Expression of polypeptides in yeast

  • US 5,618,676 A
  • Filed: 06/07/1995
  • Issued: 04/08/1997
  • Est. Priority Date: 02/25/1981
  • Status: Expired due to Term
First Claim
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1. A process of forming a transformant of a given yeast strain, which transformant is capable of expressing a biocompetent polypeptide ordinarily exogenous to yeast and not required for growth of said strain, which comprises:

  • (a) providing a DNA transfer vector having bacterial and yeast origins of replication and genes for phenotypic selection of both bacterial and yeast moieties transformed with said genes;

    (b) providing a DNA fragment comprising a structural gene encoding said biocompetent polypeptide;

    (c) providing a DNA fragment comprising a yeast promoter genetically distinct from said structural gene which yeast promoter is from within the about 1500 bp DNA sequence 5'"'"' flanking the start codon of a yeast structural gene;

    (d) inserting the fragments of steps (b) and (c) into said transfer vector together with appropriately positioned translation start and stop signals for said structural gene for said biocompetent polypeptide to form an expression vector in which said structural gene for said biocompetent polypeptide is under the control of said promoter, while maintaining said origins of replications and genes for phenotypic selection, wherein the promoter is resected free of the yeast structural gene and the gene encoding the polypeptide is located at the endpoint of the resection; and

    (e) transforming said strain with the resulting expression vector, wherein said yeast strain is Saccharomyces cerevisiae.

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