WO1988010308A1 - Promoteur de levure induisible par pheromones - Google Patents
Promoteur de levure induisible par pheromones Download PDFInfo
- Publication number
- WO1988010308A1 WO1988010308A1 PCT/US1988/002129 US8802129W WO8810308A1 WO 1988010308 A1 WO1988010308 A1 WO 1988010308A1 US 8802129 W US8802129 W US 8802129W WO 8810308 A1 WO8810308 A1 WO 8810308A1
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- Prior art keywords
- dna
- protein
- polypeptide
- yeast
- promoter
- Prior art date
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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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2468—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1) acting on beta-galactose-glycoside bonds, e.g. carrageenases (3.2.1.83; 3.2.1.157); beta-agarase (3.2.1.81)
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/37—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi
- C07K14/39—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi from yeasts
- C07K14/395—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi from yeasts from Saccharomyces
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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
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
- C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
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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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
- C12N9/2431—Beta-fructofuranosidase (3.2.1.26), i.e. invertase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01023—Beta-galactosidase (3.2.1.23), i.e. exo-(1-->4)-beta-D-galactanase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01026—Beta-fructofuranosidase (3.2.1.26), i.e. invertase
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
- C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/61—Fusion polypeptide containing an enzyme fusion for detection (lacZ, luciferase)
Definitions
- the resulting plasmid is introduced into wild-type a_ cells, which are incubated with alpha factor.
- Alpha factor induces a-type cells to respond; induction of the FUSl promoter leads to induction of transcription of the DNA of interest to which it is fused.
- the result is expression of the FUSl-DNA of interest fusion protein (i.e., a fusion protein which is the FUSl-encoded protein and the protein encoded by the DNA of interest) at substantial levels.
- yeast S. cerevisiae can exist in a diploid or a haploid state; in the former, it has 17 pairs of chromosomes and, in the latter, a single set of chromosomes (one copy of each of the 17 chromosomes) . Under favorable conditions, yeast cells exist in the diploid state and multiply rapidly. Multiplication occurs through mitosis, the process by which a diploid cell duplicates its chromosome pairs and divides into a mother cell and a bud, each of which has two copies of every chromosome.
- Conjugation in S. cerevisiae involves the fusion of haploid cells of opposite mating type, followed by the fusion of nuclei to form a diploid.
- the zygote formed by this process buds off diploid cells capable of vegetative growth. Formation of the zygote requires the coordination of two processes—cell fusion and nuclear fusion. Both processes are initiated by mating phero ones: a. cells produce a.-factor, to which alpha cells specifically respond, and alpha cells produce alpha-factor, to which ⁇ cells specifically respond. Duntze, W.D. et al. , European Journal of Biochemistry, 3_5:357-365 (1973); Wilkinson, L.E. and J.R. Pringle, Experimental Cell Research, 8_9:175-187 (1974).
- a recombinant plasmid which can be used in the method of the present invention, the following components are included: a) a fragment of a plasmid, such as pBR322, which includes the plasmid DNA replication origin, which makes it possible to propagate
- the fus2-l mutation was uncovered in a cross between C52a and JY145 (fusl-483) .
- Several tetrads from this cross produced one ascospore which, when mated to ⁇ 453 lawns, displayed a drastic defect as compared to the response with wild type lawns. (fusl alone displays only a partial defect when mated to ⁇ 453).
