A novel method for selective isotope labeling of bacterially expressed proteins

Karen M. Lee, Elliot Androphy, James D. Baleja

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

A novel method for isotope labeling in selected amino acids is presented for use with the T7 RNA polymerase system. The protocol is illustrated with the DNA-binding domain from the E2 protein of bovine papillomavirus, BPV-1. On addition of rifampicin, protein expression occurs exclusively from the gene controlled by the T7 promoter. Since the bacteria are now dedicated to the production of E2 protein, labeling with specific amino acids is efficiently performed. For example, 10 mg/l of 15N-labeled phenylalanine is shown to be sufficient for incorporation of the label, without scrambling, and without the use of an auxotrophic strain.

Original languageEnglish (US)
Pages (from-to)93-96
Number of pages4
JournalJournal of Biomolecular NMR
Volume5
Issue number1
DOIs
StatePublished - Jan 1995
Externally publishedYes

Fingerprint

Isotope Labeling
Isotopes
Labeling
Amino Acids
Rifampin
Phenylalanine
Labels
Bacteria
Proteins
Genes
DNA
Bovine papillomavirus E2 protein
bacteriophage T7 RNA polymerase

Keywords

  • E2
  • Papillomavirus
  • Protein expression
  • Rifampicin
  • T7 RNA polymerase

ASJC Scopus subject areas

  • Spectroscopy
  • Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

A novel method for selective isotope labeling of bacterially expressed proteins. / Lee, Karen M.; Androphy, Elliot; Baleja, James D.

In: Journal of Biomolecular NMR, Vol. 5, No. 1, 01.1995, p. 93-96.

Research output: Contribution to journalArticle

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