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Auto-induction medium containing glyphosate for high-level incorporation of unusual aromatic amino acids into proteins.

ABSTRACT: We describe the use of an auto-induction medium containing N-(phosphono-methyl)glycine (glyphosate) as a means for high-level introduction of nonstandard aromatic amino acids into a protein. We illustrate this approach by preparing maltose binding protein (MBP) wherein all eight tryptophan residues have been replaced with 6-fluorotryptophan at an incorporation level of 99.3%. Such a high level of incorporation is important for spectroscopic investigations, in particular 19F NMR, because each species' differing amino acid sequence potentially yields a different peak pattern that complicates spectral analysis.

SUBMITTER: Neerathilingam M

PROVIDER: S-EPMC3023234 | biostudies-literature | 2010 Sep

REPOSITORIES: biostudies-literature

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Auto-induction medium containing glyphosate for high-level incorporation of unusual aromatic amino acids into proteins.

Neerathilingam Muniasamy M Markley John L JL

BioTechniques 20100901 3

We describe the use of an auto-induction medium containing N-(phosphono-methyl)glycine (glyphosate) as a means for high-level introduction of nonstandard aromatic amino acids into a protein. We illustrate this approach by preparing maltose binding protein (MBP) wherein all eight tryptophan residues have been replaced with 6-fluorotryptophan at an incorporation level of 99.3%. Such a high level of incorporation is important for spectroscopic investigations, in particular 19F NMR, because each spe ...[more]

PMID: 20854268

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