Cost-effective large-scale expression of proteins for NMR studies.
J Biomol NMR
; 71(4): 247-262, 2018 Aug.
Article
en En
| MEDLINE
| ID: mdl-29779067
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-ß-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.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Resonancia Magnética Nuclear Biomolecular
/
Marcaje Isotópico
Tipo de estudio:
Health_economic_evaluation
Idioma:
En
Revista:
J Biomol NMR
Asunto de la revista:
BIOLOGIA MOLECULAR
/
DIAGNOSTICO POR IMAGEM
/
MEDICINA NUCLEAR
Año:
2018
Tipo del documento:
Article
País de afiliación:
Suiza