Protein glycosylation modification in pichia pastoris

Details Protein glycosylation modification in pichia pastoris Protein glycosylation modification in pichia pastoris

C - Chemistry, Metallurgy

12

N

C12N 9/24 (2006.01) C12N 9/10 (2006.01) C12N 9/12 (2006.01) C12N 15/81 (2006.01) C12P 21/00 (2006.01)

Patent

CA 2490268

The present invention provides genetically engineered strains of methylotrophic yeast including Pichia and especially Pichia pastoris capable of producing proteins with reduced or modified glycosylation. Methods of producing glycoproteins with reduced and/or modified glycosylation using such genetically engineered strains of Pichia are also provided. Vectors, which comprise coding sequences for .alpha.-1,2-mannosidase I, glucosidase II, GlcNAc-tranferase I and mannosidase II or comprising OCH1 disrupting sequence, for transforming methylotrophic yeasts are contemplated by the present invention. Kit for providing the contemplated vectors are also included in this invention.

L'invention concerne des souches de levure méthylotrophique mises au point génétiquement, incluant <i> Pichia</i>, et notamment <i>Pichia pastoris</i>, qui sont capables de produire des protéines à glycosylation réduite ou modifiée. Des procédés de production de glycoprotéines à glycosylation réduite et/ou modifiée utilisant des souches de <i>Pichia</i> mises au point génétiquement sont également décrits. L'invention concerne aussi des vecteurs comprenant des séquences qui codent pour la .alpha.-1,2-mannosidase I, la glucosidase II, la GlcNAc-transférase I et la mannosidase II, ou comportant une séquence disruptive OCH1, qui servent à transformer des levures méthylotrophiques. Une trousse pour produire lesdits vecteurs est également comprise dans l'invention.

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