DK2465341T3 - EPSPS-mutanter - Google Patents
EPSPS-mutanter Download PDFInfo
- Publication number
- DK2465341T3 DK2465341T3 DK12152493.8T DK12152493T DK2465341T3 DK 2465341 T3 DK2465341 T3 DK 2465341T3 DK 12152493 T DK12152493 T DK 12152493T DK 2465341 T3 DK2465341 T3 DK 2465341T3
- Authority
- DK
- Denmark
- Prior art keywords
- epsps
- leu
- plant
- ala
- thr
- Prior art date
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Classifications
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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
- 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
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
- C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
- C12N15/8274—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for herbicide resistance
- C12N15/8275—Glyphosate
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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
- 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)
- C12N15/8201—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation
- C12N15/8213—Targeted insertion of genes into the plant genome by homologous recombination
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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
- C12N9/10—Transferases (2.)
- C12N9/1085—Transferases (2.) transferring alkyl or aryl groups other than methyl groups (2.5)
- C12N9/1092—3-Phosphoshikimate 1-carboxyvinyltransferase (2.5.1.19), i.e. 5-enolpyruvylshikimate-3-phosphate synthase
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- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- General Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Cell Biology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Medicinal Chemistry (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Enzymes And Modification Thereof (AREA)
- Peptides Or Proteins (AREA)
Claims (14)
1. Fremgangsmåde til fremstilling afen ikke-transgen, herbicidresistent eller -tolerant plante, hvilken fremgangsmåde omfatter: indføring af en rekombinagen oligonukleobase med en targeted mutation i EPSPS-genet i planteceller til fremstilling af planteceller med et EPSPS-mutantgen, som eksprimerer et EPSPS-protein, som er muteret ved amino-syreposition Thri79 og Proi83 i et Arabidopsis-EPSPS-protein (AF360224) eller ved en analog aminosyrerest i et EPSPS-protein af en anden art, hvor Thri/g er ændret til Ile, og Proi83 er ændret til Thr; udvælgelse af en plantecelle, der udviser en forbedret tolerance i forhold til glyphosat sammenlignet med en tilsvarende plantecelle af vildtypen; og regenerering af en ikke-transgen herbicidresistent eller -tolerant plante med et muteret EPSPS-gen fra den udvalgte plantecelle.
2. Fremgangsmåde til fremstilling afen ikke-transgen, herbicidresistent eller -tolerant plante, hvilken fremgangsmåde omfatter: indføring af en rekombinagen oligonukleobase med en targeted mutation i EPSPS-genet i planteceller til fremstilling af planteceller med et EPSPS-mutantgen, som eksprimerer et EPSPS-protein, som er muteret ved amino-syreposition Thri79 og Proi83 i et Arabidopsis-EPSPS-protein (AF360224) eller ved en analog aminosyrerest i et EPSPS-protein af en anden art, hvor Thri79 er ændret til Ile, og Proi83 er ændret til Thr; identifikation af en plantecelle med et EPSPS-mutantprotein, som udviser i det væsentlige samme katalytiske aktivitet som et EPSPS-protein af vildtypen, og som udviser den aktivitet selv i nærvær af glyphosat; og regenerering af en ikke-transgen herbicidresistent eller -tolerant plante med et muteret EPSPS-gen fra plantecellen.
3. Fremgangsmåde ifølge krav 1 eller 2, hvorved den rekombinagene oligonukleobase indføres ved elektroporese.
4. Fremgangsmåde ifølge et af kravene 1 til 3, hvorved plantecellerne udvælges fra gruppen bestående af majs, hvede, ris, byg, sojabønne, bomuld, sukkerroe, raps, canola, hør, solsikke, kartoffel, tobak, tomat, lucerne, poppel, fyr, eukalyptus, æble, salat, ærter, linser, drue, plænegræs (turf grasses) og Brassica sp.
5. Fremgangsmåde ifølge et af kravene 1 til 3, hvorved aminosyrepositioner-ne er Thrio2 og Ργοιοθ i EPSPS-proteinet af Zea mays; Thri/4 og Proue i EPSPS-proteinet af et Brassica sp; Thri74 og Proi7s i EPSPS-proteinet af Petunia hybrida; eller Thrus og Proi82 i EPSPS-proteinet af Arabidopsis (NM 130093).
6. Fremgangsmåde til fremstilling af en ikke-transgen E. coli-celle med et EPSPS-mutantgen, hvilken fremgangsmåde omfatter: indføring af en rekombinagen oligonukleobase med en targeted mutation i EPSPS-genet i E. coli-celler til fremstilling af E. coli-celler med et EPSPS-mutantgen, som eksprimerer et EPSPS-protein, som er muteret ved amino-syreposition Thr97 og Proioi, hvor Thr97 er ændret til Ile, og Proioi er ændret til Thr; identifikation afen E. co//-cellekoloni med en i det væsentlige normal vækst i nærvær af glyphosat; og isolering af en eller flere E. coli-celler, som indeholder EPSPS-mutantgenet.
