C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 15/70 (2006.01) C07K 14/57 (2006.01) C12P 21/02 (2006.01)
Patent
CA 2197037
The invention relates to a method of manufacturing soluble recombinant proteins from bacteria, in which the plasmid contains the entire gene behind a promoter, this plasmid being expressed and the protein obtained being isolated and purified, in which the cell culture is raised to an OD 600 nm of 0.4 to 0.6; the cell culture is cooled over a period of 15 to 40 minutes to 20 to 26°C; synthesis is induced by addition of 10 to 55 µM of an inductor; and the induction phase is carried out at a temperature of 20 to 26°C over a period of 3.5 to 6.5 hours.
L'invention porte sur un système de production de protéines solubles recombinantes à partir de bactéries, dans lequel le plasmide renferme le gène entier derrière un promoteur, lequel plasmide est exprimé et la protéine obtenue est isolée et purifiée; dans lequel la culture cellulaire est élevée jusqu'à un DE de 600 nm de 0,4 à 0,6; la culture cellulaire est refroidie pendant une période de 15 à 40 minutes entre 20 et 26 oC; la synthèse est induite par l'ajout de 10 à 55 n.M d'un inducteur; et la phase d'induction est effectuée à une température de 20 à 26 oC pendant une période de 3,5 à 6,5 heures.
Otto Bernd
Waschutza Gero
Zakaria Hayssam
Fraunhofer-Gesellschaft Zur Forderung Der Angewandten Forschung
Mcfadden Fincham
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