Secretion of hirudin derivatives

C - Chemistry – Metallurgy – 12 – N

Patent

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530/7.2, 195/1.2

C12N 15/15 (2006.01) C07K 14/815 (2006.01) C12N 9/10 (2006.01)

Patent

CA 2038888

A process for obtaining hirudin derivatives from E. coli secretor mutants which entails: (1) construction of a recombinant vector on which there is located the gene coding for a hirudin derivative directly downstream of a DNA section which codes for a bacterial signal peptide; (2) transformation of an E. coli secretor mutant with the recombinant vector constructed in step (1); (3) cultivation of the transformed cells in a medium; and (4) obtaining the hirudin derivative from the medium; and a recombinant vector which contains one or more copies of a gene construct which codes for a protein consisting of a bacterial signal peptide and of a hirudin derivative, and a hirudin derivative with the N-terminal amino-acid sequence A - (SEQ ID NO: 5) in which A represents Ala, Gln, His, Phe, Tyr, Glu, Ser, Asp or Asn.

Un procédé pour obtenir des dérivés d'hirudine à partir des mutants qui secrètent des bactéries E. coli et qui comprend: (1) la construction d'un vecteur recombinant sur lequel se trouve le codage génétique d'un dérivé d'hirudine directement vers l'aval d'une section d'ADN qui produit un code pour un peptide signal bactérien; (2) la transformation d'un mutant qui secrète des bactéries E. coli avec le vecteur recombinant construit à l'étape (1); (3) la culture des cellules transformées dans un milieu; et (4) la cueillette du dérivé d'hirudine à partir du milieu; et un vecteur recombinant qui contient une ou plusieurs copies d'une construction de gènes qui produit un code pour une protéine consistant en un peptide signal bactérien et d'un dérivé d'hirudine, et un dérivé d'hirudine avec une séquence A d'acides aminés N-terminal - (SEQ ID NO: 5) dans laquelle A représente Ala, Gln, His, Phe, Tyr, Glu, Ser, Asp ou Asn.

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