7. Fremgangsmåde ifølge et af de foregående krav, hvorved den rekombi-nagene oligonukleobase er et blandet duplex-nukleotid eller en SSOMV.
8. Fremgangsmåde ifølge krav 7, hvorved det blandede duplex-nukleotid indeholder en første homolog region, som har en sekvens, som er identisk med sekvensen på mindst 6 basepar af det første fragment af EPSPS-targetgenet, og en anden homolog region, som har en sekvens, som er identisk med sekvensen på mindst 6 basepar af et andet fragment af EPSPS-targetgenet, og en mellemliggende region, som indeholder mindst en nu-kleobase, som er heterolog i forhold til EPSPS-targetgenet, hvor den mellemliggende region forbinder den første og anden homologe region.
9. Herbicidresistent plante, som eksprimerer et EPSPS-mutantgenprodukt, hvor EPSPS-genet er muteret ved positioner til ændring ved aminosyreposi-tion Thrug og Proi83 i et Arabidopsis EPSPS-protein (AF360224) eller ved en analog aminosyrerest i et EPSPS-homolog, hvor Thr^g er ændret til Ile, og Proi83 er ændret til Ala.
10. Plante ifølge krav 9, hvor planten er udvalgt fra gruppen bestående af majs, hvede, ris, byg, sojabønne, bomuld, sukkerroe, raps, canola, hør, solsikke, kartoffel, tobak, tomat, lucerne, poppel, fyr, eukalyptus, æble, salat, ærter, linser, drue, plænegræs (turf grasses) og Brassica sp.
11. Plante ifølge krav 9 til 10, hvor aminosyrepositionerne er Thr102 og Ργοιοθ i EPSPS-proteinet af Zea mays; Thri/4 og Prous i EPSPS-proteinet af et Brassica sp; Thri/4 og Prous i EPSPS-proteinet af Petunia hybrida; eller Thri/s og Proi82 i EPSPS-proteinet af Arabidopsis (NM 130093).
12. EPSPS-mutantprotein omfattende aminosyresekvensen af E. coli-EPSPS-genet vist i FIG. 1 eller af et EPSPS-homolog, hvor aminosyrepositi-on Thr97 og Proioi er ændret, hvor Thr97 er ændret til Ile, og Proioi er ændret til Thr, hvor EPSPS-mutantproteinet har en øget modstand eller tolerance i forhold til et phosphonomethylglycinherbicid. 13. E. coli-mutantcelle, som eksprimerer et EPSPS-mutantgenprodukt, hvor EPSPS-genet er muteret til at ændre aminosyreposition Thr97 og Proioi i genproduktet, hvor Thr97 er ændret til Ile, og Proioi er ændret til Thr.
14. Fremgangsmåde ifølge et af kravene 1 til 3, hvorved den herbicidresistente eller -tolerante plante udvælges fra gruppen bestående af majs, hvede, sukkerroe, kartoffel, raps og canola; og hvor herbicidet erglyphosat.
15. Plante ifølge krav 9, hvor den herbicidresistente eller -tolerante plante er udvalgt fra gruppen bestående af majs, hvede, sukkerroe, kartoffel, raps og canola; og hvor herbicidet er glyphosat.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US75843906P | 2006-01-12 | 2006-01-12 | |
| EP07716464A EP1978799A4 (en) | 2006-01-12 | 2007-01-10 | MUTANT EPSPS |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| DK2465341T3 true DK2465341T3 (da) | 2015-03-02 |
Family
ID=38288103
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DK12152491.2T DK2465340T3 (da) | 2006-01-12 | 2007-01-10 | EPSPS-mutanter |
| DK12152493.8T DK2465341T3 (da) | 2006-01-12 | 2007-01-10 | EPSPS-mutanter |
Family Applications Before (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DK12152491.2T DK2465340T3 (da) | 2006-01-12 | 2007-01-10 | EPSPS-mutanter |
Country Status (14)
| Country | Link |
|---|---|
| US (3) | US8268622B2 (da) |
| EP (4) | EP2465341B1 (da) |
| JP (1) | JP2009523418A (da) |
| AU (1) | AU2007207813B2 (da) |
| CA (3) | CA2923605C (da) |
| DK (2) | DK2465340T3 (da) |
| ES (2) | ES2532112T3 (da) |
| NZ (4) | NZ570009A (da) |
| PL (2) | PL2465341T3 (da) |
| PT (2) | PT2465340E (da) |
| RU (1) | RU2441366C2 (da) |
| SI (2) | SI2465341T1 (da) |
| UA (1) | UA99594C2 (da) |
| WO (1) | WO2007084294A2 (da) |
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