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US4615974A
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Yeast expression vectors
Assignee: Vernalis R&D Ltd
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Filing: 1982-08-17
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Granted: 1986-10-07
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Expires: 2002-08-17
Abstract
Abstract There are described a number of plasmid vectors suitable for the expression of genetic material, at various levels in yeasts. The plasmids each comprise a yeast selective marker, a yeast replication origin and a yeast promoter positioned relative to a unique restriction site in such a way that expression may be obtained of a polypeptide coding sequence inserted at the restriction site. The promoters used are derived from the 5' region of a gene coding for a yeast glycolytic enzyme e.g. phosphoglycerate kinase (PGK), or from the 5' region of the yeast TRP1 gene. In one Example a plasmid contains a promoter derived from both the 3' and 5' regions of the PGK gene. The replication systems used involve the yeast 2 mu replication origin or an autonomous replicating sequence (ARS) stabilized with an ARS stabilizing sequence (ASS). The replication systems allow for a choice of high or low copy number per cell. The promoter sequences allow for a choice of high or low expression level. A kit including vectors having a combination of these alternative features is described. Yeast expression vectors including a gene for coding for human interferon- alpha are described.
Inventors
- 👤 Alan J. Kingsman
- 👤 Susan M. Kingsman
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-04 21:29
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/52; C07K14/555; C07K14/56; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/80; C12N15/81; Y; Y10; Y10S; Y10S930/00; Y10S930/01; Y10S930/14; Y10S930/142
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
89 words
A yeast expression vector having a Saccharomyces cerevisiae promoter positioned relative to a non Saccharomyces cerevisiae polypeptide coding sequence in such a way that expression may be obtained of said polypeptide coding sequence wherein said Saccharomyces cerevisiae promoter consists essentially of at least a functionally active promoter portion of the 5' region of the PGK gene or at least a functionally active promoter portion of the 5' region of the TRP1 gene, said yeast expression vector also including a Saccharomyces cerevisiae selective marker and a Saccharomyces cerevisiae replication origin.
Claim 2
Dependent Claim
89 words
A yeast expression vector having a Saccharomyces cerevisiae promoter positioned relative to a non Saccharomyces cerevisiae polypeptide coding sequence in such a way that expression may be obtained of said polypeptide coding sequence wherein said Saccharomyces cerevisiae promoter consists essentially of at least a functionally active promoter portion of the 5' region of the PGK gene or at least a functionally active promoter portion of the 5' region of the TRP1 gene, said yeast expression vector also including a Saccharomyces cerevisiae selective marker and a Saccharomyces cerevisiae replication origin.
Claim 3
Dependent Claim
57 words
A yeast expression vector according to claim 1 wherein said at least a functionally active portion of the 5' region of the yeast PGK gene is located upstream of the polypeptide coding sequence and said at least a functionally active portion of the 3' region of the PGK gene is located downstream of the polypeptide coding sequence.
Claim 4
Dependent Claim
32 words
A yeast expression vector according to claim 1 expression control of which is exercised by varying the level of a fermentable carbon source in a nutrient medium of a yeast transformed therewith.
Claim 5
Dependent Claim
15 words
A yeast expression vector according to claim 3 wherein the fermentable carbon source is glucose.
Claim 6
Dependent Claim
40 words
A yeast expression vector according to claim 1 wherein the yeast expression vector contains at least a functionally active portion of the yeast 2μ plasmid replication origin and at least a functionally active portion of the LEU2 yeast selective marker.
Claim 7
Dependent Claim
27 words
A yeast expression vector according to claim 1 wherein the yeast expression vector contains at least a functionally active portion of an autonomous replicating sequence stabilising sequence.
Claim 8
Dependent Claim
20 words
A yeast expression vector according to claim 1 wherein the yeast expression vector contains a gene coding for human interferon-α.
Claim 9
Dependent Claim
42 words
A method for the production of a non Saccharomyces cerevisiae polypeptide comprising expressing a coding sequence for the said polypeptide in a Saccharomyces cerevisiae host organism transformed with a yeast expression vector according to claim 1 containing the said polypeptide coding sequence.
Claim 10
Dependent Claim
12 words
Saccharomyces cerevisiae transformed by a yeast expression vector according to claim 1.
Claim 11
Dependent Claim
56 words
A kit of expression vectors comprising four or more different yeast expression vectors wherein each vector has either of the TRP1:ARS1:ASS or LEU2:2μ replication origin selective marker and replication system and a Saccharomyces cerevisiae promoter consisting essentially of at least a functionally active portion of either of the TRP1 or PGK 5' region Saccharomyces cerevisiae promoter.
Claim 12
Dependent Claim
56 words
A kit of expression vectors comprising four or more different yeast expression vectors wherein each vector has either of the TRP1:ARS1:ASS or LEU2:2μ replication origin selective marker and replication system and a Saccharomyces cerevisiae promoter consisting essentially of at least a functionally active portion of either of the TRP1 or PGK 5' region Saccharomyces cerevisiae promoter.
Claim 13
Dependent Claim
102 words
A kit of yeast expression vectors comprising two or more different yeast expression vectors, each yeast expression vector having a Saccharomyces cerevisiae promoter positioned relative to a Saccharomyces cerevisiae polypeptide coding sequence in such a way that expression may be obtained of said polypeptide coding sequence wherein said Saccharomyces cerevisiae promoter consists essentially of at least a functionally active promoter portion of the 5' region of the PGK gene or at least a functionally active promoter portion of the 5' region of the TRP1 gene, each yeast expression vector also including a Saccharomyces cerevisiae selective marker and a Saccharomyces cerevisiae replication origin.
Claim 14
Dependent Claim
102 words
A kit of yeast expression vectors comprising two or more different yeast expression vectors, each yeast expression vector having a Saccharomyces cerevisiae promoter positioned relative to a Saccharomyces cerevisiae polypeptide coding sequence in such a way that expression may be obtained of said polypeptide coding sequence wherein said Saccharomyces cerevisiae promoter consists essentially of at least a functionally active promoter portion of the 5' region of the PGK gene or at least a functionally active promoter portion of the 5' region of the TRP1 gene, each yeast expression vector also including a Saccharomyces cerevisiae selective marker and a Saccharomyces cerevisiae replication origin.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1989-10-06 | 1990-04-07 | 1990-10-07 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1993-10-06 | 1994-04-07 | 1994-10-07 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1997-10-06 | 1998-04-07 | 1998-10-07 | $7700.00 | Window Closed / Lapsed |