Expression system

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/81 (2006.01) C07K 14/39 (2006.01) C12P 21/02 (2006.01)

Patent

CA 2684650

The present invention relates to methods for increasing the secretion of a protein of interest (POI) from a eukaryotic cell comprising co-expression of a POI and of at least one protein that enhances protein secretion, said enhancing protein being selected from the group consisting of BMH2, BFR2, C0G6, C0Y1, CUP5, IMH 1, KIN2, SEC31, SSA4 and SSE1. The invention further relates to a yeast promoter sequence, in particular to a promoter sequence of the PET9 gene of P. pastoris, having, under comparable conditions, an increased promoter activity relative to a promoter sequence of the GAP protein. The invention further relates to an expression vector comprising such a promoter sequence and to the use of such an expression vector for expression of a POI in a host cell. The invention further relates to new yeast promoter sequences of genes from P. pastoris, which are useful for expression of a POI in yeast.

La présente invention concerne des procédés d'augmentation de la sécrétion d'une protéine d'intérêt (POI) par une cellule eucaryote, qui comprennent la co-expression d'une POI et d'au moins une protéine qui stimule la sécrétion protéique, ladite protéine stimulante étant choisie dans le groupe constitué par BMH2, BFR2, C0G6, C0Y1, CUP5, IMH 1, KIN2, SEC31, SSA4 et SSE1. L'invention concerne également une séquence promotrice de levure, en particulier une séquence promotrice du gène PET9 de P. pastoris, ayant, dans des conditions comparables, une activité promotrice accrue relativement à une séquence promotrice de la protéine GAP. De plus, l'invention concerne un vecteur d'expression comprenant une séquence promotrice et l'utilisation d'un tel vecteur d'expression pour l'expression d'une POI dans une cellule hôte. L'invention concerne en outre de nouvelles séquences promotrices de levure de gènes de P. pastoris, qui sont utiles pour l'expression d'une POI chez la levure.

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