CU22722A1 - TRANSFORMATION SYSTEM FOR THE EXPRESSION OF HETEROLOGICAL GENES IN CANDIDA UTILIS YEAST - Google Patents
TRANSFORMATION SYSTEM FOR THE EXPRESSION OF HETEROLOGICAL GENES IN CANDIDA UTILIS YEASTInfo
- Publication number
- CU22722A1 CU22722A1 CU1996082A CU1996082A CU22722A1 CU 22722 A1 CU22722 A1 CU 22722A1 CU 1996082 A CU1996082 A CU 1996082A CU 1996082 A CU1996082 A CU 1996082A CU 22722 A1 CU22722 A1 CU 22722A1
- Authority
- CU
- Cuba
- Prior art keywords
- yeast
- candida utilis
- expression
- host
- genes
- Prior art date
Links
- 240000004808 Saccharomyces cerevisiae Species 0.000 title abstract 6
- 241000235646 Cyberlindnera jadinii Species 0.000 title abstract 5
- 108090000623 proteins and genes Proteins 0.000 title abstract 5
- 230000009466 transformation Effects 0.000 title abstract 4
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 abstract 3
- 238000000034 method Methods 0.000 abstract 3
- 102000004169 proteins and genes Human genes 0.000 abstract 3
- 108020004511 Recombinant DNA Proteins 0.000 abstract 2
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 abstract 2
- 230000006696 biosynthetic metabolic pathway Effects 0.000 abstract 2
- 239000013612 plasmid Substances 0.000 abstract 2
- 101150110971 CIN7 gene Proteins 0.000 abstract 1
- 241000222120 Candida <Saccharomycetales> Species 0.000 abstract 1
- 108020004414 DNA Proteins 0.000 abstract 1
- 102000004190 Enzymes Human genes 0.000 abstract 1
- 108090000790 Enzymes Proteins 0.000 abstract 1
- XYZZKVRWGOWVGO-UHFFFAOYSA-N Glycerol-phosphate Chemical compound OP(O)(O)=O.OCC(O)CO XYZZKVRWGOWVGO-UHFFFAOYSA-N 0.000 abstract 1
- 101150009006 HIS3 gene Proteins 0.000 abstract 1
- 101100246753 Halobacterium salinarum (strain ATCC 700922 / JCM 11081 / NRC-1) pyrF gene Proteins 0.000 abstract 1
- 102000004867 Hydro-Lyases Human genes 0.000 abstract 1
- 108090001042 Hydro-Lyases Proteins 0.000 abstract 1
- 101150110298 INV1 gene Proteins 0.000 abstract 1
- 108091028043 Nucleic acid sequence Proteins 0.000 abstract 1
- 102100037214 Orotidine 5'-phosphate decarboxylase Human genes 0.000 abstract 1
- 108010055012 Orotidine-5'-phosphate decarboxylase Proteins 0.000 abstract 1
- 101100394989 Rhodopseudomonas palustris (strain ATCC BAA-98 / CGA009) hisI gene Proteins 0.000 abstract 1
- 101150050575 URA3 gene Proteins 0.000 abstract 1
- 101100397044 Xenopus laevis invs-a gene Proteins 0.000 abstract 1
- 230000000295 complement effect Effects 0.000 abstract 1
- 230000002950 deficient Effects 0.000 abstract 1
- 239000012634 fragment Substances 0.000 abstract 1
- 230000002068 genetic effect Effects 0.000 abstract 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 abstract 1
- 230000006801 homologous recombination Effects 0.000 abstract 1
- 238000002744 homologous recombination Methods 0.000 abstract 1
- 238000012423 maintenance Methods 0.000 abstract 1
- 238000002703 mutagenesis Methods 0.000 abstract 1
- 231100000350 mutagenesis Toxicity 0.000 abstract 1
- 230000035772 mutation Effects 0.000 abstract 1
- 230000028327 secretion Effects 0.000 abstract 1
- 229940035893 uracil Drugs 0.000 abstract 1
Classifications
-
- 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
-
- 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
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Mycology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- Medicinal Chemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
La presente invención está relacionada con el campo de la biotecnología y la tecnología del ADN recombinante y en particular con un método para la transformación genética reproducible de la levadura Candida utilis, los ADN recombinantes empleados para ello y las cepas de levadura obtenidas. Esta invención también está relacionada con nuevas secuencias de ADN que pudieran ser utilizadas como promotores y terminadores para la expresión de proteínas en esta levadura. Su objetivo técnico es el desarrollo de un sistema hospedero - vector que permita la expresión y la secreción de proteínas de interés en esta levadura. La presente invención proporciona un método para la transformación estable y el mantenimiento del ADN en la cepa hospedera, donde éste es integrado