MX2014009118A - Metodo de expresion. - Google Patents
Metodo de expresion.Info
- Publication number
- MX2014009118A MX2014009118A MX2014009118A MX2014009118A MX2014009118A MX 2014009118 A MX2014009118 A MX 2014009118A MX 2014009118 A MX2014009118 A MX 2014009118A MX 2014009118 A MX2014009118 A MX 2014009118A MX 2014009118 A MX2014009118 A MX 2014009118A
- Authority
- MX
- Mexico
- Prior art keywords
- protein
- expression construct
- aim
- expression method
- microorganism
- Prior art date
Links
- 238000000034 method Methods 0.000 title abstract 2
- 102000004169 proteins and genes Human genes 0.000 abstract 3
- 108090000623 proteins and genes Proteins 0.000 abstract 3
- 241000194103 Bacillus pumilus Species 0.000 abstract 1
- 238000000855 fermentation Methods 0.000 abstract 1
- 230000004151 fermentation Effects 0.000 abstract 1
- 230000000813 microbial effect Effects 0.000 abstract 1
- 244000005700 microbiome Species 0.000 abstract 1
- 102000039446 nucleic acids Human genes 0.000 abstract 1
- 108020004707 nucleic acids Proteins 0.000 abstract 1
- 150000007523 nucleic acids Chemical class 0.000 abstract 1
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
-
- 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
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- 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
- C12N15/74—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
- C12N15/75—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora for Bacillus
-
- 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
-
- 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.)
-
- 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)
-
- 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)
- C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
- C12N9/54—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea bacteria being Bacillus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
-
- 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
- C12N2330/00—Production
- C12N2330/50—Biochemical production, i.e. in a transformed host cell
- C12N2330/51—Specially adapted vectors
-
- 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
- C12N2510/00—Genetically modified cells
- C12N2510/02—Cells for production
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
El objetivo de la invención es incrementar el rendimiento de producto de proteína en un proceso de fermentación microbiano. Dicho objetivo se logra mediante un método en el cual se introduce una construcción de expresión en un microorganismo de la especie Bacillus pumilus, cuya construcción de expresión comprende un promotor y un ácido nucleico que codifica para la proteína y la proteína se expresa en tal construcción de expresión.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE102012201297A DE102012201297A1 (de) | 2012-01-31 | 2012-01-31 | Expressionsverfahren |
| PCT/EP2013/051656 WO2013113689A1 (de) | 2012-01-31 | 2013-01-29 | Expressionsverfahren |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| MX2014009118A true MX2014009118A (es) | 2015-03-03 |
| MX351354B MX351354B (es) | 2017-10-11 |
Family
ID=47683703
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| MX2014009118A MX351354B (es) | 2012-01-31 | 2013-01-29 | Método de expresión. |
Country Status (15)
| Country | Link |
|---|---|
| US (1) | US9725704B2 (es) |
| EP (1) | EP2809816B9 (es) |
| JP (1) | JP6335793B2 (es) |
| KR (1) | KR20150000876A (es) |
| CN (1) | CN104471066B (es) |
| BR (1) | BR112014018534A8 (es) |
| CA (1) | CA2862731A1 (es) |
| DE (1) | DE102012201297A1 (es) |
| DK (1) | DK2809816T3 (es) |
| ES (1) | ES2703753T3 (es) |
| IN (1) | IN2014DN07148A (es) |
| MX (1) | MX351354B (es) |
| PL (1) | PL2809816T3 (es) |
| RU (1) | RU2644199C2 (es) |
