ES2180528T3 - Metodo de expresion de genes heterologos en la levadura pichia pastoris, vectores de expresion y microorganismos transformados. - Google Patents

Metodo de expresion de genes heterologos en la levadura pichia pastoris, vectores de expresion y microorganismos transformados.

Info

Publication number
ES2180528T3
ES2180528T3 ES91200074T ES91200074T ES2180528T3 ES 2180528 T3 ES2180528 T3 ES 2180528T3 ES 91200074 T ES91200074 T ES 91200074T ES 91200074 T ES91200074 T ES 91200074T ES 2180528 T3 ES2180528 T3 ES 2180528T3
Authority
ES
Spain
Prior art keywords
expression
pichia pastoris
cerevisiae
gene
expression vectors
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.)
Expired - Lifetime
Application number
ES91200074T
Other languages
English (en)
Inventor
Martinez Luis Saturnin Herrera
Gonzalez Vladimir Yong
Clark Emilio Margollez
Boada Julio Marcos Delgado
Grillo Juan Morales
Madrazo Isis Del Carme Torrens
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Centro de Ingenieria Genetica y Biotecnologia CIGB
Original Assignee
Centro de Ingenieria Genetica y Biotecnologia CIGB
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
Priority claimed from CU1990007A external-priority patent/CU22276A1/es
Priority claimed from CU1990104A external-priority patent/CU22278A1/es
Priority claimed from CU1990132A external-priority patent/CU22287A1/es
Priority claimed from CU1990142A external-priority patent/CU22288A1/es
Application filed by Centro de Ingenieria Genetica y Biotecnologia CIGB filed Critical Centro de Ingenieria Genetica y Biotecnologia CIGB
Application granted granted Critical
Publication of ES2180528T3 publication Critical patent/ES2180528T3/es
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

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Classifications

    • 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
    • C12N9/2411—Amylases
    • C12N9/2414—Alpha-amylase (3.2.1.1.)
    • C12N9/2417—Alpha-amylase (3.2.1.1.) from microbiological source
    • 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
    • C07K14/475—Growth factors; Growth regulators
    • C07K14/485—Epidermal growth factor [EGF], i.e. urogastrone
    • 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
    • C12N15/815—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts for yeasts other than Saccharomyces
    • 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)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
    • C12N9/6478—Aspartic endopeptidases (3.4.23)
    • C12N9/6481—Pepsins (3.4.23.1; 3.4.23.2; 3.4.23.3)
    • 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)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
    • C12N9/6478—Aspartic endopeptidases (3.4.23)
    • C12N9/6483—Chymosin (3.4.23.4), i.e. rennin
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
    • C12Y304/23—Aspartic endopeptidases (3.4.23)
    • C12Y304/23004—Chymosin (3.4.23.4), i.e. rennin
    • 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

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Microbiology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Biophysics (AREA)
  • Mycology (AREA)
  • Toxicology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

