Host cells expressing recombinant human erythropoietin

Details Host cells expressing recombinant human erythropoietin Host cells expressing recombinant human erythropoietin

C - Chemistry, Metallurgy

12

P

C12P 21/02 (2006.01) C07K 14/505 (2006.01) C12N 5/10 (2006.01) C12N 15/18 (2006.01) C12N 15/85 (2006.01)

Patent

CA 2350335

The gene coding for human erythropoietin (EPO) was obtained from human genomic DNA. The gene used does not include sequences from regions at i 5' of the first translated ATG and ii 3' of the stop codon of the EPO gene. The gene was cloned into an expression plasmid for eukaryotic cells that have as sole expression control elements the early promoter of the SV40 virus and its polyadenylation signal. Recombinant cells resulting from transfection with genetic constructs used provide an unexpectedly high level of protein expression of 50 mg of recombinant EPO per liter of culture medium per day.

Le gène codant pour l'érythropoïétine humaine (EPO) a été obtenu à partir d'ADN génomique humain. Le gène utilisé ne contient pas les séquences des régions i) 5' du premier ATG transcrit et ii) 3' du codon d'arrêt du gène EPO. Le gène a été cloné en plasmide d'expression pour cellules eucaryotes dont les seuls éléments de régulation d'expression sont le promoteur précoce du virus SV 40 et son signal de polyadénylation. Les cellules recombinantes résultant de la transfection avec les constructions génétiques utilisées offrent un niveau exceptionnellement élevé d'expression de protéine de 50 mg d'EPO recombinante par litre de milieu de culture et par jour.

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