0749-503X)

Published

2008

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)