JP2000501617A - 酵母細胞におけるn末端を伸長されたタンパクの発現のためのベクター - Google Patents
酵母細胞におけるn末端を伸長されたタンパクの発現のためのベクターInfo
- Publication number
- JP2000501617A JP2000501617A JP9522315A JP52231597A JP2000501617A JP 2000501617 A JP2000501617 A JP 2000501617A JP 9522315 A JP9522315 A JP 9522315A JP 52231597 A JP52231597 A JP 52231597A JP 2000501617 A JP2000501617 A JP 2000501617A
- Authority
- JP
- Japan
- Prior art keywords
- insulin
- yeast
- dna
- heterologous protein
- peptide
- 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.)
- Granted
Links
Classifications
-
- 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
-
- 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
- C12N15/62—DNA sequences coding for fusion proteins
-
- 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
- C12N15/67—General methods for enhancing the expression
-
- 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
- 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
- C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
- C07K2319/74—Fusion polypeptide containing domain for protein-protein interaction containing a fusion for binding to a cell surface receptor
- C07K2319/75—Fusion polypeptide containing domain for protein-protein interaction containing a fusion for binding to a cell surface receptor containing a fusion for activation of a cell surface receptor, e.g. thrombopoeitin, NPY and other peptide hormones
Landscapes
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Mycology (AREA)
- Diabetes (AREA)
- Endocrinology (AREA)
- Toxicology (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Abstract
Description
Claims (1)
- 【特許請求の範囲】 1. ポリペプチドをコードし、且つ以下の構造[シグナルペプチド-リーダー ペプチド-EEAEPK-異種タンパク]を有するDNA構築物。 2. 請求項1に記載のDNA構築物であって、シグナルペプチドが酵母アスパラ ギン酸プロテアーゼ3(YAP3)シグナルペプチド又はその機能的類似物である構 築物。 3. 請求項1又は2のどちらか1項に記載のDNA構築物であって、前記リーダ ーペプチドが、合成リーダーペプチド、好ましくは本明細書中の配列認識番号5 のLA19リーダーペプチドである構築物。 4. 請求項1から3の何れか1項に記載のDNA構築物であって、異種タンパク が、以下の群[アプロチニン、ティッシュファクターパスウェイインヒビター( tisuue factor pathway inhibitor)又は他のプロテアーゼインヒビター、及び インスリン又はインスリン前駆体、インスリン類似物、インスリン様成長因子 I 若しくは II、ヒト成長ホルモン又はウシ成長ホルモン、インターロイキン、組 織型プラスミノーゲンアクチベーター、トランスフォーミング成長因子a若しく はb、グルカゴン、グルカゴン様ペプチド1(GLP-1)、グルカゴン様ペプチド2( GLP-2)、GRPP、第VII因子、第VIII因子、第XIII因子、血小板由来成長因子、酵 素(リパーゼ等)またはこれらのタンパクの何れか1つの機能的な類似物]から 選択される構築物。 5. 請求項1、2又は3の何れか1項に記載のDNA構築物であって、異種タン パクが、以下よりなる群[アプロチニン、ティッシュファクターパスウェイイン ヒビター又は他のプロテアーゼインヒビター、インスリン様成長因子I若しくは II、ヒト成長ホルモン若しくはウシ成長ホルモン、インターロイキン、組織型プ ラスミノーゲンアクチベーター、グルカゴン、グルカゴン様ペプチド1、第VII 因子、第VIII因子、第XIII因子、血小板成長因子、酵素、インスリン又はインス リン前駆体、及びこれら何れのタンパクの機能的な類似物]から選択される構築 物。 6. 請求項4又は5の何れか1項に記載のDNA構築物であり、異種タンパクが 、インスリン又はインスリン前駆体、好ましくは以下よりなる群[MI1、B(1-29) -A (1-21);MI3,B(1-29)-Ala-Ala-Lys-A(1-21);X14,B(1-27-Asp-Lys)-Ala-Ala-Lys-A (1-21);B(1-29)-Ala-Ala-Arg-A(1-21);MI5、B(1-29)-Ser-Asp-Asp-Ala-Lys-A(1 -21);及びB(1-29)-Ser-Asp-A(1-21)]から選択される構築物。 7. 請求項4又は5の何れか1項に記載のDNA構築物であって、異種タンパク が、インスリン又はインスリン前駆物質、好ましくはインスリン前駆体MI3、又 はその機能的な類似物である構築物。 8. 本明細書にある配列認識番号1、2、3又は4に記述のDNA構築物。 9. 