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 YEAST

Info

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
Application number
CU1996082A
Other languages
Spanish (es)
Inventor
Tuero Liliana Basabe
Menocal Luis Rodriguez
Baeza Tanilo Rivero
Espinosa Francisco Pablo Chavez
Martinez Maria Elena Gonzalez
Reyes Edenia Paifer
Boada Julio Marcos Delgado
Original Assignee
Ct Ingenieria Genetica Biotech
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Ct Ingenieria Genetica Biotech filed Critical Ct Ingenieria Genetica Biotech
Priority to CU1996082A priority Critical patent/CU22722A1/en
Priority to CN 97199610 priority patent/CN1237208A/en
Priority to JP10516085A priority patent/JP2001501475A/en
Priority to AU45485/97A priority patent/AU744698B2/en
Priority to RU99109095/13A priority patent/RU2235127C2/en
Priority to PCT/CU1997/000005 priority patent/WO1998014600A1/en
Priority to BR9713313-2A priority patent/BR9713313A/en
Priority to EP97943729A priority patent/EP0956356A1/en
Priority to CA002268004A priority patent/CA2268004A1/en
Publication of CU22722A1 publication Critical patent/CU22722A1/en

Links

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)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Biomedical Technology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Mycology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Plant Pathology (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.

CU1996082A 1996-10-03 1996-10-03 TRANSFORMATION SYSTEM FOR THE EXPRESSION OF HETEROLOGICAL GENES IN CANDIDA UTILIS YEAST CU22722A1 (en)

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
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
PCT/CU1997/000005 WO1998014600A1 (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
EP97943729A EP0956356A1 (en) 1996-10-03 1997-10-03 $i(CANDIDA UTILIS) TRANSFORMATION SYSTEM
CA002268004A CA2268004A1 (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)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2003047471A (en) * 2001-08-01 2003-02-18 Kirin Brewery Co Ltd Modified yeast having excellent secretory production of foreign protein and method for producing foreign protein by using the yeast
MY142328A (en) 2002-03-26 2010-11-15 Ajinomoto Kk CANDIDA UTILIS CONTAINING y-GLUTAMYLCYSTEINE
ATE420198T1 (en) 2002-04-26 2009-01-15 Kirin Pharma Kk METHYLOTROPHIC YEAST THAT PRODUCES A MAMMAL SUGAR CHAIN
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
EP2626430B1 (en) 2010-10-05 2017-07-12 Ajinomoto Co., Inc. Yeast and yeast extract containing gamma-glu-abu, and method for producing the yeast or yeast extract containing gamma-glu-abu
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
EP4317169A1 (en) 2021-03-22 2024-02-07 Kaneka Corporation Production method for foreign protein using escherichia coli
CN117203327A (en) 2021-03-29 2023-12-08 株式会社钟化 Cyclic lipopeptide-producing microorganism strain and method for producing 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)

* Cited by examiner, † Cited by third party
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 therewith

Also Published As

Publication number Publication date
AU744698B2 (en) 2002-02-28
JP2001501475A (en) 2001-02-06
AU4548597A (en) 1998-04-24
RU2235127C2 (en) 2004-08-27
CA2268004A1 (en) 1998-04-09
CN1237208A (en) 1999-12-01
EP0956356A1 (en) 1999-11-17
BR9713313A (en) 2000-10-24
WO1998014600A1 (en) 1998-04-09

Similar Documents

Publication Publication Date Title
Chatton et al. The yeast VAS1 gene encodes both mitochondrial and cytoplasmic valyl-tRNA synthetases.
CU22722A1 (en) TRANSFORMATION SYSTEM FOR THE EXPRESSION OF HETEROLOGICAL GENES IN CANDIDA UTILIS YEAST
Allison et al. Artificial mitochondrial presequences.
CN105695485B (en) A Cas9-encoding gene for filamentous fungal Crispr-Cas system and its application
Zhang et al. Application of the CRISPR/Cas system for genome editing in microalgae
Nuttley et al. Rapid identification and characterization of peroxisomal assembly mutants in Yarrowia lipolytica
Remacha et al. Ribosomal acidic phosphoproteins P1 and P2 are not required for cell viability but regulate the pattern of protein expression in Saccharomyces cerevisiae
Hurt et al. A cytosolic protein contains a cryptic mitochondrial targeting signal
WO2002014495B1 (en) Enhanced homologous recombination mediated by lambda recombination proteins
ES2902398T3 (en) Pichia pastoris yeast promoters
MX9307623A (en) PROTEIN PRODUCTION AND PROTEIN ADMINISTRATION.
KR102345759B1 (en) Methods for modulating biosynthetic activity in vitro by knock-out of nuclease systems
JP2020528760A (en) Use in the preparation of novel fusion proteins and the improvement of their protein synthesis
Cloney et al. Co-expression of plastid chaperonin genes and a synthetic plant Rubisco operon in Escherichia coli
Wang et al. Use of CRISPR‐Cas tools to engineer Trichoderma species
Nomaguchi et al. Comprehensive analysis of triacylglycerol lipases in the oleaginous diatom Fistulifera solaris JPCC DA0580 with transcriptomics under lipid degradation
DK171879B1 (en) Method of Transforming Yarrowia Lipolytica and Method of Cloning Genes by Complementing Mutants in Y.lipolytica
Bai et al. Establishment of genomic library technology mediated by non-homologous end joining mechanism in Yarrowia lipolytica
PT86684B (en) PROCESS FOR THE PREPARATION OF NEW EXPRESSION SYSTEMS
DE69938722D1 (en) METHOD FOR THE PRODUCTION OF HETEROLOGIC POLYPEPTIDES IN TRICHOTHECENE LACK OF FILAMENT SOLVENTS MUSHROOMS
Morin et al. Transforming Candida hispaniensis, a promising oleaginous and flavogenic yeast
ES2719106T3 (en) Rhodosporidium and Rhodotorula polynucleotide sequences and their uses
Numata et al. Direct introduction of neomycin phosphotransferase II protein into apple leaves to confer kanamycin resistance
EP0045573A2 (en) Eukaryotic autonomously replicating segment
Marten et al. Localization of cloned invertase in Saccharomyces cerevisiae directed by the SUC2 and MFα1 signal sequences

Legal Events

Date Code Title Description
MK Ceased due to expiration of term