Patent US3258509A successfully refreshed from Google Patents.
EP0103409A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Glycolytic promoters for regulated protein expression: protease inhibitor
Assignee: Zymogenetics Inc
•
Filing: 1983-08-12
•
Granted: —
•
Expires: 2003-08-12
Abstract
Abstract Promoters associated with expression of specific enzymes in the glycolytic pathway are used for expression of alien DNA, particularly yeast promoters known to provide high enzyme levels of enzymes in the glycolytic pathway are employed for expressing a mammalian protein, such as alpha-1-antitrypsin. The promoters include promoters involved in expression of pyruvate kinase, triose phosphate isomerase, phosphoglucose isomerase, phosphoglycerate mutase, hexokinase 1, hexokinase 2, glucokinase, phosphofructose kinase, and aldolase, as well as the glycolytic regulation gene. Particularly, the glycolytic regulation gene can be used in conjunction with promoters in the glycolytic pathway for regulated production of desired proteins.
Inventors
- 👤 Glenn Hitoshi Kawasaki
- 👤 Richard Grant Woodbury
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-27 23:47
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/80; C12N15/81; A; A61; A61P; A61P11/00; A61P29/00; C07; C07K; C07K14/00; C07K14/81; C07K14/8107; C07K14/811; C07K14/8121; C07K14/8125; A61K; A61K38/00
Claims Analysis
Total Claims: 38
Claim 1
Independent Claim
67 words
An extrachromosomal element capable of replication in yeast and containing a yeast promoter capable of regulating the transcription of a glycolytic protein, which protein is triose phosphate isomerase, pyruvate kinase, .. phosphoglucose isomerase, phosphoglycerate mutase, hexokinase 1, hexokinase 2, glucokinase, phosphofructose kinase, and aldolase, or glycolytic regulation protein, said yeast promoter being followed downstream by other than a gene expressing the protein normally regulated by such promoter.
Claim 2
Dependent Claim
67 words
An extrachromosomal element capable of replication in yeast and containing a yeast promoter capable of regulating the transcription of a glycolytic protein, which protein is triose phosphate isomerase, pyruvate kinase, .. phosphoglucose isomerase, phosphoglycerate mutase, hexokinase 1, hexokinase 2, glucokinase, phosphofructose kinase, and aldolase, or glycolytic regulation protein, said yeast promoter being followed downstream by other than a gene expressing the protein normally regulated by such promoter.
Claim 3
Dependent Claim
16 words
An extrachromosomal element according to Claim 1, having a marker for selection in a yeast host.
Claim 4
Dependent Claim
25 words
An extrachromosomal element according to Claim 1, having a gene expressing a foreign protein downstream from said promoter and under the regulation of said promoter.
Claim 5
Dependent Claim
25 words
An extrachromosomal element according to Claim 1, wherein said promoter is followed by a gene expressing protease inhibitor having substantially the same structure as alpha-I-antitrypsin.
Claim 6
Dependent Claim
27 words
An extrachromosomal element according to any of Claims 1, 2, 3 or 4, wherein said promoter is the triose phosphate isomerase promoter or the pyruvate kinase promoter.
Claim 7
Dependent Claim
22 words
An extrachromosomal element according to Claim 5, wherein a gene-expressing an enzyme in a metabolic path way of said yeast is present.
Claim 8
Dependent Claim
17 words
A yeast cell containing an extrachromosomal element according to any of Claims 1, 2, 3 or 4.
Claim 9
Dependent Claim
24 words
. A DNA construct comprising a triose phosphate isomerase promoter followed by a gene expressing a foreign protein under the regulation of said promoter.
Claim 10
Dependent Claim
24 words
. A DNA construct comprising a triose phosphate isomerase promoter followed by a gene expressing a foreign protein under the regulation of said promoter.
Claim 11
Dependent Claim
21 words
9 . A DNA construct according to claim 8 , wherein said foreign protein is a protein having protease inhibition activity.
Claim 12
Dependent Claim
24 words
A DNA construct according to Claim 9 , wherein the major sequence of said protein is the same sequence amino acids as human alpha-1-antitrypsin.
Claim 13
Dependent Claim
21 words
A DNA construct comprising a pyruvate kinase promoter followed by a gene expressing a foreign protein under regulation of said protein.
Claim 14
Dependent Claim
21 words
A DNA construct comprising a pyruvate kinase promoter followed by a gene expressing a foreign protein under regulation of said protein.
Claim 15
Independent Claim
43 words
A method for preparing any foreign protein which comprises: introducing into a yeast host an extrachromosomal element according to Claims 1, 2, 3, or 4; and growing said yeast host in an appropriate medium and isolating the protein expressed by said foreign DNA.
Claim 16
Dependent Claim
41 words
A yeast cell containing at least a portion of an extrachromosomal element according to any of Claims 1, 2, 3 or 4 integrated into the genome of said yeast cell, wherein said portion includes at least said promoter and said gene.
