Auto-induction medium containing glyphosate for high-level incorporation unusual aromatic amino acids into proteins. (September 2010)

• Record Type: Journal Article
• Authors: Neerathilingam, Muniasamy; Markley, John L.
• Is Part Of: Biotechniques. Volume 49:Number 3(2010)
• Page Start: 659
• Page End: 661
• Publication Date: 2010-09
• Subjects: Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/000113491
• Languages: English
• ISSNs: 0736-6205

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 19 F NMR, because each species— differing amino acid sequence potentially yields a different peak pattern that complicates spectral analysis.