DK3044591T3 - Rekombinante mikroorganismer og fremgangsmåder til anvendelse deraf - Google Patents
Rekombinante mikroorganismer og fremgangsmåder til anvendelse deraf Download PDFInfo
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- DK3044591T3 DK3044591T3 DK14844036.5T DK14844036T DK3044591T3 DK 3044591 T3 DK3044591 T3 DK 3044591T3 DK 14844036 T DK14844036 T DK 14844036T DK 3044591 T3 DK3044591 T3 DK 3044591T3
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- DK
- Denmark
- Prior art keywords
- clostridium
- phes
- seq
- nucleic acid
- phenylalanine
- 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/93—Ligases (6)
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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
- C12N15/74—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y601/00—Ligases forming carbon-oxygen bonds (6.1)
- C12Y601/01—Ligases forming aminoacyl-tRNA and related compounds (6.1.1)
- C12Y601/0102—Phenylalanine-tRNA ligase (6.1.1.20)
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- Proteomics, Peptides & Aminoacids (AREA)
- Toxicology (AREA)
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
Claims (15)
1. Modificeret phenylalanin-tRNA-syntetase (PheS), hvor den modificerede PheS har sekvensen ifølge SEQ ID No. 21, som omfatter mindst en ændring i aminosyrepositioneme 306 til 313.
2. Modificeret PheS ifølge krav 1, hvor den modificerede PheS aminoacylerer tRNA med en phenylalaninanalog, der er valgt fra gruppen bestående af chlorphenylalanin, fluorphenylalanin og bromphenylalanin.
3. Modificeret PheS ifølge krav 2, hvor phenylalaninanalogen er valgt fra gruppen bestående af DL-4-chlorphenylalanin, p-chlorphenylalanin, p-fluor-L-phenylalanin, p-fluor-DL-phenylalanin og p-brom-L-phenylalanin.
4. Modificeret PheS ifølge krav 1, hvor den modificerede PheS omfatter en ændring i aminosyrepositionen 311 i SEQ ID No. 21.
5. Modificeret PheS ifølge krav 4, hvor den modificerede PheS omfatter glycin i stedet for alanin i aminosyrepositionen 311 i SEQ ID No. 21.
6. Nukleinsyre, der koder for den modificerede PheS ifølge krav 1.
7. Fremgangsmåde til fremstilling af en rekombinant mikroorganisme, hvilken fremgangsmåde omfatter at indføre en vektor, som omfatter en nukleinsyre, der koder for den modificerede PheS ifølge krav 1, i en forældremikroorganisme.
8. Fremgangsmåde ifølge krav 7, hvor fremgangsmåden endvidere omfatter selektion i forhold til ekspression af den modificerede PheS.
9. Fremgangsmåde ifølge krav 8, hvor selektionen i forhold til ekspression af den modificerede PheS indebærer at dyrke den rekombinante mikroorganisme i nærvær af en phenylalaninanalog, der er valgt fra gruppen bestående af chlorphenylalanin, fluorphenylalanin og bromphenylalanin.
10. Fremgangsmåde ifølge krav 9, hvor phenylalaninanalogen er valgt fra gruppen bestående af DL-4-chlorphenylalanin, p-chlorphenylalanin, p-fluor-L-phenylalanin, p-fluor-DL-phenylalanin og p-brom-L-phenylalanin.
11. Fremgangsmåde ifølge krav 7, hvor vektoren endvidere omfatter homologiarme, der tillader homolog rekombination mellem vektoren og forældremikroorganismens genom, en nukleinsyre af interesse og/eller en positiv selektionsmarkør.
12. Fremgangsmåde ifølge krav 7, hvor fremgangsmåden endvidere omfatter selektion for ekspression af den positive selektionsmarkør, fortrinsvis hvor den positive selektionsmarkør er valgt fra gruppen bestående af CatP, ErmB og TetA.
13. Anvendelse af PheS ifølge krav 1 som en modselektionsmarkør.
14. Fremgangsmåde ifølge krav 9, hvor den modificerede PheS omfatter en ændring i aminosyrepositionen 311 i SEQ ID No. 21.
15. Fremgangsmåde ifølge krav 14, hvor den modificerede PheS omfatter glycin i stedet for alanin i aminosyrepositionen 311 i SEQ ID No. 21.
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| CN114507695B (zh) * | 2022-02-17 | 2023-09-19 | 四川大学华西医院 | 一种用于调控神经元活性的重组载体和方法 |
| CN115637242B (zh) * | 2022-10-20 | 2024-09-27 | 中南大学 | 微生物复合菌剂及其制备方法和赤泥原位生物脱碱的方法 |
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| US7704723B2 (en) | 2006-08-31 | 2010-04-27 | The Board Of Regents For Oklahoma State University | Isolation and characterization of novel clostridial species |
| CN101528930B (zh) * | 2006-10-03 | 2013-12-11 | 代谢探索者公司 | 用于在梭菌属中进行染色体整合和dna序列替换的方法 |
| EP3176261A1 (en) * | 2008-06-26 | 2017-06-07 | aTyr Pharma, Inc. | Compositions and methods comprising glycyl-trna synthetases having non-canonical biological activities |
| CN102348807B (zh) * | 2009-01-12 | 2015-04-15 | 苏特罗生物制药公司 | 利用体外蛋白合成体系将非天然氨基酸引入蛋白的单装载体系 |
| CA2776757A1 (en) * | 2009-08-10 | 2011-02-17 | Shital Tripathi | Positive and negative selectable markers for use in thermophilic organisms |
| WO2011101696A1 (en) * | 2010-02-18 | 2011-08-25 | Cellectis | Improved meganuclease recombination system |
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| CN102181516A (zh) * | 2011-02-18 | 2011-09-14 | 中国医学科学院医药生物技术研究所 | 一种PheRS酶活测定体系以及PheRS抑制剂的筛选方法 |
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| CN105723219A (zh) | 2016-06-29 |
| CN105723219B (zh) | 2018-06-05 |
| AU2014318672A1 (en) | 2016-03-03 |
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| US9745566B2 (en) | 2017-08-29 |
| EP3044591A1 (en) | 2016-07-20 |
| WO2015038853A1 (en) | 2015-03-19 |
| EP3044591B1 (en) | 2018-08-29 |
| AU2014318672B2 (en) | 2020-01-30 |
| CA2921430C (en) | 2018-04-24 |
| CA2921430A1 (en) | 2015-03-19 |
| US20150072395A1 (en) | 2015-03-12 |
| SG11201601263WA (en) | 2016-03-30 |
| KR20160043983A (ko) | 2016-04-22 |
| KR102282778B1 (ko) | 2021-07-30 |
| EP3044591A4 (en) | 2017-02-22 |
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