15N-labeled P22 c2 repressor for nuclear magnetic resonance studies of protein-DNA interactions.
The salmonella phage P22 c2 repressor was produced with 90% 15N isotope labeling of all leucines, using the expression system E. coli W3110 lac IQ¿P 125. The N-terminal DNA-binding domain 1-76 was obtained by chymotrypsin cleavage. Its characterization by biochemical techniques, mass spectrometry, and one- and two-dimensional nuclear magnetic resonance (NMR) showed that highly residue-selective isotope labeling was achieved with the minimal growth medium used. The ability to obtain such isotope labeling opens new avenues for NMR studies of protein-DNA interactions in the P22 operator system.
D004926:Escherichia coli / Q000235:genetics*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D009587:Nitrogen Isotopes; D010957:Plasmids; D011485:Protein Binding; D012097:Repressor Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D012483:Salmonella Phages / Q000235:genetics*; D014157:Transcription Factors / Q000378:metabolism*
D009587:Nitrogen Isotopes; D012097:Repressor Proteins; D014157:Transcription Factors