CA2498636A1 - Use of saccharomyces cerevisiae erg4 mutants for the expression of glucose transporters from mammals - Google Patents
Use of saccharomyces cerevisiae erg4 mutants for the expression of glucose transporters from mammals Download PDFInfo
- Publication number
- CA2498636A1 CA2498636A1 CA002498636A CA2498636A CA2498636A1 CA 2498636 A1 CA2498636 A1 CA 2498636A1 CA 002498636 A CA002498636 A CA 002498636A CA 2498636 A CA2498636 A CA 2498636A CA 2498636 A1 CA2498636 A1 CA 2498636A1
- Authority
- CA
- Canada
- Prior art keywords
- yeast cell
- protein
- yeast
- polynucleotide
- compound
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/575—Hormones
- C07K14/62—Insulins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/18—Baker's yeast; Brewer's yeast
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Diabetes (AREA)
- Biotechnology (AREA)
- Mycology (AREA)
- Biochemistry (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- General Engineering & Computer Science (AREA)
- Endocrinology (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Emergency Medicine (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Obesity (AREA)
- Gastroenterology & Hepatology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Botany (AREA)
- Toxicology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Hematology (AREA)
- General Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to yeast strains in which a human GLUT4 transporter or a human GLUT1 transporter can be functionally expressed and in particular GLUT4 transport proteins which can be particularly easily functionally expressed in yeast strains.
Claims (36)
1. A purified and isolated polynucleotide, which comprises a DNA sequence coding for a protein GLUT4V85M.
2. The polynucleotide as claimed in claim 1, which comprises a sequence from any of the following groups:
a) a nucleotide sequence according to Seq ID No. 1 b) a nucleotide sequence which hybridizes to a sequence of Seq ID No. 1 under stringent conditions and which codes for a protein GLUT4V85M.
a) a nucleotide sequence according to Seq ID No. 1 b) a nucleotide sequence which hybridizes to a sequence of Seq ID No. 1 under stringent conditions and which codes for a protein GLUT4V85M.
3. The polynucleotide as claimed in claim 1 or 2, wherein the protein has an amino acid sequence according to Seq ID No. 2.
4. The polynucleotide as claimed in claims 1 to 3, in which the coding region for the protein GLUT4V85M is operationally linked to a promotor.
5. The polynucleotide as claimed in claim 1 to 4, which can be replicated in a yeast cell.
6. The polynucleotide as claimed in claim 5, which can be used to express a protein in a yeast cell.
7. A yeast cell from Saccharomyces cerevisiae, wherein all glucose transporters are no longer functional and which contains no functional Erg4 protein.
8. A yeast cell from Saccharomyces cerevisiae, wherein all glucose transporters are no longer functional and which contains no functional Fgy1 protein and no functional Erg4 protein.
9. The yeast cell as claimed in claim 7 or 8, wherein the ERG4 gene is completely or partially deleted.
10. The yeast cells as claimed in claim 7, as deposited as Saccharomyces cerevisiae DSM 15187.
11. The yeast cells as claimed in claim 8 or 9, as deposited as Saccharomyces cerevisiae DSM 15184.
12. The use of a yeast cell as claimed in claims 15 to 18 for expressing a mammalian GLUT1 protein or GLUT4 protein.
13. The use as claimed in claim 12, for expressing a human GLUT4 protein or a human GLUT1 protein.
14. The yeast cell as claimed in claim 7, comprising a polynucleotide as claimed in claims 1 to 6.
15. The yeast cell as claimed in claim 14, comprising a protein GLUT4V85M.
16. The yeast cell as claimed in claim 14 and/or 15, as deposited as Saccharomyces cerevisiae DSM 15185.
17. A process of preparing a yeast cell as claimed in claims 14 to 16, which comprises the steps:
a) providing a yeast cell as claimed in claim 7, b) providing a polynucleotide as claimed in claim 5 or 6, c) transforming the yeast cell as claimed in a) with the polynucleotide as claimed in b), d) selecting a transformed yeast cell, e) where appropriate, expressing a protein GLUT4V85M.
a) providing a yeast cell as claimed in claim 7, b) providing a polynucleotide as claimed in claim 5 or 6, c) transforming the yeast cell as claimed in a) with the polynucleotide as claimed in b), d) selecting a transformed yeast cell, e) where appropriate, expressing a protein GLUT4V85M.
