WO2012142371A1 - Synthetic genes - Google Patents
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- WO2012142371A1 WO2012142371A1 PCT/US2012/033458 US2012033458W WO2012142371A1 WO 2012142371 A1 WO2012142371 A1 WO 2012142371A1 US 2012033458 W US2012033458 W US 2012033458W WO 2012142371 A1 WO2012142371 A1 WO 2012142371A1
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- sequence
- dna sequence
- synthetic dna
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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
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
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- 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
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0065—Oxidoreductases (1.) acting on hydrogen peroxide as acceptor (1.11)
-
- 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
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0069—Oxidoreductases (1.) acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)
-
- 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
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
- C12N9/0083—Miscellaneous (1.14.99)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y111/00—Oxidoreductases acting on a peroxide as acceptor (1.11)
- C12Y111/01—Peroxidases (1.11.1)
- C12Y111/01015—Peroxiredoxin (1.11.1.15)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y113/00—Oxidoreductases acting on single donors with incorporation of molecular oxygen (oxygenases) (1.13)
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- 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
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/10—Cells modified by introduction of foreign genetic material
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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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/146—Genetically Modified [GMO] plants, e.g. transgenic plants
Definitions
- Also provided is a method for herbicide tolerance in a plant which comprises expressing a synthetic DNA sequence of the invention in the plant where the synthetic DNA sequence encodes a known herbicide tolerance enzyme, for example the aryloxyalkanoate dioxygenase (AAD1) see WO/2005/107437, or phosphinothricin acetytransferase, or 5- enolpymvylshikimate-3 -phosphate synthase enzymes.
- AAD1 aryloxyalkanoate dioxygenase
- AAD1 aryloxyalkanoate dioxygenase
- phosphinothricin acetytransferase or 5- enolpymvylshikimate-3 -phosphate synthase enzymes.
- SEQ ID NO: 14 is Bacillus thuringiensis Cry35Abl toxin sequence.
- SEQ ID NO:37 is the native DNA sequence encoding Sphingobiurn herbicidovorans AAD1 protein.
- SEQ ID NO:46 is Aspergillus nidulans delta-9 fatty acid desaturase protein sequence.
- SEQ ID NO:47 is a synthetic DNA sequence in accordance with the invention encoding Aspergillus nidulans delta-9 fatty acid desaturase protein using codons optimized for maize and with sequences identified in Table 2 removed and Table 1 sequences are maintained.
- T-DNA-containing vectors for the transformation of plant cells has been intensively researched and sufficiently described in European Patent No. EP120516B1; Lee and Gelvin (2008) Plant Physiol. 146:325-332; Fraley et al. (1986) Crit. Rev. Plant Sci. 4: 1-46; and An et al. (1985) EMBO J. 4:277-284; and is well established in the field. Additionally, fusion of plant protoplasts with liposomes containing the DNA to be delivered, direct injection of the DNA, biolistics transformation (microparticle bombardment), or electroporation, as well as other possible methods, may be employed.
- DNA of SEQ ID NO:5 is synthesized, and expression levels observed in plant cells transformed to express this sequence are compared with expression levels observed in plant cells transformed to express SEQ ID NO: l and SEQ ID NO:3.
- a construct for use in expressing the synthetic coding region of SEQ ID NO:5 is made by combining the synthetic coding region of SEQ ID NO: 5 with a 5' non-translated region comprising a promoter that functions in plant cells and a 3' non-translated region comprising a transcription termination and polyadenylation sequence.
- Table 7 Table 1 sequences found in the native Cry34Abl coding region (SEQ ID NO:7) and the redesi ned version SE ID NO: 11
- the presence of the synthetic gene insert in the binary plant transformation vector is performed by PCR analysis using vector-specific primers with template plasmid DNA prepared from selected Agrobacterium colonies.
- the cell pellet from a 4 mL aliquot of a 15 mL overnight culture grown in YEP with spectinomycin and streptomycin as before is extracted using Qiagen Spin® Mini Preps, performed per manufacturer's instructions. Plasmid DNA from the binary vector used in the Agrobacterium electroporation transformation is included as a control.
- the PCR reaction is completed using Taq DNA polymerase from Invitrogen per manufacture's instructions at 0.5X concentrations.
- the plasmid structure of the binary plant transformation vector containing the synthetic gene insert is performed by restriction digest fingerprint mapping of plasmid DNA prepared from candidate Agrobacterium isolates by standard molecular biology methods well known to those skilled in the art of Agrobacterium manipulation.
