N-terminally extended proteins expressed in yeast

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/81 (2006.01) C07K 14/62 (2006.01) C12N 15/11 (2006.01) C12N 15/62 (2006.01) C12P 21/06 (2006.01)

Patent

CA 2192943

The present invention relates to polypeptides expressed and processed in yeast, a DNA construct comprising a DNA sequence encoding polypeptide having the following structure signal peptide-leader peptide -X1-X2-X3-X4-X5-X6-X7- heterologous protein, X1 is Lys or Arg; X2 is Lys or Arg, X1 and X2 together defining a yeast processing site; X3 is Glu or Asp; X4 is a sequence of amino acids with the following structure (A-B)n wherein A is Glu or Asp, B is Ala, Val, Leu or Pro, and n is 0 or an integer from 1 to 5, and when n > 2 each A and B is the same or different from the other A(s) and B(s), or X4 is a sequence of amino acids with the following structure (C)m wherein C is Glu or Asp, and m is 0 or an integer from 1 to 5; X5 is a peptide bond or is one or more amino acids which may be the same or different; X6 is a peptide bond or an amino acid residue selected from the group consisting of Pro, Asp, Thr, Ser, Glu, Ala and Gly; and X7 is Lys or Arg.

La présente invention concerne des polypeptides exprimés et transformés par une levure, et des ADN de recombinaison comprenant une séquence d'ADN codant pour des polypeptides ayant la structure suivante: peptide signal - peptide leader -X¿1?-X¿2?-X¿3?-X¿4?-X¿5?-X¿6?-X¿7?- protéine hétérologue. Dans cette formule X¿1? est Lys ou Arg; X¿2? est Lys ou Arg, X¿1? et X¿2? constituant ensemble un site de transformation par la levure; X¿3? est Glu ou Asp; X¿4? est une séquence d'acides aminés ayant la structure (A-B)n, où A est Glu ou Asp, B est Ala, Val, Leu ou Pro et n est égal à 0 ou à un nombre entier entre 1 et 5 et lorsque n > 2, A et B sont identiques ou différents des autres A(s) et B(s); X¿4? est une séquence d'acides aminés ayant la structure (C)m, où C est Glu ou Asp et m est égal à 0 ou à un nombre entier entre 1 et 5; X¿5? est une liaison peptidique ou un ou plusieurs acides aminés qui peuvent être identiques ou différents; X¿6? est une liaison peptidique ou un acide aminé choisi dans le groupe constitué par Pro, Asp, Thr, Ser, Glu, Ala, et Gly; et X¿7? est Lys ou Arg.

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