Journal
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
Volume 132, Issue 42, Pages 14819-14824Publisher
AMER CHEMICAL SOC
DOI: 10.1021/ja104609m
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Funding
- Medical Research Council (MRC) UK
- European Research Council (ERC)
- MRC [MC_U105181009, MC_UP_A024_1008] Funding Source: UKRI
- Medical Research Council [MC_UP_A024_1008, MC_U105181009] Funding Source: researchfish
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We report the discovery of a simple system through which variant pyrrolysyl-tRNA synthetase/tRNA(CUA)(Pyl) pairs created in Escherichia coli can be used to expand the genetic code of Saccharomyces cerevisiae. In the process we have solved the key challenges of producing a functional tRNA(CUA)(Pyl) in yeast and discovered a pyrrolysyl-tRNA synthetase/tRNA(CUA)(Pyl) pair that is orthogonal in yeast. Using our approach we have incorporated an alkyne-containing amino acid for click chemistry, an important post-translationally modified amino acid and one of its analogs, a photocaged amino acid and a photo-cross-linking amino acid into proteins in yeast. Extensions of our approach will allow the growing list of useful amino acids that have been incorporated in E. coli with variant pyrrolysyl-tRNA synthetase/tRNA(CUA)(Pyl) pairs to be site-specifically incorporated into proteins in yeast.
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