真核生物細胞において複製が可能で、且つ請求項1から8の何れか1項に 記載のDNA構築物を有する組替え発現ベクター。 10.酵母において複製が可能、且つ請求項1から8の何れか1項に記載のDNA 構築物を有する組替え発現ベクター。 11.異種タンパクを発現することが可能であり、且つ請求項10に記載のベク ターを用いてトランスフォームされた酵母細胞。 12.サッカロミセス・セレビシアエ(Saccharomyces cerevisiae)、サッカロミ セス・クルイビリ(Saccharomyces Kluyveri)、シゾサッカロミセス・ポンベ(Sch izosaccharomyces pombe)、サッカロミセス・ウバルム(Saccharomyces uvarum) 、クルイベロミセス・ラクチス(Kluyveromyces lactis)、ハンセヌラ・ポリモル ファ(Hansenula polymorpha)、ピチア・パストリス(Pichia pastoris)、ピチア ・メタノリカ(Pichia methanolica)、ピチア・クルイベリ(Pichia Kluyveri)、 ヤロウイア・リポリチカ(Yarrowia lipolytica)、カンジダ・ sp.(Candida sp.) 、カンジダ・ウチリス(Candida utilis)、カンジダ・カカオイ(Candida cacaoi) 、ゲオトリクウム・ sp.(Geotrichum sp.)及びゲオトリクウム・フェロメンタン ス(Geotrichum fermentans)の種類から選択された請求項11に記載の酵母細 胞であり、好ましくはサッカロミセス・セレビシアエである酵母細胞。 13.異種タンパクを産生するための方法であって、請求項11又は12の何れ か1項に記載の酵母細胞を適切な培地中で培養して、異種タンパクを発現及び分 泌させることと、その後、前記タンパクを培養用培地及び/又は前記細胞から単 離することとを具備した方法。 14.異種タンパクを産生するための方法であって、請求項11又は12の何 れか1項に記載の酵母細胞を適切な培地中で培養して、異種タンパクを発現及び 分泌させることと、その後、前記タンパクを培養用培地から単離することとを具 備した方法。 15.請求項13又は14の何れか1項に記載の方法であって、蛋白分解酵素( 塩基性アミノ酸に特異的)で処理を伴う方法により回収した異種タンパクから、 アミノ酸配列EEAEPKを取り除く方法。 16.請求項15に記載の方法であって、前記蛋白分解酵素がトリプシンとアク ロモバクテロ・リチクス・プロテアーゼI(Achromobacter lyticus protease I )とからなる群より選択される方法。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DK144995 | 1995-12-20 | ||
DK1449/95 | 1995-12-20 | ||
PCT/DK1996/000542 WO1997022706A1 (en) | 1995-12-20 | 1996-12-18 | Vector for expression of n-terminally extended proteins in yeast cell |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2000501617A true JP2000501617A (ja) | 2000-02-15 |
JP4180112B2 JP4180112B2 (ja) | 2008-11-12 |
Family
ID=8104944
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP52231597A Expired - Lifetime JP4180112B2 (ja) | 1995-12-20 | 1996-12-18 | 酵母細胞におけるn末端を伸長されたタンパクの発現のためのベクター |
Country Status (11)
Country | Link |
---|---|
EP (1) | EP0868523B1 (ja) |
JP (1) | JP4180112B2 (ja) |
CN (1) | CN1125181C (ja) |
AT (1) | ATE295890T1 (ja) |
AU (1) | AU1093597A (ja) |
CA (1) | CA2241367C (ja) |
DE (1) | DE69634757T2 (ja) |
ES (1) | ES2242969T3 (ja) |
IL (1) | IL124939A (ja) |
WO (1) | WO1997022706A1 (ja) |
ZA (1) | ZA9610456B (ja) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7595172B2 (en) | 2001-07-24 | 2009-09-29 | Novo Nordisk A/S | Method for making acylated polypeptides |
Families Citing this family (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
ES2306462T3 (es) * | 1996-12-13 | 2008-11-01 | Novartis Vaccines And Diagnostics, Inc. | Analisis y separacion de proteinas del factor de crecimiento derivadas de plaquetas. |
AU9162398A (en) * | 1997-08-22 | 1999-03-16 | Boehringer Mannheim Gmbh | Process for producing insulin precursor proteins |
PT2270030E (pt) * | 2000-02-28 | 2012-07-24 | Novartis Vaccines & Diagnostic | Expressão heteróloga de proteínas de neisseria |
EP1381671A2 (en) * | 2001-04-24 | 2004-01-21 | Innogenetics N.V. | Constructs and methods for expression of recombinant hcv envelope proteins |