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Abstract
Un promoteur de levure est induit par une phéromone appropriée et un procédé d'expression d'un gène voulu en quantités considérables consiste à la placer sous le contrôle du promoteur induisible. De l'ADN de codage d'une protéine voulue est fusionné ou relié à un promoteur de levure induisible par une phéromone, tel que les promoteurs FUS1 ou FUS2, et leur fusion est insérée dans un vecteur permettant de produire un nombre élevé de copies; le produit ainsi obtenu est introduit dans des cellules de levure de type sauvage. La stimulation de ces cellules de levure par la phéromone appropriée entraîne l'induction de la transcriptase du promoteur de levure et l'expression de quantités considérables de la protéine voulue.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
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US6607887A | 1987-06-24 | 1987-06-24 | |
US066,078 | 1987-06-24 |
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WO1988010308A1 true WO1988010308A1 (fr) | 1988-12-29 |
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PCT/US1988/002129 WO1988010308A1 (fr) | 1987-06-24 | 1988-06-23 | Promoteur de levure induisible par pheromones |
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Cited By (17)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1998013513A3 (fr) * | 1996-09-24 | 1998-09-03 | Cadus Pharmaceutical Corp | Procedes et compositions pour identifier des modulateurs de recepteur |
EP0915154A1 (fr) * | 1993-03-31 | 1999-05-12 | Cadus Pharmaceuticals, Inc. | Cellules de levure traítées pour produire des substituts de protéines du système de phéromones, et leurs emplois |
WO1999060138A3 (fr) * | 1998-05-16 | 2000-01-20 | Univ Manchester | Expression regulee de pkc et/ou de srb1/psa1 dans la levure |
US6251605B1 (en) | 1998-10-27 | 2001-06-26 | Cadus Pharmaceutical Corporation | Yeast cells having mutations in Cav1 and uses therefor |
US6255059B1 (en) | 1993-03-31 | 2001-07-03 | Cadus Pharmaceutical Corporation | Methods for identifying G protein coupled receptor effectors |
US6355473B1 (en) | 1998-05-06 | 2002-03-12 | Cadus Pharmaceutical Corp. | Yeast cells having mutations in stp22 and uses therefor |
US6504008B1 (en) | 1999-02-01 | 2003-01-07 | Cadus Technologies, Inc. | Cell based signal generation |
US6555325B1 (en) | 1999-06-14 | 2003-04-29 | Cadus Technologies, Inc. | System for detection of a functional interaction between a compound and a cellular signal transduction component |
US6864060B1 (en) | 1993-03-31 | 2005-03-08 | Cadus Technologies, Inc. | Yeast cells expressing modified G proteins and methods of use therefor |
US7081360B2 (en) | 1998-07-28 | 2006-07-25 | Cadus Technologies, Inc. | Expression of G protein-coupled receptors with altered ligand binding and/or coupling properties |
US7105309B2 (en) | 1993-03-31 | 2006-09-12 | Cadus Technologies, Inc. | Yeast cells engineered to produce pheromone system protein surrogates and uses therefor |
US7223533B2 (en) | 1999-09-10 | 2007-05-29 | Cadus Technologies, Inc. | Cell surface proteins and use thereof as indicators of activation of cellular signal transduction pathways |
US7235648B1 (en) | 1993-03-31 | 2007-06-26 | Cadus Technologies, Inc. | Yeast cells engineered to produce pheromone system protein surrogates, and uses therefor |
US7250263B2 (en) | 1993-03-31 | 2007-07-31 | Cadus Technologies, Inc. | Methods and compositions for identifying receptor effectors |
US7273747B2 (en) | 1998-08-27 | 2007-09-25 | Cadus Technologies, Inc. | Cell having amplified signal transduction pathway responses and uses therefor |
US7319009B2 (en) | 1998-10-07 | 2008-01-15 | Cadus Technologies, Inc. | Methods and compositions for identifying receptor effectors |
US7416881B1 (en) | 1993-03-31 | 2008-08-26 | Cadus Technologies, Inc. | Yeast cells engineered to produce pheromone system protein surrogates, and uses therefor |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0123544A2 (fr) * | 1983-04-25 | 1984-10-31 | Genentech, Inc. | Procédé d'expression de protéines hétérologues dans les levures, vecteurs d'expression et micro-organismes du genre levure à cet effet |