| WO (1) | WO2013113689A1 (es) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE102011118032A1 (de) | 2011-05-31 | 2012-12-06 | Henkel Ag & Co. Kgaa | Expressionsvektoren zur verbesserten Proteinsekretion |
| DE102012215107A1 (de) | 2012-08-24 | 2014-02-27 | Basf Se | Festes Geschirrspülmittel mit verbesserter Proteaseleistung |
| CN105483100B (zh) * | 2016-01-13 | 2019-08-13 | 福建农林大学 | 一种诱导海洋微生物发酵产κ-卡拉胶酶的组合诱导物 |
| WO2020113365A1 (zh) * | 2018-12-03 | 2020-06-11 | 清华大学 | 一种高产脂肽的基因工程菌及其应用 |
| EP3938504A1 (en) * | 2019-03-11 | 2022-01-19 | Basf Se | Amylases and methods for making and using them |
| MX2021011358A (es) | 2019-03-19 | 2021-10-13 | Bayer Cropscience Lp | Proteinas de fusion, bacterias recombinantes, y fragmentos de exosporio para el control de plagas y salud vegetal. |
| CN111378597B (zh) * | 2020-01-17 | 2022-07-12 | 合肥工业大学 | 一株可用于脱锰的锰氧化细菌及其应用 |
| CN111378596B (zh) * | 2020-01-17 | 2022-07-12 | 合肥工业大学 | 一株耐酸且兼性厌氧的锰氧化细菌及其应用 |
Family Cites Families (38)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0787790B2 (ja) | 1987-03-04 | 1995-09-27 | 三菱瓦斯化学株式会社 | 新規な遺伝子dnaおよびアルカリ性プロテア−ゼの製造法 |
| DK0493398T3 (da) | 1989-08-25 | 2000-05-22 | Henkel Research Corp | Alkalisk, proteolytisk enzym og fremgangsmåde til fremstilling deraf |
| EP0501765A1 (en) | 1991-03-01 | 1992-09-02 | Takeda Chemical Industries, Ltd. | Method of producing D-ribose |
| DE69133633D1 (de) | 1991-06-11 | 2010-07-08 | Genencor Int | Cellulasezusammensetzungen mit einem Defizit an Komponenten des Typs-CBH I enthaltende Reinigungsmittelzusammensetzungen |
| US20030054534A1 (en) * | 1992-07-02 | 2003-03-20 | Novozymes A/S | DNA constructs and methods of producing cellulytic enzymes |
| GB2279955B (en) * | 1993-07-15 | 1998-02-18 | Solvay | Xylanase derived from a Bacillus species, expression vectors for such xylanase and other proteins, host organisms therefor and use thereof |
| EP0701605B2 (en) | 1994-02-24 | 2017-02-22 | Basf Se | Improved enzymes and detergents containing them |
| US5646044A (en) | 1995-03-02 | 1997-07-08 | Henkel Kommanditgesellschaft Auf Aktien | Expression systems for the production of target proteins in bacillus |
| CN101955921A (zh) | 1995-03-17 | 2011-01-26 | 诺沃奇梅兹有限公司 | 新的内切葡聚糖酶 |
| ES2525677T3 (es) | 1995-10-17 | 2014-12-29 | Ab Enzymes Oy | Celulasas, genes que las codifican y usos de las mismas |
| DE69735767T2 (de) | 1996-09-17 | 2007-04-05 | Novozymes A/S | Cellulasevarianten |
| US5958728A (en) * | 1996-11-18 | 1999-09-28 | Novo Nordiskbiotech, Inc. | Methods for producing polypeptides in mutants of bacillus cells |
| JP4068155B2 (ja) * | 1996-11-18 | 2008-03-26 | ノボザイムス,インコーポレイティド | バシラス細胞のサーファクチン突然変異株におけるポリペプチドの製造方法 |
| DE19713852A1 (de) | 1997-04-04 | 1998-10-08 | Henkel Kgaa | Aktivatoren für Persauerstoffverbindungen in Wasch- und Reinigungsmitteln |
| US5955310A (en) * | 1998-02-26 | 1999-09-21 | Novo Nordisk Biotech, Inc. | Methods for producing a polypeptide in a bacillus cell |
| WO2000053778A1 (fr) | 1999-03-08 | 2000-09-14 | Showa Sangyo Co., Ltd. | Promoteurs |
| AU781258B2 (en) | 1999-03-31 | 2005-05-12 | Novozymes A/S | Polypeptides having alkaline alpha-amylase activity and nucleic acids encoding same |
| JP2003518258A (ja) | 1999-12-23 | 2003-06-03 | ファルマシア・アンド・アップジョン・カンパニー | アミロイドβまたはその凝集体の標的としてのナトリウムチャネルの使用に基づくアッセイ方法ならびに診断および治療方法 |
| DK1305432T3 (da) | 2000-08-04 | 2010-12-06 | Genencor Int | Muterede trichoderma reesei EGIII-cellulaser, DNA, der koder for sådanne EGIII-sammensætninger og fremgangsmåder til at frembringe samme |
| DE10121463A1 (de) * | 2001-05-02 | 2003-02-27 | Henkel Kgaa | Neue Alkalische Protease-Varianten und Wasch- und Reinigungsmittel enthaltend diese neuen Alkalischen Protease-Varianten |
| JP4242761B2 (ja) | 2001-06-06 | 2009-03-25 | ノボザイムス アクティーゼルスカブ | エンド−β−1,4−グルカナーゼ |
| US20040072354A1 (en) * | 2001-06-29 | 2004-04-15 | Bedzyk Laura A. | Natural promoters for gene expression and metabolic monitoring in bacillus species |
