HRP20220299T1 - Ekspresijski konstrukti i postupci za genetsku modifikaciju metilotrofnog kvasca - Google Patents
Ekspresijski konstrukti i postupci za genetsku modifikaciju metilotrofnog kvasca Download PDFInfo
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- HRP20220299T1 HRP20220299T1 HRP20220299TT HRP20220299T HRP20220299T1 HR P20220299 T1 HRP20220299 T1 HR P20220299T1 HR P20220299T T HRP20220299T T HR P20220299TT HR P20220299 T HRP20220299 T HR P20220299T HR P20220299 T1 HRP20220299 T1 HR P20220299T1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/37—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi
- C07K14/39—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi from yeasts
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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
- 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
- C12N15/80—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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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/795—Porphyrin- or corrin-ring-containing peptides
- C07K14/805—Haemoglobins; Myoglobins
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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
- 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
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
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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
- C12N15/635—Externally inducible repressor mediated regulation of gene expression, e.g. tetR inducible by tetracyline
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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
- C12N15/80—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
-
- 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
- C12N2830/00—Vector systems having a special element relevant for transcription
- C12N2830/001—Vector systems having a special element relevant for transcription controllable enhancer/promoter combination
- C12N2830/002—Vector systems having a special element relevant for transcription controllable enhancer/promoter combination inducible enhancer/promoter combination, e.g. hypoxia, iron, transcription factor
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- 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
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/84—Pichia
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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
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
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- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
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- Bioinformatics & Cheminformatics (AREA)
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- General Health & Medical Sciences (AREA)
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- Molecular Biology (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Botany (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Claims (11)
1. Stanica metilotrofnog kvasca Pichia koja sadrži:
a) prvu rekombinantnu molekulu nukleinske kiseline koja sadrži egzogenu nukleinsku kiselinu koja kodira transkripcijski aktivator proteina Mxr1 koji je operativno povezan s barem jednim elementom promotorom alkohol oksidaze 1 (AOX1); i
b) drugu rekombinantnu molekulu nukleinske kiseline koja sadrži nukleinsku kiselinu koja kodira člana obitelji globina PF00042 koji je operativno povezan s barem jednim AOX1 elementom; i
c) rekombinantnu molekulu koja sadrži nukleinsku kiselinu koja kodira barem jedan polipeptid koji je uključen u biosintezu hema koji se odabire iz skupine koja se sastoji od ALA sintaze, ALA dehidrataze, porfobilinogen deaminaze, UPG III sintaze, UPG III dekarboksilaze, CPG oksidaze, PPG oksidaze i ferokelataze, a koji je operativno povezan s najmanje jednim elementom promotorom koji se aktivira metanolom.
2. Stanica kvasca prema zahtjevu 1, pri čemu je stanica metilotrofnog kvasca stanica kvasca Pichia pastoris.
3. Stanica kvasca prema bilo kojem od zahtjeva od 1 do 2, pri čemu su rekombinantne molekule nukleinske kiseline stabilno integrirane u genom stanice metilotrofnog kvasca.
4. Stanica kvasca prema bilo kojem od zahtjeva od 1 do 2, pri čemu su rekombinantne molekule nukleinske kiseline ekstrakromosomski izražene iz plazmida koji je kompetentan za replikaciju.
5. Stanica kvasca prema bilo kojem od zahtjeva od 1 do 4, pri čemu sekvencija proteina Mxr1 potječe iz kvasca Pichia pastoris.
6. Stanica kvasca prema bilo kojem od zahtjeva od 1 do 5, pri čemu transkripcijski aktivator sadrži sekvenciju koja je navedena u bazi podataka o sekvencijama GenBank, referentni broj ABD57365 ili je transkripcijski aktivator kodiran sekvencijom nukleinske kiseline koja je navedena u bazi podataka o sekvencijama GenBank, referentni broj DQ395124.
7. Stanica kvasca prema bilo kojem od zahtjeva od 1 do 6, pri čemu element promotor alkohol oksidaze 1 (AOX1) potječe iz kvasca Pichia pastoris.
8. Postupak za ekspresiju člana obitelji globina PF00042 u stanici, pri čemu se postupak sastoji od:
dobivanja stanice kvasca prema bilo kojem od zahtjeva 1 – 7; i
uzgoja stanice u uvjetima koji su prikladni za ekspresiju rekombinantnih molekula nukleinske kiseline,
što dovodi do ekspresije člana obitelji globina PF00042.
9. Postupak prema zahtjevu 8, pri čemu je član obitelji globina PF00042 biljni hemoglobin, po mogućnosti leghemoglobin, a najpoželjnije leghemoglobin koji sadrži sekvenciju koja se odabire iz skupine koja se sastoji od sekvencija SEQ ID BR:4 i SEQ ID BR:6.
10. Postupak prema bilo kojem od zahtjeva od 8 do 9, pri čemu se uvjeti u kojima se stanice uzgajaju sastoje od dodavanja željeza ili njegove farmaceutski ili metabolički prihvatljive soli.
11. Postupak prema bilo kojem od zahtjeva od 8 do 10, pri čemu se korak uzgoja sastoji od uzgajanja stanice u prisutnosti metanola.
Applications Claiming Priority (10)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201562159899P | 2015-05-11 | 2015-05-11 | |
| US201562183074P | 2015-06-22 | 2015-06-22 | |
| US201562185921P | 2015-06-29 | 2015-06-29 | |
| US201562203052P | 2015-08-10 | 2015-08-10 | |
| US201562220366P | 2015-09-18 | 2015-09-18 | |
| US201562222388P | 2015-09-23 | 2015-09-23 | |
| US201562236506P | 2015-10-02 | 2015-10-02 | |
| US201662313491P | 2016-03-25 | 2016-03-25 | |
| PCT/US2016/031797 WO2016183163A1 (en) | 2015-05-11 | 2016-05-11 | Expression constructs and methods of genetically engineering methylotrophic yeast |
| EP16793420.7A EP3294762B1 (en) | 2015-05-11 | 2016-05-11 | Expression constructs and methods of genetically engineering methylotrophic yeast |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| HRP20220299T1 true HRP20220299T1 (hr) | 2022-05-13 |
Family
ID=57249545
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| HRP20220299TT HRP20220299T1 (hr) | 2015-05-11 | 2016-05-11 | Ekspresijski konstrukti i postupci za genetsku modifikaciju metilotrofnog kvasca |
Country Status (22)
| Country | Link |
|---|---|
| US (6) | US10689656B2 (hr) |
| EP (2) | EP3294762B1 (hr) |
| JP (5) | JP6831797B2 (hr) |
| KR (4) | KR102229968B1 (hr) |
| CN (1) | CN107614516A (hr) |
| AU (4) | AU2016262494B2 (hr) |
| BR (1) | BR112017024123A2 (hr) |
| CA (1) | CA2985641A1 (hr) |
| CY (1) | CY1125979T1 (hr) |
| DK (1) | DK3294762T3 (hr) |
| ES (1) | ES2908463T3 (hr) |
| HR (1) | HRP20220299T1 (hr) |
| HU (1) | HUE058104T2 (hr) |
| LT (1) | LT3294762T (hr) |
| MX (3) | MX2017014469A (hr) |
| NZ (3) | NZ775948A (hr) |
| PL (1) | PL3294762T3 (hr) |
| PT (1) | PT3294762T (hr) |
| RS (1) | RS63099B1 (hr) |
| SI (1) | SI3294762T1 (hr) |
| SM (1) | SMT202200124T1 (hr) |
| WO (1) | WO2016183163A1 (hr) |
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