en el genoma por recombinación homóloga. La presente invención proporciona un método para la expresión de genes heterólogos el cual usa como hospedero nuevos mutantes auxotróficos en la levadura Cándida utilis NRRL Y1084 los cuales son defectivos en la enzima orotidina - 5´fosfato descarboxilasa de la vía biosintética del uracilo y una enzima imidazol glicerol fosfato deshidratasa de la vía biosintética de la histidina, respectivamente y fueron obtenidos por mutagénesis de una cepa del género Cándida. Estos mutantes presentan una alta estabilidad y son capaces de ser transformados con alta eficiencia por un vector integrativo del tipo usado en esta invención. Los plásmidos obtenidos a partir de la genoteca fueron utilizados para complementar mutaciones en Cándida utilis. Estos plásmidos contienen el fragmento asociado a los genes URA3, HIS3 y INV1 de Cándida utilis los cuales sirven como marcadores para la transformación en la cepa hospedera, así como para expresar proteínas heterólogas respectivamente en el caso de este último.The present invention relates to the field of biotechnology and recombinant DNA technology and in particular to a method for the reproducible genetic transformation of Candida utilis yeast, the recombinant DNA used for it and the obtained yeast strains. This invention is also related to new DNA sequences that could be used as promoters and terminators for protein expression in this yeast. Its technical objective is the development of a host-vector system that allows the expression and secretion of proteins of interest in this yeast. The present invention provides a method for stable transformation and maintenance of DNA in the host strain, where it is integrated into the genome by homologous recombination. The present invention provides a method for the expression of heterologous genes which uses as a host new auxotrophic mutants in yeast Candida utilis NRRL Y1084 which are defective in the orotidine-5'-phosphate decarboxylase enzyme of the biosynthetic pathway of uracil and an imidazole enzyme glycerol phosphate dehydratase from the biosynthetic pathway of histidine, respectively and were obtained by mutagenesis of a strain of the genus Candida. These mutants have high stability and are capable of being transformed with high efficiency by an integrative vector of the type used in this invention. Plasmids obtained from the library were used to complement mutations in Candida utilis. These plasmids contain the fragment associated with the URA3, HIS3 and INV1 genes of Candida utilis which serve as markers for transformation in the host strain, as well as to express heterologous proteins respectively in the case of the latter.
Priority Applications (9)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CU1996082A CU22722A1 (en) | 1996-10-03 | 1996-10-03 | TRANSFORMATION SYSTEM FOR THE EXPRESSION OF HETEROLOGICAL GENES IN CANDIDA UTILIS YEAST |
| CN 97199610 CN1237208A (en) | 1996-10-03 | 1997-10-03 | Method and material for candida utilis transformation |
| JP10516085A JP2001501475A (en) | 1996-10-03 | 1997-10-03 | Transformation system in Candida utilis |
| AU45485/97A AU744698B2 (en) | 1996-10-03 | 1997-10-03 | Candida utilis transformation system |
| CA002268004A CA2268004A1 (en) | 1996-10-03 | 1997-10-03 | Candida utilis transformation system |
| BR9713313-2A BR9713313A (en) | 1996-10-03 | 1997-10-03 | Candida utilis transformation system |
| RU99109095/13A RU2235127C2 (en) | 1996-10-03 | 1997-10-03 | Method for transformation of yeast candida utilis, yeast strain candida utilis (variants), recombinant dna material for transformation of yeast strain candida utilis, gene his3 encoding dna sequence in indicated yeast |
| EP97943729A EP0956356A1 (en) | 1996-10-03 | 1997-10-03 | $i(CANDIDA UTILIS) TRANSFORMATION SYSTEM |
| PCT/CU1997/000005 WO1998014600A1 (en) | 1996-10-03 | 1997-10-03 | Candida utilis transformation system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CU1996082A CU22722A1 (en) | 1996-10-03 | 1996-10-03 | TRANSFORMATION SYSTEM FOR THE EXPRESSION OF HETEROLOGICAL GENES IN CANDIDA UTILIS YEAST |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CU22722A1 true CU22722A1 (en) | 2002-02-28 |