18. The yeast cell as claimed in claim 8 or 9, comprising a polynucleotide as claimed in claims 1 to 6.
19. The yeast cells claimed in claim 18, comprising a protein GLUT4V85M.
20. The yeast cell as claimed in claim 18 and/or 19, deposited as Saccharomyces cerevisiae DSM 15186.
21. A process of preparing a yeast cell as claimed in claims 18 to 20, which comprises the steps:
a) providing a yeast cell as claimed in claim 8 or 9, b) providing a polynucleotide as claimed in claim 5 or 6, c) transforming the yeast cell as claimed in a) with the polynucleotide as claimed in b), d) selecting a transformed yeast cell, e) where appropriate, expressing a protein GLUT4V85M.
a) providing a yeast cell as claimed in claim 8 or 9, b) providing a polynucleotide as claimed in claim 5 or 6, c) transforming the yeast cell as claimed in a) with the polynucleotide as claimed in b), d) selecting a transformed yeast cell, e) where appropriate, expressing a protein GLUT4V85M.
22. A yeast cell whose glucose transporters in their entirety are no longer functional, comprising a polynucleotide as claimed in claims 1 to 6.
23. The yeast cell as claimed in claim 22, comprising a protein GLUT4V85M.
24. The yeast cell as claimed in claim(s) 22 and/or 23, deposited as Saccharomyces cerevisiae DSM 15188.
25. A process of preparing a yeast cell as claimed in claims 22 to 24, which comprises the steps:
a) producing a yeast cell whose glucose transporters in their entirety are no longer functional, b) providing a polynucleotide as claimed in claim 5 or 6 c) transforming the yeast cell as claimed in a) with the polynucleotide as claimed in b) d) selecting a transformed yeast cell, e) where appropriate, expressing a protein GLUT4V84M.
a) producing a yeast cell whose glucose transporters in their entirety are no longer functional, b) providing a polynucleotide as claimed in claim 5 or 6 c) transforming the yeast cell as claimed in a) with the polynucleotide as claimed in b) d) selecting a transformed yeast cell, e) where appropriate, expressing a protein GLUT4V84M.
26. A protein having the functional activity of a glucose transporter, which is encoded by a polynucleotide sequence as claimed in any of claims 1 to 3.
27. The protein as claimed in claim 13, comprising an amino acid sequence according to Seq. ID No. 2.
28. A method for identifying a compound which stimulates the activity of a protein, which comprises the steps:
a) providing a yeast cell as claimed in one or more of claims 14 to 17, b) providing a chemical compound, c) contacting the yeast of a) with the chemical compound of b), d) determining glucose uptake by the yeast of c), e) relating the detected value of the glucose uptake of d) to the detected value of glucose uptake in a yeast cell as claimed in a) which is not contacted with a chemical compound as claimed in b), with a compound which causes an increase in the amount of glucose taken up in the yeast as claimed in d) stimulating the activity of said GLUT4 protein.
a) providing a yeast cell as claimed in one or more of claims 14 to 17, b) providing a chemical compound, c) contacting the yeast of a) with the chemical compound of b), d) determining glucose uptake by the yeast of c), e) relating the detected value of the glucose uptake of d) to the detected value of glucose uptake in a yeast cell as claimed in a) which is not contacted with a chemical compound as claimed in b), with a compound which causes an increase in the amount of glucose taken up in the yeast as claimed in d) stimulating the activity of said GLUT4 protein.
29. A pharmaceutical comprising a compound, which is identified by means of a method as claimed in claim 28, and additives and excipients for formulating a pharmaceutical.
30. The use of a compound which has been identified by means of a method as claimed in claim 28 for preparing a pharmaceutical for the treatment of type I
and/or II diabetes.
and/or II diabetes.
