ES2653727T3 - Desaturasas y proceso para la producción de ácidos grasos poliinsaturados en organismos transgénicos - Google Patents
Desaturasas y proceso para la producción de ácidos grasos poliinsaturados en organismos transgénicos Download PDFInfo
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- ES2653727T3 ES2653727T3 ES13180237.3T ES13180237T ES2653727T3 ES 2653727 T3 ES2653727 T3 ES 2653727T3 ES 13180237 T ES13180237 T ES 13180237T ES 2653727 T3 ES2653727 T3 ES 2653727T3
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- 230000009261 transgenic effect Effects 0.000 title description 2
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 title 1
- 230000000694 effects Effects 0.000 abstract description 6
- 108090000623 proteins and genes Proteins 0.000 abstract description 5
- 108010033653 omega-3 fatty acid desaturase Proteins 0.000 abstract description 4
- 150000007523 nucleic acids Chemical group 0.000 abstract 6
- 108091028043 Nucleic acid sequence Proteins 0.000 abstract 5
- 229920001184 polypeptide Polymers 0.000 abstract 4
- 102000004196 processed proteins & peptides Human genes 0.000 abstract 4
- 108090000765 processed proteins & peptides Proteins 0.000 abstract 4
- 125000003275 alpha amino acid group Chemical group 0.000 abstract 2
- 241001085796 Hyaloperonospora Species 0.000 abstract 1
- 230000003321 amplification Effects 0.000 abstract 1
- 238000003199 nucleic acid amplification method Methods 0.000 abstract 1
- 108020004707 nucleic acids Proteins 0.000 abstract 1
- 102000039446 nucleic acids Human genes 0.000 abstract 1
- 230000003071 parasitic effect Effects 0.000 abstract 1
- 108091033319 polynucleotide Proteins 0.000 abstract 1
- 102000040430 polynucleotide Human genes 0.000 abstract 1
- 239000002157 polynucleotide Substances 0.000 abstract 1
- 102000004169 proteins and genes Human genes 0.000 abstract 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 7
- 229930195729 fatty acid Natural products 0.000 description 7
- 239000000194 fatty acid Substances 0.000 description 7
- 150000004665 fatty acids Chemical class 0.000 description 5
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 2
- 235000020673 eicosapentaenoic acid Nutrition 0.000 description 2
- 229960005135 eicosapentaenoic acid Drugs 0.000 description 2
- JAZBEHYOTPTENJ-UHFFFAOYSA-N eicosapentaenoic acid Natural products CCC=CCC=CCC=CCC=CCC=CCCCC(O)=O JAZBEHYOTPTENJ-UHFFFAOYSA-N 0.000 description 2
- -1 eicosapentaenoic acid fatty acid Chemical class 0.000 description 2
- 235000020978 long-chain polyunsaturated fatty acids Nutrition 0.000 description 2
- 101150053747 D12Des gene Proteins 0.000 description 1
- 101100330364 Rebecca salina D5Des gene Proteins 0.000 description 1
- JAZBEHYOTPTENJ-JLNKQSITSA-N all-cis-5,8,11,14,17-icosapentaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O JAZBEHYOTPTENJ-JLNKQSITSA-N 0.000 description 1
- 235000020669 docosahexaenoic acid Nutrition 0.000 description 1
- MBMBGCFOFBJSGT-KUBAVDMBSA-N docosahexaenoic acid Natural products CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCC(O)=O MBMBGCFOFBJSGT-KUBAVDMBSA-N 0.000 description 1
- 229940090949 docosahexaenoic acid Drugs 0.000 description 1
- 230000037353 metabolic pathway Effects 0.000 description 1
- 235000020660 omega-3 fatty acid Nutrition 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
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- 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
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6409—Fatty acids
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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
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
- C12N15/8243—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
- C12N15/8247—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine involving modified lipid metabolism, e.g. seed oil composition
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
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- 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
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
- C12N9/0083—Miscellaneous (1.14.99)
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- 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
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6409—Fatty acids
- C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
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- 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
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6409—Fatty acids
- C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
- C12P7/6431—Linoleic acids [18:2[n-6]]
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- 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
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6409—Fatty acids
- C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
- C12P7/6432—Eicosapentaenoic acids [EPA]
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6409—Fatty acids
- C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
- C12P7/6434—Docosahexenoic acids [DHA]
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- 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
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6436—Fatty acid esters
- C12P7/6445—Glycerides
- C12P7/6472—Glycerides containing polyunsaturated fatty acid [PUFA] residues, i.e. having two or more double bonds in their backbone
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- 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
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6436—Fatty acid esters
- C12P7/6445—Glycerides
- C12P7/6481—Phosphoglycerides
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y114/00—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14)
- C12Y114/19—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14) with oxidation of a pair of donors resulting in the reduction of molecular oxygen to two molecules of water (1.14.19)
- C12Y114/19001—Stearoyl-CoA 9-desaturase (1.14.19.1), i.e. DELTA9-desaturase
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Nutrition Science (AREA)
- Cell Biology (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Enzymes And Modification Thereof (AREA)
- Cosmetics (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
Un polinucleótido que comprende una secuencia de ácidos nucleicos seleccionada del grupo que consiste de: (a) una secuencia de ácidos nucleicos como se muestra en la SEQ ID NO. 50 o 51; (b) una secuencia de ácidos nucleicos que codifica un polipéptido que caracteriza una secuencia de aminoácidos como se muestra en cualquiera de la SEQ ID NO. 52; (c) una secuencia de ácidos nucleicos que codifica un polipéptido con una secuencia de aminoácidos con por lo menos 75 % de identidad con la SEQ ID NO. 52, y que codifica un polipéptido con una actividad de omega-3- desaturasa; y (d) una secuencia de ácidos nucleicos obtenible u obtenida mediante la amplificación de ácido nucleico del material genético de Hyaloperonospora parasitica usando el primer par - SEQ ID NO. 53 y SEQ ID NO. 55, o - SEQ ID NO. 54 y SEQ ID NO. 56 y que codifica un polipéptido con una actividad de omega-3-desaturasa.
