CA2849856C - Regulatable promoter - Google Patents
Regulatable promoter Download PDFInfo
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- CA2849856C CA2849856C CA2849856A CA2849856A CA2849856C CA 2849856 C CA2849856 C CA 2849856C CA 2849856 A CA2849856 A CA 2849856A CA 2849856 A CA2849856 A CA 2849856A CA 2849856 C CA2849856 C CA 2849856C
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Classifications
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- 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
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- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
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- 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
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- 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
- C12N15/635—Externally inducible repressor mediated regulation of gene expression, e.g. tetR inducible by tetracyline
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0008—Oxidoreductases (1.) acting on the aldehyde or oxo group of donors (1.2)
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- C12Y102/00—Oxidoreductases acting on the aldehyde or oxo group of donors (1.2)
- C12Y102/01—Oxidoreductases acting on the aldehyde or oxo group of donors (1.2) with NAD+ or NADP+ as acceptor (1.2.1)
- C12Y102/01012—Glyceraldehyde-3-phosphate dehydrogenase (phosphorylating) (1.2.1.12)
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- C07K2317/00—Immunoglobulins specific features
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- C07K2317/55—Fab or Fab'
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
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- Gastroenterology & Hepatology (AREA)
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- General Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
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Applications Claiming Priority (7)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201161544451P | 2011-10-07 | 2011-10-07 | |
| EP11184323 | 2011-10-07 | ||
| US61/544,451 | 2011-10-07 | ||
| EP11184323.1 | 2011-10-07 | ||
| EP12171006.5 | 2012-06-06 | ||
| EP12171006 | 2012-06-06 | ||
| PCT/EP2012/069757 WO2013050551A1 (en) | 2011-10-07 | 2012-10-05 | Regulatable promoter |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2849856A1 CA2849856A1 (en) | 2013-04-11 |
| CA2849856C true CA2849856C (en) | 2023-03-14 |
Family
ID=48043194
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2849856A Active CA2849856C (en) | 2011-10-07 | 2012-10-05 | Regulatable promoter |
Country Status (17)
| Country | Link |
|---|---|
| US (4) | US9512432B2 (en, 2012) |
| EP (2) | EP3508494B1 (en, 2012) |
| JP (1) | JP6113736B2 (en, 2012) |
| KR (2) | KR101954389B1 (en, 2012) |
| CN (3) | CN103998460A (en, 2012) |
| AR (1) | AR088252A1 (en, 2012) |
| AU (1) | AU2012320421B2 (en, 2012) |
| BR (1) | BR112014008152A2 (en, 2012) |
| CA (1) | CA2849856C (en, 2012) |
| DK (1) | DK3508494T3 (en, 2012) |
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| PL (1) | PL2764016T3 (en, 2012) |
| SG (2) | SG11201400811QA (en, 2012) |
| TW (1) | TWI560272B (en, 2012) |
| WO (1) | WO2013050551A1 (en, 2012) |
Families Citing this family (23)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103998460A (zh) * | 2011-10-07 | 2014-08-20 | 隆扎有限公司 | 可调控启动子 |
| CN104195165A (zh) * | 2014-09-01 | 2014-12-10 | 北京爱必信生物技术有限公司 | 包含葡萄糖启动子的载体、宿主菌以及重组牛源胰蛋白酶的制备方法 |
| WO2016077457A1 (en) | 2014-11-11 | 2016-05-19 | Clara Foods Co. | Methods and compositions for egg white protein production |
| CN108350447B (zh) * | 2015-08-05 | 2021-08-31 | 龙沙有限公司 | 启动子变体 |
| IL261411B2 (en) | 2016-03-02 | 2024-03-01 | Lonza Ag | Improved fermentation process |
| WO2018110616A1 (ja) * | 2016-12-15 | 2018-06-21 | 株式会社カネカ | 新規宿主細胞及びそれを用いた目的タンパク質の製造方法 |
| SG11202006449RA (en) | 2018-02-12 | 2020-08-28 | Lonza Ag | Host cell for producing a protein of interest |
| EP3536784A1 (en) | 2018-03-05 | 2019-09-11 | ACIB GmbH | Host cell engineered for improved metabolite production |
| EP3802832A1 (en) * | 2018-06-07 | 2021-04-14 | Basf Se | Promoter for yeast |
| KR102495283B1 (ko) | 2019-01-11 | 2023-02-06 | 론자 리미티드 | 재조합 숙주 세포에서 탄소 공급원 조절된 단백질 생산 |
