Expression of polypeptides in yeast
AssigneesWashington Research Foundation, Genentech, Inc.
InventorsRonald A. Hitzeman, Franklin E. Hagie IV, Benjamin D. Hall, Gustav Ammerer
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Abstract
DNA expression vectors capable, in a transformant strain of yeast, of expressing a polypeptide under the control of a genetically distinct yeast promoter, processes of forming transformant strains of yeast and transformed yeast strains are disclosed.
Claims38
A process of forming a transformant of a yeast strain capable of expressing a polypeptide ordinarily exogenous to yeast and not required for growth of said strain, comprising transforming said yeast strain with a DNA transfer vector that comprises:
- (a) bacterial and yeast origins of replication and genes for phenotypic selection of both bacterial and yeast moieties transformed with said vector;
- (b) a DNA fragment comprising a structural gene encoding said polypeptide;
- (c) a yeast promoter genetically distinct from said structural gene obtained from the about 1500 bp DNA sequence 5' flanking the start signal of a yeast structural gene;
- and
- (d) translation start and stop signals for said structural gene encoding said polypeptide that are positioned so that said structural gene encoding said polypeptide is under the control of said promoter while maintaining said origins of replications and genes for phenotypic selection.
A DNA expression vector, capable in a transformant strain of yeast of expressing a polypeptide ordinarily exogenous to yeast and not required for growth of said strain, said vector comprising bacterial and yeast origins of replication and a yeast promoter from the about 1500 bp DNA sequence 5' flanking the start signal of a yeast structural gene, said yeast promoter being capable of directing the expression of a gene encoding said polypeptide.
A yeast strain comprising a gene encoding a polypeptide ordinarily exogenous to yeast and not required for growth of said strain under the control of a yeast promoter from the about 1500 bp DNA sequence 5' flanking the start signal of a yeast structural gene.
A yeast strain capable of expressing a polypeptide ordinarily exogenous to yeast and not required for growth of said strain produced by a process comprising transforming said yeast strain with a DNA transfer vector comprising:
- (a) bacterial and yeast origins of replication and genes encoding polypeptides for phenotypic selection of both bacterial and yeast moieties transformed with said vector;
- (b) a DNA fragment comprising a structural gene encoding said polypeptide;
- (c) a DNA fragment comprising a yeast promoter from the about 1500 bp DNA sequence 5' flanking the start signal of a yeast structural gene;
- (d) translation start and stop signals for said structural gene encoding said polypeptide positioned such that said structural gene for said polypeptide is under the control of said yeast promoter, while maintaining said origins of replication and genes encoding polypeptides for phenotypic selection.
Dependencies are inferred from the claim text.