Novo Nordisk A/s

Knud Vad

Jakob Brandt

Thomas Bo Kjeldsen

Svend Havelund

Per Balschmidt

Thomas Boerglum Kjeldsen

Michi Egel-Mitani

Thomas B&oslash;rglum Kjeldsen

Jacob Brandt

Annette Frost Pettersson

The present invention relates to polypeptides expressed and processed in yeast, a DNA construct comprising a DNA sequence encoding such polypeptides, vectors carrying such DNA fragments and yeast cells transformed with the vectors, as well as a process of producing heterologous proteins in yeast.

N-terminally extended proteins expressed in yeast

The present invention relates to a method for expressing heterologous proteins or polypeptides in yeast by culturing a transformed yeast strain which does not contain a functional antibiotic resistance marker gene.

Method of making proteins in transformed yeast cells

The invention describes a process for making desired polypeptides in yeast. The desired products are expressed as leader bound polypeptides connected by means of a monobasic processing site. The desired polypeptide is cleaved from the leader either in vivo or in vitro by enzymatic cleavage.

Process for making desired polypeptides in yeast

The present invention relates to a novel method for the production of short chain polypeptides, including polypeptides having up to 3 disulfide bonds and/or structures rich in basic amino acid residues, and open structured short chain polypeptides, e.g. glucagon, glucagon like peptides and their functional analogues, in genetically modified yeast cells, said genetically modified yeast cells, and a method for the preparation of said yeast cells.

Method for the production of polypeptides

The present invention relates to a DNA expression cassette comprising the following sequence:

Synthetic leader peptide sequences

The present invention relates to a DNA expression cassette comprising the following sequence: 5'-P-SP-LS-PS-*gene*-(T).sub.i -3' wherein P is a promoter sequence, SP is a DNA sequence encoding a signal peptide, LS is a DNA sequence encoding a leader peptide of formula I: GinProIle(Asp/Glu)(Asp/Glu)X.sup.1 (Glu/Asp)X.sup.2 AsnZ(Thr/Ser)X.sup.3 (SEQ ID NO: 77) (I) wherein X.sup.1 is a peptide bond or a codable amino acid; X.sup.2 is a peptide bond, a codable amino acid or a sequence of up to 4 codable amino acids which may be the same or different; Z is a codable amino acid except Pro; and X.sup.3 is a sequence of from 4 to 30 codable amino acids which may be the same or different; PS is a DNA sequence encoding a processing site; *gene* is a DNA sequence encoding a polypeptide; T is a terminator sequence; and i is 0 or 1.

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Vanløse, Denmark

Knud Vad