Genetic tools for selective labeling of proteins with alpha-15N-amino acids.
A collection of genetic tools that can be used to manipulate amino acid metabolism in Escherichia coli is described. The set comprises 21 strains of bacteria, each containing a different genetic defect that is closely linked to a selectable transposon marker. These tools can be used to construct strains of E. coli with ideal genotypes for residue-specific, selective labeling of proteins with nearly any 15N-amino acid. By using strains which have been modified to contain the appropriate genetic lesions to control amino acid biosynthesis, dilution of the isotope by endogenous amino acid biosynthesis and scrambling of the label to other types of residues can be avoided.
Department of Physical Chemistry, Roche Research Center, Hoffmann-La Roche Inc., Nutley, NJ 07110, USA.
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Waugh
D000596:Amino Acids / Q000096:biosynthesis*; D001426:Bacterial Proteins / Q000096:biosynthesis*; D001433:Bacteriolysis; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005819:Genetic Markers; D005838:Genotype; D007553:Isotope Labeling / Q000379:methods*; D009154:Mutation; D009587:Nitrogen Isotopes; D014161:Transduction, Genetic
D000596:Amino Acids; D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D005819:Genetic Markers; D009587:Nitrogen Isotopes