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- General Engineering & Computer Science (AREA)
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- Biotechnology (AREA)
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- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Nutrition Science (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Mycology (AREA)
- Crystallography & Structural Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Toxicology (AREA)
- Pest Control & Pesticides (AREA)
- Agronomy & Crop Science (AREA)
- Insects & Arthropods (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
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Abstract
Description
Claims
Priority Applications (14)
Application Number | Priority Date | Filing Date | Title |
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EP12771427.7A EP2697363B1 (en) | 2011-04-15 | 2012-04-13 | Synthetic genes |
NZ616646A NZ616646A (en) | 2011-04-15 | 2012-04-13 | Synthetic genes |
CN201280028762.1A CN103597070B (en) | 2011-04-15 | 2012-04-13 | Synthetic gene |
KR1020137029911A KR20140026479A (en) | 2011-04-15 | 2012-04-13 | Synthetic genes |
JP2014505322A JP2014515609A (en) | 2011-04-15 | 2012-04-13 | Synthetic gene |
CA2832834A CA2832834C (en) | 2011-04-15 | 2012-04-13 | Synthetic dna molecules for expressing a protein of interest |
AU2012242735A AU2012242735B2 (en) | 2011-04-15 | 2012-04-13 | Synthetic genes |
MX2013012052A MX348009B (en) | 2011-04-15 | 2012-04-13 | Synthetic genes. |
BR112013026179-0A BR112013026179A2 (en) | 2011-04-15 | 2012-04-13 | synthetic genes |
UAA201313262A UA115526C2 (en) | 2011-04-15 | 2012-04-13 | Synthetic genes |
RU2013150784A RU2632571C2 (en) | 2011-04-15 | 2012-04-13 | Synthetic genes |
EP18183438.3A EP3406708B1 (en) | 2011-04-15 | 2012-04-13 | Synthetic genes |
IL228849A IL228849A0 (en) | 2011-04-15 | 2013-10-13 | Synthetic genes |
ZA2013/07806A ZA201307806B (en) | 2011-04-15 | 2013-10-18 | Synthetic genes |
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US201161475921P | 2011-04-15 | 2011-04-15 | |
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US (4) | US9427003B2 (en) |
EP (2) | EP2697363B1 (en) |
JP (1) | JP2014515609A (en) |
KR (1) | KR20140026479A (en) |
CN (1) | CN103597070B (en) |
AR (1) | AR086006A1 (en) |
AU (1) | AU2012242735B2 (en) |
BR (1) | BR112013026179A2 (en) |
CA (2) | CA3055700C (en) |
CL (1) | CL2013002992A1 (en) |
CO (1) | CO6801652A2 (en) |
IL (1) | IL228849A0 (en) |
MX (1) | MX348009B (en) |
MY (1) | MY161583A (en) |
NZ (1) | NZ616646A (en) |
RU (1) | RU2632571C2 (en) |
UA (1) | UA115526C2 (en) |
UY (1) | UY34014A (en) |
WO (1) | WO2012142371A1 (en) |
ZA (1) | ZA201307806B (en) |
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2012
- 2012-04-11 UY UY0001034014A patent/UY34014A/en not_active Application Discontinuation
- 2012-04-12 AR ARP120101262A patent/AR086006A1/en unknown
- 2012-04-13 KR KR1020137029911A patent/KR20140026479A/en not_active Application Discontinuation
- 2012-04-13 UA UAA201313262A patent/UA115526C2/en unknown
- 2012-04-13 JP JP2014505322A patent/JP2014515609A/en active Pending
- 2012-04-13 MX MX2013012052A patent/MX348009B/en active IP Right Grant
- 2012-04-13 MY MYPI2013701935A patent/MY161583A/en unknown
- 2012-04-13 CA CA3055700A patent/CA3055700C/en active Active
- 2012-04-13 CA CA2832834A patent/CA2832834C/en active Active
- 2012-04-13 EP EP12771427.7A patent/EP2697363B1/en active Active
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- 2012-04-13 NZ NZ616646A patent/NZ616646A/en not_active IP Right Cessation
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- 2012-04-13 RU RU2013150784A patent/RU2632571C2/en not_active IP Right Cessation
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- 2012-04-13 BR BR112013026179-0A patent/BR112013026179A2/en not_active Application Discontinuation
- 2012-04-13 EP EP18183438.3A patent/EP3406708B1/en active Active
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2013
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