EP0183350A2 (fr) * | 1984-10-29 | 1986-06-04 | Immunex Corporation | ADN codant pour le facteur stimulant des colonies, peptide codé par celui-ci, vecteurs et hôtes transformés contenant un tel ADN et leur préparation |
US4615974A (en) * | 1981-08-25 | 1986-10-07 | Celltech Limited | Yeast expression vectors |
-
1988
- 1988-06-23 WO PCT/US1988/002129 patent/WO1988010308A1/fr unknown
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4615974A (en) * | 1981-08-25 | 1986-10-07 | Celltech Limited | Yeast expression vectors |
EP0123544A2 (fr) * | 1983-04-25 | 1984-10-31 | Genentech, Inc. | Procédé d'expression de protéines hétérologues dans les levures, vecteurs d'expression et micro-organismes du genre levure à cet effet |
EP0183350A2 (fr) * | 1984-10-29 | 1986-06-04 | Immunex Corporation | ADN codant pour le facteur stimulant des colonies, peptide codé par celui-ci, vecteurs et hôtes transformés contenant un tel ADN et leur préparation |
Non-Patent Citations (7)
Title |
---|
CHEMICAL ABSTRACTS, Vol. 103, (1985), Abstract No. 208060x, (MIYAJIMA ATUSUSHI); & GENE, 1985, 37(1-3), 155-61 (Eng). * |
CHEMICAL ABSTRACTS, Vol. 104, (1986), Abstract No. 16037u, (SINGH ARJUN et al.); & GENET: NEW FRONT., PROC. INT. CONGR., 15th 1983, (Pub. 1984). 2, 33-9 (Eng). * |
CHEMICAL ABSTRACTS, Vol. 104, (1986), Abstract No. 220092n, (VLASUK GEORGE P.); & J. BIOL. CHEM., 1986, 261(11), 4789-96 (Eng). * |
CHEMICAL ABSTRACTS, Vol. 107, (1987), Abstract No. 128287u, (McCAFFREY); & MOL. CELL. BIOL., 1987, 7(8), 2680-90 (Eng). * |
CHEMICAL ABSTRACTS, Vol. 107, (1987), Abstract No. 212743x, (TRUEHEART); & MOL. CELL. BIOL., 1987, 7(7), 2316-28, (Eng). * |
CHEMICAL ABSTRACTS, Vol. 108, (1988), Abstract No. 217221f, (NONATO, ROBERTO V.); & BIOCHEM. BIOPHYS. RES. COMMUN., 1988, 152(1), 76-82 (Eng). * |
MOLECULAR AND CELLULAR BIOLOGY, Volume 7, No. 2, February 1987, SCOTT W. VAN ARSDELL et al., "The Yeast Repeated Element Sigma Contains a Hormone-Inducible Promoter", pages 749-759. * |
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US7361498B2 (en) | 1993-03-31 | 2008-04-22 | Cadus Technologies, Inc. | Yeast cells expressing modified G proteins and methods of use therefor |
US6255059B1 (en) | 1993-03-31 | 2001-07-03 | Cadus Pharmaceutical Corporation | Methods for identifying G protein coupled receptor effectors |
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US7105309B2 (en) | 1993-03-31 | 2006-09-12 | Cadus Technologies, Inc. | Yeast cells engineered to produce pheromone system protein surrogates and uses therefor |
US6864060B1 (en) | 1993-03-31 | 2005-03-08 | Cadus Technologies, Inc. | Yeast cells expressing modified G proteins and methods of use therefor |
US6159705A (en) * | 1996-09-24 | 2000-12-12 | Cadus Pharmaceutical Corporation | Recombinant yeast cells for identifying receptor effectors |
WO1998013513A3 (fr) * | 1996-09-24 | 1998-09-03 | Cadus Pharmaceutical Corp | Procedes et compositions pour identifier des modulateurs de recepteur |
US6355473B1 (en) | 1998-05-06 | 2002-03-12 | Cadus Pharmaceutical Corp. | Yeast cells having mutations in stp22 and uses therefor |
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GB2353798A (en) * | 1998-05-16 | 2001-03-07 | Univ Manchester | Regulated expression of PKC and/or SRB1/PSA1 in yeast |
WO1999060138A3 (fr) * | 1998-05-16 | 2000-01-20 | Univ Manchester | Expression regulee de pkc et/ou de srb1/psa1 dans la levure |
US7081360B2 (en) | 1998-07-28 | 2006-07-25 | Cadus Technologies, Inc. | Expression of G protein-coupled receptors with altered ligand binding and/or coupling properties |
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US7223533B2 (en) | 1999-09-10 | 2007-05-29 | Cadus Technologies, Inc. | Cell surface proteins and use thereof as indicators of activation of cellular signal transduction pathways |
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