LA PRESENTE INVENCION PROPORCIONA UN METODO ALTAMENTE EFICAZ PARA LA EXPRESION DE GENES HETEROLOGOS, USANDO COMO HUESPED UN AUXOTROFICO MUTANTE HIS-3 DE LA CEPA BKM-90 DE "PICHIA PASTORIS" Y COMO ADN RECOMBINANTE PARA LA TRANSFORMACION DEL MISMO, UN VECTOR INTEGRADOR QUE CONTIENE UN MARCADOR DE SELECCION FUNCIONAL DE "SACAROMICES CEREVISIAE", PROMOTOR DEL GEN ALCOHOL OXIDASA (AOX SUB 1) DE LA PROPIA LEVADURA, COMO DETERMINADOR DE LA TRANSCRIPCION EL DE LA GLICERALDEHIDO 3-FOSFATO DESHIDROGENASA (GAP) DE "S. CEREVISIAE", O EL QUE LLEVA EL GEN HETEROLOGO QUE VAMOS A EXPRESAR, Y OTRA SECUENCIA HOMOLOGA AL GENOMA DE "PICHIA PASTORIS", QUE SIRVE PARA LA INTEGRACION DEL MISMO. LOS VECTORES DE EXPRESION USADOS POSTERIORMENTE CONTIENEN UN GEN HETEROLOGO UNIDO AL PEPTIDO INDICADO DEL GEN DE LA "SUCROSA INVERTASA" (SUC2) DE "S. CEREVISIAE". ESTE METODO GARANTIZA QUE LOS PRODUCTOS DE POLIPEPTIDOS SON RECUPERABLES EN CONDICIONES CONTROLABLES Y CON ALTOS RENDIMIENTOS. LAS CEPAS DE LA PRESENTE INVENCIONPUEDEN SER UTILIZADAS PARA LA PRODUCCION A GRAN ESCALA DE PRODUCTOS FARMACEUTICOS O DE ENZIMAS DE INTERES INDUSTRIAL.
ES91200074T 1990-01-16 1991-01-16 Metodo de expresion de genes heterologos en la levadura pichia pastoris, vectores de expresion y microorganismos transformados. Expired - Lifetime ES2180528T3 (es)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
CU1990007A CU22276A1 (es) 1990-01-16 1990-01-16 Procedimiento para la expresion de genes heterologos en la levadura pichia pastoris, vectores de expresion y microorganismos transformados
CU1990104A CU22278A1 (es) 1990-06-13 1990-06-13 Cepa de levadura recombinante productora de una proteasa aspartica con actividad coagulante de la leche y procedimiento para su obtencion
CU1990132A CU22287A1 (es) 1990-07-12 1990-07-12 Cepa de levadura recombinante productora de quimosina bovina extracelular y procedimiento de obtencion
CU1990142A CU22288A1 (es) 1990-08-15 1990-08-15 Levadura recombinante hiperprotectora de enzima alfa amilasa termoestable y procedimiento para su obtencion

Publications (1)

Publication Number Publication Date
ES2180528T3 true ES2180528T3 (es) 2003-02-16

Family

ID=27430148

Family Applications (1)

Application Number Title Priority Date Filing Date
ES91200074T Expired - Lifetime ES2180528T3 (es) 1990-01-16 1991-01-16 Metodo de expresion de genes heterologos en la levadura pichia pastoris, vectores de expresion y microorganismos transformados.

Country Status (6)

Country Link
EP (1) EP0438200B1 (es)
JP (1) JPH05184352A (es)
AT (1) ATE220721T1 (es)
DE (1) DE69133066T2 (es)
DK (1) DK0438200T3 (es)
ES (1) ES2180528T3 (es)