Description
Tabla 4A: Alimentación de levaduras
- Nombre de la muestra/ carga de ácido graso
- Etapa de reacción observada Índice de conversión (%) Actividad observada Fig.
- Sustrato (% de contenido de ácido graso total)
- Producto (% de contenido de ácido graso total) Esperado observado
- Vector vacío pYES2 / 18: 2n-9
- 18:2n-6 25,3 18:3n-3 0,0 - - - 4a
- d15Des (Ch)/18: 2n-9
- 18:2n-6 6,3 18:3n-3 5,7 > 0 47,5 ∆15-des. 4b
- d15Des (Cy)/18: 2n-9 ω3Des (Hp)/18: 2n-9 ω3Des (Hp)/20: 3n-6 ω3Des (Hp)/20: 4n-6
- 18:2n-6 18:2n-6 20:3n-6 20:4n-6 13,0 12,6 6,3 5,7 18:3n-3 18:3n-3 20:4n-3 20:5n-3 0,9 0,0 0,6 1,8 > 0 > 0 > 0 > 0 6,4 0,0 8,6 24,0 ∆15-des. -ω3-Des ω3-Des 4c
A modo de control para el ensayo para la actividad ∆ 15-desaturasa, las levaduras se transforman con el vector vacío pYES, se carga el ácido graso 18:2 n-6 y el perfil de ácidos grasos se analiza (Figura 2A). En comparación, el 5 ácido graso adicional 18:3 n-3 se puede observar en levaduras que expresan las desaturasas d15Des (Ch) y d15Des(Cy) (Figura 2B, 2C). Estas desaturasas que se han ensayado, por lo tanto, tienen actividad ∆15-desaturasa.
Ejemplo 5: Producción de Plantas transgénicas para la producción de ácidos grasos poliinsaturados de cadena larga
Para producir ácidos grasos poliinsaturados de cadena larga en plantas, diversos genes de la ruta metabólica se combinan en un vector binario. Para producir el ácido graso ácido eicosapentaenoico (20:5∆5,8,11,14,17), se 10 combinan los genes como se describe en la Tabla 5. Análogamente, Los genes como se describe en la Tabla 6 se combinan para producir el ácido graso ácido docosahexaenoico (22:06∆4,7,10,13,16,19).
Tabla 5: Combinación de genes para la producción de ácido eicosapentaenoico
- Gen
- Actividad SEQ ID NO.
