Cost-effective large-scale expression of proteins for NMR studies
| Autor: | Julia Klopp, Daniela Scherer-Becker, Aurélie Winterhalter, Christian Ostermeier, Rémy Gébleux, Alvar D. Gossert |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
010402 general chemistry medicine.disease_cause 01 natural sciences Biochemistry Protein expression 03 medical and health sciences chemistry.chemical_compound Methionine medicine Escherichia coli Nuclear Magnetic Resonance Biomolecular Spectroscopy Gram Shake flask Carbon Isotopes Nitrogen Isotopes Chemistry Scale (chemistry) Reproducibility of Results Deuterium 0104 chemical sciences 030104 developmental biology Auto induction Isotope Labeling Fermentation |
| Zdroj: | Journal of biomolecular NMR. 71(4) |
| ISSN: | 1573-5001 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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