31. A method for identifying a compound inhibiting the corresponding protein of the Fgy1 gene, which comprises the steps:
a) providing a yeast cell as claimed in one or more of claims 7 to 10 which contains a GLUT 4 protein, b) providing a chemical compound c) contacting the yeast of a) with the chemical compound of b), d) determining glucose uptake by the yeast of c), e) relating the detected value of the glucose uptake of d) to the detected value of glucose uptake in a yeast cell as claimed in a) which is not contacted with a chemical compound as claimed in b), with a compound which causes an increase in the amount of glucose taken up in the yeast as claimed in d) stimulating the activity of a protein Fgy1.
a) providing a yeast cell as claimed in one or more of claims 7 to 10 which contains a GLUT 4 protein, b) providing a chemical compound c) contacting the yeast of a) with the chemical compound of b), d) determining glucose uptake by the yeast of c), e) relating the detected value of the glucose uptake of d) to the detected value of glucose uptake in a yeast cell as claimed in a) which is not contacted with a chemical compound as claimed in b), with a compound which causes an increase in the amount of glucose taken up in the yeast as claimed in d) stimulating the activity of a protein Fgy1.
32. A pharmaceutical comprising a compound which has been identified by means of a method as claimed in claim 31, and additives and excipients for formulating a pharmaceutical.
33. The use of a compound which has been identified by means of a method as claimed in claim 31 for preparing a pharmaceutical for the treatment of diabetes.
34. A method for identifying a compound which inhibits the protein encoded by the ERG4 gene, which method comprises the steps:
a) providing a yeast cell as claimed in one or more of claims 22 to 25, b) providing a chemical compound c) contacting the yeast of a) with the chemical compound of b), d) determining glucose uptake by the yeast of c), e) relating the detected value of the glucose uptake of d) to the detected value of glucose uptake in a yeast cell as claimed in a) which is not contacted with a chemical compound as claimed in b), with a compound which causes an increase in the amount of glucose taken up in the yeast as claimed in d) inhibiting the activity of a protein Erg4.
a) providing a yeast cell as claimed in one or more of claims 22 to 25, b) providing a chemical compound c) contacting the yeast of a) with the chemical compound of b), d) determining glucose uptake by the yeast of c), e) relating the detected value of the glucose uptake of d) to the detected value of glucose uptake in a yeast cell as claimed in a) which is not contacted with a chemical compound as claimed in b), with a compound which causes an increase in the amount of glucose taken up in the yeast as claimed in d) inhibiting the activity of a protein Erg4.
35. A pharmaceutical comprising a compound which has been identified by means of a method as claimed in claim 34, and additives and excipients for formulating a pharmaceutical.
36. The use of a compound which has been identified by means of a method as claimed in claim 34 for preparing a pharmaceutical for the treatment of diabetes.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE10242763A DE10242763A1 (en) | 2002-09-14 | 2002-09-14 | New polynucleotide encoding mutant human glucose transporter, useful for identifying antidiabetic agents that can be used to treat diabetes types I or II |
| DE10242763.1 | 2002-09-14 | ||
| PCT/EP2003/009812 WO2004026907A2 (en) | 2002-09-14 | 2003-09-04 | Use of saccharomyces cerevisiae erg4 mutants for the expression of glucose transporters from mammals |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2498636A1 true CA2498636A1 (en) | 2004-04-01 |
| CA2498636C CA2498636C (en) | 2012-04-17 |