WO2005121333A1 (en) * | 2004-06-14 | 2005-12-22 | Novozymes A/S | Signal peptide for producing a polypeptide |
GB201008682D0 (en) | 2010-05-25 | 2010-07-07 | Vib Vzw | Epitope tag for affinity based applications |
CN111560379B (zh) * | 2020-05-26 | 2022-09-13 | 北京惠之衡生物科技有限公司 | 密码子优化的门冬胰岛素前体基因、重组载体、基因工程菌及其应用 |
CN113249241B (zh) * | 2021-06-21 | 2021-10-15 | 奥锐特药业(天津)有限公司 | 一种酿酒酵母蛋白酶缺失菌株的构建及其应用 |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0116201B1 (en) * | 1983-01-12 | 1992-04-22 | Chiron Corporation | Secretory expression in eukaryotes |
FR2593518B1 (fr) * | 1985-05-02 | 1989-09-08 | Transgene Sa | Vecteurs d'expression et de secretion de l'hirudine par les levures transformees |
CA1340772C (en) * | 1987-12-30 | 1999-09-28 | Patricia Tekamp-Olson | Expression and secretion of heterologous protiens in yeast employing truncated alpha-factor leader sequences |
DK105489D0 (da) * | 1989-03-03 | 1989-03-03 | Novo Nordisk As | Polypeptid |
DK82893D0 (da) * | 1993-07-08 | 1993-07-08 | Novo Nordisk As | Peptid |
US5639642A (en) * | 1994-06-16 | 1997-06-17 | Novo Nordisk A/S | Synthetic leader peptide sequences |
ZA954983B (en) * | 1994-06-17 | 1996-02-14 | Novo Nordisk As | N-terminally extended proteins expressed in yeast |
-
1996
- 1996-12-12 ZA ZA9610456A patent/ZA9610456B/xx unknown
- 1996-12-18 AT AT96941597T patent/ATE295890T1/de not_active IP Right Cessation
- 1996-12-18 WO PCT/DK1996/000542 patent/WO1997022706A1/en active IP Right Grant
- 1996-12-18 CA CA002241367A patent/CA2241367C/en not_active Expired - Fee Related
- 1996-12-18 AU AU10935/97A patent/AU1093597A/en not_active Abandoned
- 1996-12-18 JP JP52231597A patent/JP4180112B2/ja not_active Expired - Lifetime
- 1996-12-18 ES ES96941597T patent/ES2242969T3/es not_active Expired - Lifetime
- 1996-12-18 CN CN96199732A patent/CN1125181C/zh not_active Expired - Lifetime
- 1996-12-18 IL IL124939A patent/IL124939A/en not_active IP Right Cessation
- 1996-12-18 EP EP96941597A patent/EP0868523B1/en not_active Expired - Lifetime
- 1996-12-18 DE DE69634757T patent/DE69634757T2/de not_active Expired - Lifetime
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7595172B2 (en) | 2001-07-24 | 2009-09-29 | Novo Nordisk A/S | Method for making acylated polypeptides |
US8835132B2 (en) | 2001-07-24 | 2014-09-16 | Novo Nordisk A/S | Method for making acylated polypeptides |
Also Published As
Publication number | Publication date |
---|---|
IL124939A0 (en) | 1999-01-26 |
DE69634757T2 (de) | 2006-02-02 |
ES2242969T3 (es) | 2005-11-16 |
CN1207773A (zh) | 1999-02-10 |
EP0868523B1 (en) | 2005-05-18 |
WO1997022706A1 (en) | 1997-06-26 |
CA2241367C (en) | 2007-02-06 |
DE69634757D1 (de) | 2005-06-23 |
AU1093597A (en) | 1997-07-14 |
ATE295890T1 (de) | 2005-06-15 |
CN1125181C (zh) | 2003-10-22 |
IL124939A (en) | 2007-06-03 |
CA2241367A1 (en) | 1997-06-26 |
JP4180112B2 (ja) | 2008-11-12 |
ZA9610456B (en) | 1997-06-20 |
EP0868523A1 (en) | 1998-10-07 |
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