| DE10131441A1 (de) | 2001-06-29 | 2003-01-30 | Henkel Kgaa | Eine neue Gruppe von alpha-Amylasen sowie ein Verfahren zur Identifizierung und Gewinnung neuer alpha-Amylasen |
| DE10163748A1 (de) | 2001-12-21 | 2003-07-17 | Henkel Kgaa | Neue Glykosylhydrolasen |
| KR100879908B1 (ko) * | 2001-12-21 | 2009-01-21 | 노보자임스 바이오폴리머 에이/에스 | 재조합체 숙주 세포에서 히알루로난의 제조 방법 |
| DE10260903A1 (de) | 2002-12-20 | 2004-07-08 | Henkel Kgaa | Neue Perhydrolasen |
| CN1981035B (zh) | 2003-12-03 | 2011-06-08 | 金克克国际有限公司 | 过水解酶 |
| EP2287178A3 (en) * | 2004-01-09 | 2011-05-18 | Novozymes, Inc. | Bacillus licheniformis YvmC inactivation |
| DE102004029475A1 (de) | 2004-06-18 | 2006-01-26 | Henkel Kgaa | Neues enzymatisches Bleichsystem |
| CN100372937C (zh) | 2005-07-21 | 2008-03-05 | 复旦大学 | 短小芽孢杆菌表达系统 |
| DE102006038448A1 (de) | 2005-12-28 | 2008-02-21 | Henkel Kgaa | Enzym-haltiges Reinigungsmittel |
| DE102006022224A1 (de) | 2006-05-11 | 2007-11-15 | Henkel Kgaa | Subtilisin aus Bacillus pumilus und Wasch- und Reinigungsmittel enthaltend dieses neue Subtilisin |
| DE102006032507A1 (de) * | 2006-07-12 | 2008-01-17 | Henkel Kgaa | UV-sensitive Sicherheitsstämme für die biotechnolgische Produktion |
| DE102007003143A1 (de) | 2007-01-16 | 2008-07-17 | Henkel Kgaa | Neue Alkalische Protease aus Bacillus gibsonii und Wasch- und Reinigungsmittel enthaltend diese neue Alkalische Protease |
| KR101109985B1 (ko) | 2007-08-10 | 2012-09-07 | 요시치카 카메야마 | 위액촉진 작용을 갖는 신규 미생물 및 그 분비 조성물 |
| JP5629584B2 (ja) * | 2007-12-21 | 2014-11-19 | ダニスコ・ユーエス・インク | バチルス内での促進されたタンパク質生成 |
| JPWO2009087826A1 (ja) * | 2008-01-04 | 2011-05-26 | 天野エンザイム株式会社 | シュウ酸脱炭酸酵素の生産に利用される組換え発現プラスミドベクター及び組換え菌、並びに組換えシュウ酸脱炭酸酵素の生産方法 |
| DE102008052529A1 (de) * | 2008-10-21 | 2010-04-22 | Henkel Ag & Co. Kgaa | Expressionsverstärkte Nukleinsäuren |
-
2012
- 2012-01-31 DE DE102012201297A patent/DE102012201297A1/de not_active Withdrawn
-
2013
- 2013-01-29 DK DK13703546.5T patent/DK2809816T3/en active
- 2013-01-29 WO PCT/EP2013/051656 patent/WO2013113689A1/de not_active Ceased
- 2013-01-29 EP EP13703546.5A patent/EP2809816B9/de not_active Not-in-force
- 2013-01-29 JP JP2014553753A patent/JP6335793B2/ja not_active Expired - Fee Related
- 2013-01-29 CN CN201380007050.6A patent/CN104471066B/zh not_active Expired - Fee Related
- 2013-01-29 BR BR112014018534A patent/BR112014018534A8/pt not_active IP Right Cessation
- 2013-01-29 ES ES13703546T patent/ES2703753T3/es active Active
- 2013-01-29 US US14/374,556 patent/US9725704B2/en not_active Expired - Fee Related
- 2013-01-29 KR KR1020147023866A patent/KR20150000876A/ko not_active Abandoned
- 2013-01-29 MX MX2014009118A patent/MX351354B/es active IP Right Grant
- 2013-01-29 IN IN7148DEN2014 patent/IN2014DN07148A/en unknown
- 2013-01-29 PL PL13703546T patent/PL2809816T3/pl unknown
- 2013-01-29 RU RU2014134728A patent/RU2644199C2/ru not_active IP Right Cessation
- 2013-01-29 CA CA2862731A patent/CA2862731A1/en not_active Abandoned
Also Published As
| Publication number | Publication date |
|---|---|
| BR112014018534A2 (pt) | 2018-09-04 |
| ES2703753T3 (es) | 2019-03-12 |
| US20140370569A1 (en) | 2014-12-18 |
| RU2644199C2 (ru) | 2018-02-08 |
| WO2013113689A1 (de) | 2013-08-08 |
| EP2809816B1 (de) | 2018-10-03 |
| BR112014018534A8 (pt) | 2019-01-29 |
| JP6335793B2 (ja) | 2018-05-30 |
| US9725704B2 (en) | 2017-08-08 |
| DK2809816T3 (en) | 2019-01-28 |
| RU2014134728A (ru) | 2016-03-27 |
| DE102012201297A1 (de) | 2013-08-01 |
| MX351354B (es) | 2017-10-11 |
| CN104471066B (zh) | 2019-09-03 |
| PL2809816T3 (pl) | 2019-04-30 |
| CN104471066A (zh) | 2015-03-25 |
| JP2015505463A (ja) | 2015-02-23 |
| EP2809816B9 (de) | 2019-02-27 |
| KR20150000876A (ko) | 2015-01-05 |
| CA2862731A1 (en) | 2013-08-08 |
| EP2809816A1 (de) | 2014-12-10 |
| IN2014DN07148A (es) | 2015-04-24 |
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