Families Citing this family (24)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CU22292A1 (es) * 1991-05-07 1995-01-31 Cigb Procedimiento para la obtencion a escala industrial de licores de fructosa-glucosa a partir de sacarosa e instalacion para el mismo
JP3113435B2 (ja) * 1992-02-28 2000-11-27 サントリー株式会社 メタノールおよび／またはグリセロール添加により誘導可能なプロモーターを有する新規ベクター
GB9402832D0 (en) * 1994-02-15 1994-04-06 Smith Kline Beecham Biolog S A Novel process
CA2211417A1 (en) * 1995-01-30 1996-08-08 New York Blood Center, Inc. Recombinant .alpha.-galactosidase enzyme
SE9501939D0 (sv) * 1995-05-24 1995-05-24 Astra Ab DNA molecules for expression of polypeptides
EP0852625A2 (en) * 1995-09-25 1998-07-15 Chiron Corporation Pichia secretory leader for protein expression
CA2203613C (en) * 1997-04-24 2008-01-15 Navneet K. Ahluwalia Secreted .alpha.-amylase as a reporter gene
FR2765589B1 (fr) * 1997-07-03 1999-09-17 Roquette Freres Fragment d'adn comprenant le promoteur de la ribulose reductase nadph specifique de yamadazyma ohmeri, vecteur le contenant et procede de preparation de proteines utilisant ce vecteur
US6358705B1 (en) 1998-07-16 2002-03-19 Novo Nordisk A/S Method of making proteins in transformed yeast cells
WO2000078978A1 (en) * 1999-06-24 2000-12-28 Zymogenetics, Inc. Pichia methanolica glyceraldehyde 3-phosphate dehydrogenase 1 promoter and terminator
US6440414B1 (en) 1999-10-01 2002-08-27 Amgen Inc. Pharmaceutical compositions of fibrinolytic agent
US6261820B1 (en) 1999-10-01 2001-07-17 Amgen Inc. Fibronolytically active polypeptide
US7033776B2 (en) 1999-12-17 2006-04-25 Amgen Inc. Method for treatment of indwelling catheter occlusion using fibrinolytic metalloproteinases
KR100365533B1 (ko) * 2000-05-10 2002-12-18 재단법인 목암생명공학연구소 상처치유 효과를 가진 재조합 거머린의 제조방법 및 이를 함유하는 약학적 조성물
JP2008507266A (ja) * 2004-07-22 2008-03-13 グライコフィ， インコーポレイテッド ピチアにおける異種遺伝子発現のためのリンゴ酸シンターゼ調節配列
CU23577A1 (es) 2005-07-21 2010-09-30 Ct Ingenieria Genetica Biotech Polipéptidos estimuladores del crecimiento para uso en peces y crustáceos
CU23636A1 (es) 2006-11-01 2011-03-21 Ct Ingenieria Genetica Biotech Anticuerpos recombinantes contra el factor de crecimiento del endotelio vascular (vegf)
CU23634A1 (es) 2007-05-31 2011-02-24 Ct Ingenieria Genetica Biotech Secuencias de ácido nucleico y aminoácidos, y vacuna para el control de infestaciones por ectoparásitos en peces
ES2336171B1 (es) * 2007-07-27 2011-05-11 Universidad De Santiago De Compostela Uso de la quimosina recombinante activa.
EP2258855A1 (en) 2009-05-28 2010-12-08 Universität für Bodenkultur Wien Expression sequences
KR102079293B1 (ko) 2012-10-29 2020-02-19 론자 리미티드 발현 서열
CN103882050A (zh) * 2014-01-02 2014-06-25 杭州璞题生物科技有限公司 鼠表皮生长因子在巴斯德毕赤酵母中的表达方法和纯化
CN110894219A (zh) * 2019-12-26 2020-03-20 河北省微生物研究所 毕赤酵母转录因子hac1、蛋白、巴斯德毕赤酵母及其制备和应用
WO2026009000A1 (en) 2024-07-04 2026-01-08 Solasta Bio Limited Methods for production of insecticidal neuropeptides

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1985000369A1 (en) * 1983-07-05 1985-01-31 Chiron Corporation Hybrid dna synthesis of epidermal growth factor
US4721673A (en) * 1983-09-01 1988-01-26 Genex Corporation Recovery and activation process for microbially produced calf prochymosin
US5002876A (en) * 1986-09-22 1991-03-26 Phillips Petroleum Company Yeast production of human tumor necrosis factor
IL83435A (en) * 1986-09-23 1993-01-14 Miles Lab Dna construct for expression of genes in yeast, an expression vector containing it and yeast transformed with said expression vector
US4997757A (en) * 1987-12-23 1991-03-05 Genebiomed, Inc. Process for detecting potential carcinogens
JPH01240191A (ja) * 1988-02-16 1989-09-25 Green Cross Corp:The 酵母で機能する新規シグナルペプチドおよびこれを用いた異種蛋白質の分泌発現

Also Published As

Publication number Publication date
EP0438200B1 (en) 2002-07-17
ATE220721T1 (de) 2002-08-15
DK0438200T3 (da) 2002-11-11
DE69133066D1 (de) 2002-08-22
EP0438200A1 (en) 1991-07-24
JPH05184352A (ja) 1993-07-27
DE69133066T2 (de) 2003-03-06

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