Claim 17
Dependent Claim
15 words
A-yeast cell according to Claim 13, wherein said promoter is the triose phosphate isomerase promoter
Claim 18
Dependent Claim
19 words
1 5. A protease inhibitor produced by culturing a yeast cell having an extrachromosomal element according to Claim 4.
Claim 19
Dependent Claim
90 words
16 . A protease inhibitor produced by culturing a yeast cell containing at least a portion of an extrachromosomal element integrated into said yeast cell genome, wherein said portion includes a yeast promoter capable of regulating the transcription of a glycolytic protein, which protein is triose phosphate isomerase, pyruvate kinase, phosphoglucose isomerase, phosphoglycerate mutase, hexokinase 1, hexokinase 2, glucokinase, phosphofructose kinase, and aldolase, or glycolytic regulation protein, and a gene under the transcription regulation of said promoter and expressing a protease inhibitor having substantially the same polypeptide structure as alpha-1-antitrypsin.
Claim 20
Dependent Claim
90 words
16 . A protease inhibitor produced by culturing a yeast cell containing at least a portion of an extrachromosomal element integrated into said yeast cell genome, wherein said portion includes a yeast promoter capable of regulating the transcription of a glycolytic protein, which protein is triose phosphate isomerase, pyruvate kinase, phosphoglucose isomerase, phosphoglycerate mutase, hexokinase 1, hexokinase 2, glucokinase, phosphofructose kinase, and aldolase, or glycolytic regulation protein, and a gene under the transcription regulation of said promoter and expressing a protease inhibitor having substantially the same polypeptide structure as alpha-1-antitrypsin.
Claim 21
Independent Claim
38 words
A method of producing a polypeptide having the protease inhibition activity of mammalian alpha-1-antitrypsin, comprising the step of growing a culture of microorganisms transformed by a DNA transfer vector, said vector comprising a segment coding for mammalian alpha-1-antitrypsin.
Claim 22
Independent Claim
38 words
A method of producing a polypeptide having the protease inhibition activity of mammalian alpha-1-antitrypsin, comprising the step of growing a culture of microorganisms transformed by a DNA transfer vector, said vector comprising a segment coding for mammalian alpha-1-antitrypsin.
Claim 23
Dependent Claim
14 words
18 . A method according to Claim 17 wherein said culture comprises fungal microorganisms.
Claim 24
Dependent Claim
12 words
A method according to Claim 18 wherein said microorganisms comprise yeast cells.
Claim 25
Dependent Claim
18 words
. A method according to Claim 19.. wherein said yeast cells comprise cells of the mutant strain GK-100.
Claim 26
Dependent Claim
24 words
. A method according to Claim 19 wherein said yeast cells contain allelic mutations to GK-100 mutations whereby hyperproduction of said polypeptide is attained.
Claim 27
Dependent Claim
18 words
A method according to Claim 19 wherein said vector comprises 2-micron plasmid DNA, plasmid CATI or plasmid HAT4.
Claim 28
Dependent Claim
21 words
A method according to Claim 17 further comprising the steps of extracting said polypeptide from said culture and purifying said polypeptide.
Claim 29
Dependent Claim
29 words
A method according to Claim 23 further comprising the step of altering a suitable vector to include said segment coding for mammalian alpha-1-antitrypson to form said DNA transfer vector.
Claim 30
Dependent Claim
18 words
A method according to Claim 23 further comprising the step of introducing said DNA transfer vector into yeast.
Claim 31
Independent Claim
21 words
A method according to any of Claims 20 to 30 wherein said polypeptide comprises the amino acid sequence of naturally-occurring mammalian.alpha-1-antitrypsin.
Claim 32
Dependent Claim
16 words
A method according to Claim 31 wherein said polypeptide is the predominant human form of alpha-1-antitrypsin.
Claim 33
Dependent Claim
13 words
A substantially unglycosylated polypeptide characterized by the amino acid sequence of mammalian alpha-1-antitrypsin.
Claim 34
Dependent Claim
13 words
A substantially unglycosylated polypeptide characterized by the amino acid sequence of mammalian alpha-1-antitrypsin.
Claim 35
Dependent Claim
6 words
Substantially pure, substantially unglycosylated mammalian alpha-1-antitrypsin.
Claim 36
Dependent Claim
6 words
Substantially pure, substantially unglycosylated mammalian alpha-1-antitrypsin.
Claim 37
Dependent Claim
21 words
A substantially unglycosylated polypeptide characterized by the amino acid sequence of mammalian alpha-l-antitrypsin for use in the treatment of diseased states.
Claim 38
Dependent Claim
21 words
A substantially unglycosylated polypeptide characterized by the amino acid sequence of mammalian alpha-l-antitrypsin for use in the treatment of diseased states.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
Maintenance schedule is calculated for granted US utility patents.