Method for the recombinant expression of an n-terminal...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/12 (2006.01) C12N 15/70 (2006.01)

Patent

CA 2558742

A method for the production of a-chain of hepatocyte growth factor or an N- terminal fragment thereof (NK polypeptide) by expression of a nucleic acid encoding said NK polypeptide in a microbial host cell, isolating of inclusion bodies containing said NK polypeptide in denatured form, solubilization of the inclusion bodies and naturation of the denatured NK polypeptide, which is characterized in that in said nucleic acid at least one of the codons of amino acids selected from the group consisting of codons at positions 33, 35 and 36 is CGT, results in an improved expression yield.

L'invention porte sur une méthode d'obtention de la chaîne .alpha. du facteur de croissance de l'hépatocyte ou d'un de ses fragments N-terminaux (polypeptide NK) comportant les étapes suivantes: expression d'un acide nucléique codant pour ledit polypeptide NK dans une cellule hôte microbienne; isolement des corps d'inclusion contenant ledit polypeptide NK sous forme dénaturée; solubilisation des corps d'inclusion; et naturation du polypeptide NK dénaturé. Ladite méthode est caractérisée en ce que dans ledit acide nucléique l'un au moins des codons des acides aminés sélectionnés parmi les codons se trouvant aux positions 33, 35 et 36 est un codon CGT, d'où un rendement d'expression amélioré.

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