Fgf18 production in prokaryotic hosts

C - Chemistry – Metallurgy – 07 – K

Patent

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C07K 14/50 (2006.01)

Patent

CA 2592261

The expression vectors and methods using an E. coli expression system for the large scale production of FGF18 are described. The vectors utilize the FGF18 coding sequence with specific changes in nucleotides in order to optimize codons and mRNA secondary structure for translation in E. coli. Using the expression vectors, the FGF18 gene was produced in E. coli to a level of greater than 1 g/L in fed batch fermentation. Also included are OmpT deficient E. coli strains, as well as OmpT and fhuA negative strains transformed with an FGF18 expression vector.

La présente invention concerne des vecteurs d'expression et des méthodes dans lesquelles on utilise un système d'expression E. coli pour la production de masse de FGF18. Les vecteurs utilisent la séquence de codage FGF18 ainsi que les changements spécifiques de nucléotides afin d'optimiser les codons et la structure secondaire d'ARNm pour la traduction dans E. coli. Au moyen des vecteurs d'expression, le gène FGF18 a été produit dansE. coli à un niveau supérieur à 1 g/L dans une culture en alimentation programmée. Cette invention concerne également des souches E.coli déficientes en OmpT et des souches négatives fhuA transformées avec un vecteur d'expression FGF18.

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