Cost-effective large-scale expression of proteins for NMR studies

Journal of Biomolecular NMR
Julia KloppAlvar D Gossert

Abstract

We present protocols for high-level expression of isotope-labelled proteins in E. coli in cost-effective ways. This includes production of large amounts of unlabeled proteins and 13C-methyl methionine labeling in rich media, where yields of up to a gram of soluble protein per liter of culture are reached. Procedures for uniform isotope labeling of 2H, 13C and 15N using auto-induction or isopropyl-β-D-1-thiogalactopyranoside-induction are described, with primary focus on minimal isotope consumption and high reproducibility of protein expression. These protocols are based on high cell-density fermentation, but the key procedures are easily transferred to shake flask cultures.

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Citations

Jul 26, 2018·Journal of Biomolecular NMR·Jerome Boisbouvier, Lewis E Kay
Jan 8, 2019·Journal of Biomolecular NMR·Stephan B AzatianMichael P Latham
Mar 5, 2020·Progress in Nuclear Magnetic Resonance Spectroscopy·Stefan Schütz, Remco Sprangers
Mar 20, 2021·Microbial Biotechnology·István VidaAndrás Perczel

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