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

Klopp, Julia and Winterhalter, Aurelie and Gebleux, Remy and Scherer-Becker, Daniela and Ostermeier, Christian and Gossert, Alvar (2018) Cost-effective large-scale expression of proteins for NMR studies. Journal of Biomolecular NMR. pp. 1-16. ISSN 15735001

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.

Item Type: Article
Keywords: Auto-induction Cost-effective High cell-density fermentation Isotope labeling Methionine labelling
Date Deposited: 02 Aug 2018 00:45
Last Modified: 02 Aug 2018 00:45
URI: https://oak.novartis.com/id/eprint/35024

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