Family
ID=31724774
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2498636A Expired - Fee Related CA2498636C (en) | 2002-09-14 | 2003-09-04 | Use of saccharomyces cerevisiae erg4 mutants for the expression of glucose transporters from mammals |
Country Status (15)
| Country | Link |
|---|---|
| EP (1) | EP1539958A2 (en) |
| JP (1) | JP4520854B2 (en) |
| KR (1) | KR101233998B1 (en) |
| CN (1) | CN1694960B (en) |
| AU (1) | AU2003264257B2 (en) |
| BR (1) | BR0314115A (en) |
| CA (1) | CA2498636C (en) |
| DE (1) | DE10242763A1 (en) |
| HK (1) | HK1084401A1 (en) |
| IL (2) | IL167321A (en) |
| MX (1) | MXPA05002816A (en) |
| NO (1) | NO20051795L (en) |
| RU (1) | RU2345136C2 (en) |
| WO (1) | WO2004026907A2 (en) |
| ZA (1) | ZA200501871B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102899328A (en) * | 2012-09-28 | 2013-01-30 | 南京农业大学 | Goat glucose transporter 4 gene as well as recombinant expression vector and application of gene |
| CN110408616B (en) * | 2019-07-09 | 2021-06-15 | 中南民族大学 | GLUT4 gene knockout sgRNA and A549 cell lines and construction method thereof |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5942398A (en) * | 1998-02-26 | 1999-08-24 | Millennium Pharmaceuticals, Inc. | Nucleic acid molecules encoding glutx and uses thereof |
| EP1189943A1 (en) * | 1999-06-09 | 2002-03-27 | Whitehead Institute For Biomedical Research | Method of measuring plasma membrane targeting of glut4 |
| DE10106718A1 (en) * | 2001-02-14 | 2002-09-05 | Aventis Pharma Gmbh | Yeast strain of Saccharomyces cerevisiae with functional expression of a glow transporter |
-
2002
- 2002-09-14 DE DE10242763A patent/DE10242763A1/en not_active Withdrawn
-
2003
- 2003-09-04 CN CN038251485A patent/CN1694960B/en not_active Expired - Fee Related
- 2003-09-04 EP EP03797264A patent/EP1539958A2/en not_active Withdrawn
- 2003-09-04 CA CA2498636A patent/CA2498636C/en not_active Expired - Fee Related
- 2003-09-04 BR BR0314115-2A patent/BR0314115A/en not_active IP Right Cessation
- 2003-09-04 KR KR1020057004292A patent/KR101233998B1/en not_active IP Right Cessation
- 2003-09-04 WO PCT/EP2003/009812 patent/WO2004026907A2/en active Application Filing
- 2003-09-04 AU AU2003264257A patent/AU2003264257B2/en not_active Ceased
- 2003-09-04 JP JP2004536979A patent/JP4520854B2/en not_active Expired - Fee Related
- 2003-09-04 RU RU2005110955/13A patent/RU2345136C2/en not_active IP Right Cessation
- 2003-09-04 MX MXPA05002816A patent/MXPA05002816A/en active IP Right Grant
-
2005
- 2005-03-04 ZA ZA2005/01871A patent/ZA200501871B/en unknown
- 2005-03-08 IL IL167321A patent/IL167321A/en not_active IP Right Cessation
- 2005-04-12 NO NO20051795A patent/NO20051795L/en not_active Application Discontinuation
-
2006
- 2006-04-18 HK HK06104586.6A patent/HK1084401A1/en not_active IP Right Cessation
-
2009
- 2009-03-16 IL IL197621A patent/IL197621A/en not_active IP Right Cessation
Also Published As
| Publication number | Publication date |
|---|---|
| IL197621A (en) | 2014-01-30 |
| KR20050056206A (en) | 2005-06-14 |
| ZA200501871B (en) | 2005-10-26 |
| BR0314115A (en) | 2005-07-12 |
| RU2345136C2 (en) | 2009-01-27 |
| IL197621A0 (en) | 2011-08-01 |
| KR101233998B1 (en) | 2013-02-18 |
| RU2005110955A (en) | 2006-01-20 |
| NO20051795L (en) | 2005-06-08 |
| HK1084401A1 (en) | 2006-07-28 |
| WO2004026907A2 (en) | 2004-04-01 |
| JP2006517088A (en) | 2006-07-20 |
| AU2003264257B2 (en) | 2010-05-20 |
| MXPA05002816A (en) | 2005-05-27 |
| DE10242763A1 (en) | 2004-03-18 |
| CN1694960A (en) | 2005-11-09 |
| JP4520854B2 (en) | 2010-08-11 |
| CA2498636C (en) | 2012-04-17 |
| WO2004026907A3 (en) | 2004-11-11 |
| IL167321A (en) | 2013-11-28 |
| CN1694960B (en) | 2010-05-12 |
| AU2003264257A1 (en) | 2004-04-08 |
| EP1539958A2 (en) | 2005-06-15 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| MKLA | Lapsed |
Effective date: 20150904 |