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Biotechniques. 2010 Sep;49(3):659-61.

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

Neerathilingam M, Markley JL.

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Biochemistry Department, University of Wisconsin-Madison, Madison, WI 53706, USA.

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.

PMID:: 20854268; [PubMed - indexed for MEDLINE]
PMCID:: PMC3023234

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