- D6Des(Pir)
- ∆6-desaturasa 22
- D6Elo(Pp)
- ∆6-elongasa 31
- D5Des(Tc)
- ∆5-desaturasa 25
- ω3-Des(Pi)
- ω3-desaturasa 28
- D15Des(Ch)
- ∆12-desaturasa 2
- D15Des(Cy)
- ∆15-desaturasa 9
- D12Des(Mac)
- ∆12-/∆15-desaturasa 16
- ω 3Des(Hp)
- ω3-desaturasa 51
35
Claims (1)
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imagen1 imagen2
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP08171520 | 2008-12-12 | ||
EP08171520 | 2008-12-12 | ||
EP09151937 | 2009-02-03 | ||
EP09151937 | 2009-02-03 |
Publications (1)
Publication Number | Publication Date |
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ES2653727T3 true ES2653727T3 (es) | 2018-02-08 |
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Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
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ES13180237.3T Active ES2653727T3 (es) | 2008-12-12 | 2009-12-08 | Desaturasas y proceso para la producción de ácidos grasos poliinsaturados en organismos transgénicos |
ES09801425T Active ES2435572T3 (es) | 2008-12-12 | 2009-12-08 | Desaturasas y proceso para la producción de ácidos grasos poliinsaturados en organismos transgénicos |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
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ES09801425T Active ES2435572T3 (es) | 2008-12-12 | 2009-12-08 | Desaturasas y proceso para la producción de ácidos grasos poliinsaturados en organismos transgénicos |
Country Status (8)
Country | Link |
---|---|
US (2) | US8822662B2 (es) |
EP (2) | EP2669380B1 (es) |
AU (2) | AU2009326129B2 (es) |
CA (1) | CA2744930C (es) |
DE (1) | DE112009003708T5 (es) |
ES (2) | ES2653727T3 (es) |
PL (1) | PL2376638T3 (es) |
WO (1) | WO2010066703A2 (es) |
Families Citing this family (15)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8455035B2 (en) | 2008-04-25 | 2013-06-04 | Basf Plant Science Gmbh | Plant seed oil |
CN102317459A (zh) | 2008-11-18 | 2012-01-11 | 联邦科学技术研究组织 | 产生ω-3脂肪酸的酶和方法 |
CA2781559C (en) | 2009-11-24 | 2018-09-04 | Basf Plant Science Company Gmbh | Novel fatty acid elongase and uses thereof |
CA2780956A1 (en) | 2009-11-24 | 2011-06-03 | Basf Plant Science Company Gmbh | Novel fatty acid desaturase and uses thereof |
KR20150037771A (ko) | 2012-06-15 | 2015-04-08 | 커먼웰쓰 사이언티픽 앤 인더스트리알 리서치 오거니제이션 | 식물 세포에서 장쇄 다중불포화 지방산의 생성 |
US9873880B2 (en) | 2013-03-13 | 2018-01-23 | Dsm Nutritional Products Ag | Engineering microorganisms |
JP6726619B2 (ja) | 2013-12-18 | 2020-07-22 | コモンウェルス サイエンティフィック アンド インダストリアル リサーチ オーガナイゼーション | 長鎖多価不飽和脂肪酸を含む脂質 |
SG11201610596PA (en) | 2014-06-27 | 2017-01-27 | Commw Scient Ind Res Org | Lipid comprising docosapentaenoic acid |
CA2967132A1 (en) | 2014-11-14 | 2016-05-19 | Basf Plant Science Company Gmbh | Stabilising fatty acid compositions |
US11690825B2 (en) | 2016-03-09 | 2023-07-04 | Board Of Regents, The University Of Texas System | 20-HETE receptor (GPR75) antagonists and methods of use |
US20190240334A1 (en) | 2016-09-01 | 2019-08-08 | Instituto De Biologia Molecular E Celular - Ibmc | Cyanobacterium extracellular polymer, compositions and uses thereof |
WO2020168277A1 (en) | 2019-02-14 | 2020-08-20 | Cargill, Incorporated | Brassica plants producing elevated levels of polyunsaturated fatty acids |
EP4240144A1 (en) | 2020-11-04 | 2023-09-13 | BASF Plant Science Company GmbH | Harvest management |
CN112592831A (zh) * | 2021-01-06 | 2021-04-02 | 云南农业大学 | 一种快速分离三七圆斑病菌单个分生孢子的方法 |
AU2022246152A1 (en) | 2021-03-25 | 2023-10-05 | Basf Plant Science Company Gmbh | Fertilizer management |
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2009
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- 2009-12-08 PL PL09801425T patent/PL2376638T3/pl unknown
- 2009-12-08 ES ES09801425T patent/ES2435572T3/es active Active
- 2009-12-08 AU AU2009326129A patent/AU2009326129B2/en not_active Ceased
- 2009-12-08 WO PCT/EP2009/066569 patent/WO2010066703A2/en active Application Filing
- 2009-12-08 US US13/139,014 patent/US8822662B2/en not_active Expired - Fee Related
- 2009-12-08 CA CA2744930A patent/CA2744930C/en active Active
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US9139854B2 (en) | 2015-09-22 |
PL2376638T3 (pl) | 2014-01-31 |
AU2016202449B2 (en) | 2017-08-31 |
AU2009326129B2 (en) | 2016-05-12 |
EP2376638B1 (en) | 2013-08-14 |
US20110277043A1 (en) | 2011-11-10 |
EP2376638A2 (en) | 2011-10-19 |
CA2744930C (en) | 2018-03-20 |
WO2010066703A2 (en) | 2010-06-17 |
US20140335579A1 (en) | 2014-11-13 |
EP2669380B1 (en) | 2017-09-27 |
AU2009326129A1 (en) | 2011-07-07 |
CA2744930A1 (en) | 2010-06-17 |
US8822662B2 (en) | 2014-09-02 |
DE112009003708T5 (de) | 2012-09-13 |
EP2669380A1 (en) | 2013-12-04 |
ES2435572T3 (es) | 2013-12-20 |
WO2010066703A3 (en) | 2010-09-23 |
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