A recombinant expression vector of human parathyroid hormone

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/16 (2006.01) C07H 21/04 (2006.01) C07K 17/00 (2006.01) C08H 1/00 (2006.01) C12N 1/21 (2006.01) C12N 15/09 (2006.01) C12N 15/70 (2006.01) C12P 21/00 (2006.01) C12P 21/02 (2006.01)

Patent

CA 2298106

The present invention relates to a recombinant expression vector which is prepared by inserting a human parathyroid hormone gene containing a urokinase-specific cleavage site into an L-arabinose inducible vector containing a phosphoribulokinase gene fragment of Rhodobacter sphaeroides or its mutated gene as a fusion partner, a recombinant microorganism transformed with the said expression vector, and a process for preparing human parathyroid hormone on a large scale by cultivating the said microorganism in a medium containing L-arabinose. In accordance with the invention, a recombinant human PTH having the same activity of the native human PTH can be prepared in a high yield through the precise control of induction by a manufacturing process which comprises a step of inducing expression of fusion protein in the microorganism transformed with the recombinant expression vector by L-arabinose.

La présente invention porte sur un vecteur d'expression de recombinaison préparé par introduction d'un gène de la parathormone humaine contenant un site de clivage spécifique de l'urokinase dans un vecteur inductible L-arabinose contenant un fragment du gène phosphoribulokinase de <u>Rhodobacter sphaeroïdes</u> ou son gène muté utilisé comme partenaire de fusion. L'invention porte également sur un micro-organisme de recombinaison transformé par le vecteur d'expression et sur un processus de préparation à grande échelle de la parathormone consistant à mettre en culture le micro-organisme dans un milieu contenant L-arabinose. Selon l'invention, une parathormone humaine de recombinaison ayant la même activité que la parathormone humaine native peut être préparée à des rendements élevés par un contrôle précis d'induction selon un processus de fabrication qui consiste à induire l'expression de la protéine de fusion dans le micro-organisme transformé par le vecteur d'expression de recombinaison par L-arabinose.

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