Recommendations for the Selective Labeling of [15N]-Labeled Amino Acids without Using Auxotrophic Strains

Abstract

The strategy to incorporate [15N]-labeled amino acids were discussed. Instead of using specific auxotrophic strains for selective labeling, the prototrophic strain, BL21(DE3), was used with a plasmid, pLysS, and found to be very effective for several amino acids including alanine, lysine, leucine, and threonine. Isoleucine, valine, glutamine, and tyrosine were also found to be effective despite some diffusion into other amino acids. Interesting result was obtained when [15N]-labeled glycine was tried: only glycines were labeled when amino acid mixture was added in the growth medium, and serines were co-labeled when amino acids were omitted. These results can be used as a guideline when selective labeling strategy is considered, and when the resulting data are interpreted.