| HUE062297T2 (hu) * | 2019-01-29 | 2023-10-28 | Clariant Produkte Deutschland Gmbh | Kiméra promóter anyagcsere módosítási alkalmazásra |
| US12270034B2 (en) | 2019-04-01 | 2025-04-08 | Universität Für Bodenkultur Wien | Mut- methylotrophic yeast |
| WO2020200414A1 (en) | 2019-04-01 | 2020-10-08 | Lonza Ltd | Protein production in mut-methylotrophic yeast |
| WO2020200415A1 (en) | 2019-04-01 | 2020-10-08 | Lonza Ltd | Mut- methylotrophic yeast |
| US12096784B2 (en) | 2019-07-11 | 2024-09-24 | Clara Foods Co. | Protein compositions and consumable products thereof |
| AU2020309602B2 (en) | 2019-07-11 | 2025-05-15 | Clara Foods Co. | Protein compositions and consumable products thereof |
| US10927360B1 (en) | 2019-08-07 | 2021-02-23 | Clara Foods Co. | Compositions comprising digestive enzymes |
| EP4127180A1 (en) | 2020-03-26 | 2023-02-08 | Asklepios Biopharmaceutical, Inc. | Forskolin-inducible promoters and hypoxia-inducible promoters |
| ES3034258T3 (en) | 2020-04-01 | 2025-08-14 | Lonza Ag | Helper factors for expressing proteins in yeast |
| US20230167458A1 (en) | 2020-04-15 | 2023-06-01 | Asklepios Biopharmaceutical, Inc. | Forskolin-inducible promoters and hypoxia-inducible promoters |
| CN116490517A (zh) | 2020-09-30 | 2023-07-25 | 龙沙有限公司 | 过表达翻译因子的宿主细胞 |
| KR20230144629A (ko) | 2021-02-12 | 2023-10-16 | 베링거 인겔하임 에르체파우 게엠베하 운트 코 카게 | 단백질 분비 증가를 위한 신호 펩타이드 |
| WO2024133538A1 (en) | 2022-12-20 | 2024-06-27 | Lonza Ltd | Host cells with pat1 knockout for increased specific protein productivity |
Family Cites Families (20)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU577259B2 (en) | 1982-08-13 | 1988-09-22 | Zymogenetics Inc. | Glycolytic promters for regulated protein expression protease inhibitor |
| US4855231A (en) | 1984-10-30 | 1989-08-08 | Phillips Petroleum Company | Regulatory region for heterologous gene expression in yeast |
| US4808537A (en) | 1984-10-30 | 1989-02-28 | Phillips Petroleum Company | Methanol inducible genes obtained from pichia and methods of use |
| AU2887897A (en) | 1996-05-21 | 1997-12-09 | Novo Nordisk A/S | Novel yeast promoters suitable for expression cloning in yeast and heterologous expression of proteins in yeast |
| SE9802333D0 (sv) | 1998-06-29 | 1998-06-29 | Astra Pharma Prod | Novel combination |
| US6730499B1 (en) | 1998-07-03 | 2004-05-04 | Research Corporation Technologies, Inc. | Promoter for the Pichia pastoris formaldehyde dehydrogenase gene FLD1 |
| US7259255B2 (en) | 2003-06-25 | 2007-08-21 | E. I. Du Pont De Nemours And Company | Glyceraldehyde-3-phosphate dehydrogenase and phosphoglycerate mutase promoters for gene expression in oleaginous yeast |
| 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 |
| AT501955B1 (de) | 2005-02-23 | 2007-08-15 | Univ Graz Tech | Mutierte aox1-promotoren |
| WO2007058407A1 (en) | 2005-11-16 | 2007-05-24 | Korea Research Institute Of Bioscience & Biotechnology | Translational elongation factor promoter from pichia pastoris and method for producing recombinant protein using the same |
| KR100717353B1 (ko) | 2006-04-12 | 2007-05-11 | 한국생명공학연구원 | 피키아 파스토리스 유래의 자동 유도성 ｎｐｓ 프로모터 및이를 이용한 이종 단백질의 제조방법 |
| CN100552029C (zh) | 2006-09-14 | 2009-10-21 | 南开大学 | 毕赤酵母ω3-脂肪酸脱氢酶启动子序列及其应用 |
| US8222386B2 (en) | 2006-10-10 | 2012-07-17 | Keck Graduate Institute | P. pastoris ADH promoter and use thereof to direct expression of proteins |
| KR20100016170A (ko) | 2007-04-20 | 2010-02-12 | 폴리문 사이언티픽 임무노이비오로기쉐 포르슝 게엠베하 | 이스트 발현 시스템 |
| US8105811B2 (en) | 2008-06-13 | 2012-01-31 | Wisconsin Alumni Research Foundation | Sugar transport sequences, yeast strains having improved sugar uptake, and methods of use |
| WO2010135678A1 (en) | 2009-05-22 | 2010-11-25 | Research Corporation Technologies, Inc. | Nucleic acids of pichia pastoris and use thereof for recombinant production of proteins |
| EP2451960A2 (en) * | 2009-07-09 | 2012-05-16 | Verdezyne, Inc. | Engineered microorganisms with enhanced fermentation activity |
| EP2292769A1 (en) * | 2009-08-10 | 2011-03-09 | Lonza AG | Fermentation process |
| CN103998460A (zh) * | 2011-10-07 | 2014-08-20 | 隆扎有限公司 | 可调控启动子 |
| CN108350447B (zh) | 2015-08-05 | 2021-08-31 | 龙沙有限公司 | 启动子变体 |
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