Family
ID=5459318
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CU1996082A CU22722A1 (en) | 1996-10-03 | 1996-10-03 | TRANSFORMATION SYSTEM FOR THE EXPRESSION OF HETEROLOGICAL GENES IN CANDIDA UTILIS YEAST |
Country Status (9)
| Country | Link |
|---|---|
| EP (1) | EP0956356A1 (en) |
| JP (1) | JP2001501475A (en) |
| CN (1) | CN1237208A (en) |
| AU (1) | AU744698B2 (en) |
| BR (1) | BR9713313A (en) |
| CA (1) | CA2268004A1 (en) |
| CU (1) | CU22722A1 (en) |
| RU (1) | RU2235127C2 (en) |
| WO (1) | WO1998014600A1 (en) |
Families Citing this family (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003047471A (en) * | 2001-08-01 | 2003-02-18 | Kirin Brewery Co Ltd | Modified yeast excellent in secretory production of heterologous protein and method for producing heterologous protein using the yeast |
| MY142328A (en) * | 2002-03-26 | 2010-11-15 | Ajinomoto Kk | CANDIDA UTILIS CONTAINING y-GLUTAMYLCYSTEINE |
| JP4464269B2 (en) | 2002-04-26 | 2010-05-19 | 協和発酵キリン株式会社 | Methylotrophic yeast producing mammalian sugar chains |
| JP2005073638A (en) * | 2003-09-02 | 2005-03-24 | Ajinomoto Co Inc | Glutathione synthetase-coding gene of candida utilis |
| CN100347287C (en) * | 2004-09-30 | 2007-11-07 | 汪和睦 | Recombinated multi shape ttansenula yeast, its structural method and application |
| JP5954178B2 (en) | 2010-10-05 | 2016-07-20 | 味の素株式会社 | Yeast and yeast extract containing γ-Glu-Abu, and methods for producing them |
| EP3020801A4 (en) | 2013-07-12 | 2017-01-18 | Ajinomoto Co., Inc. | YEAST WITH HIGH CONTENT OF Abu, upsilon-Glu-Abu, AND/OR UPSILON-Glu-Abu-Gly |
| CN103468594B (en) * | 2013-07-31 | 2015-06-17 | 浙江科峰生物技术有限公司 | Candidautilis strain and application thereof |
| JPWO2022201917A1 (en) | 2021-03-22 | 2022-09-29 | ||
| EP4317461A4 (en) | 2021-03-29 | 2025-09-10 | Kaneka Corp | Cyclic lipopeptide-producing microbial strain and process for producing the cyclic lipopeptide |
| KR102556901B1 (en) * | 2021-05-25 | 2023-07-17 | 국립낙동강생물자원관 | Edenia sp. NNIBRFG15114 strain isolated from freshwater having antifungal activity and plant growth promotion and uses thereof |
| WO2023286629A1 (en) | 2021-07-16 | 2023-01-19 | 株式会社カネカ | Method for producing plasmid dna using escherichia coli |
| CN115838645B (en) * | 2022-09-15 | 2024-03-08 | 天津大学 | Yeast strain for high production of orotic acid and application thereof |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS59162884A (en) * | 1983-03-09 | 1984-09-13 | Kohjin Co Ltd | Novel hybrid plasmid and microorganism containing the same |
| NZ207925A (en) * | 1983-04-25 | 1988-05-30 | Genentech Inc | Yeast expression vehicle consisting of a yeast promoter and signal peptide encoding region linked to a heterologus peptide coding region; expression and culture |
| US5204252A (en) * | 1989-02-08 | 1993-04-20 | Henkel Research Corporation | Candida tropicalis transformation system |
| JPH08173170A (en) * | 1994-05-25 | 1996-07-09 | Kirin Brewery Co Ltd | Transformation system of Candida utilis yeast and expression of heterologous gene by it |
-
1996
- 1996-10-03 CU CU1996082A patent/CU22722A1/en not_active IP Right Cessation
-
1997
- 1997-10-03 CN CN 97199610 patent/CN1237208A/en active Pending
- 1997-10-03 BR BR9713313-2A patent/BR9713313A/en not_active IP Right Cessation
- 1997-10-03 JP JP10516085A patent/JP2001501475A/en not_active Ceased
- 1997-10-03 RU RU99109095/13A patent/RU2235127C2/en not_active IP Right Cessation
- 1997-10-03 WO PCT/CU1997/000005 patent/WO1998014600A1/en not_active Ceased
- 1997-10-03 CA CA002268004A patent/CA2268004A1/en not_active Abandoned
- 1997-10-03 EP EP97943729A patent/EP0956356A1/en not_active Withdrawn
- 1997-10-03 AU AU45485/97A patent/AU744698B2/en not_active Ceased
Also Published As
| Publication number | Publication date |
|---|---|
| EP0956356A1 (en) | 1999-11-17 |
| CA2268004A1 (en) | 1998-04-09 |
| AU4548597A (en) | 1998-04-24 |
| JP2001501475A (en) | 2001-02-06 |
| RU2235127C2 (en) | 2004-08-27 |
| AU744698B2 (en) | 2002-02-28 |
| CN1237208A (en) | 1999-12-01 |
| BR9713313A (en) | 2000-10-24 |
| WO1998014600A1 (en) | 1998-04-09 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| MK | Ceased due to expiration of term |