WO2024055291A1 - Procédé de criblage à haut débit de mutant de sirtuine, mutant de sirtuine et son utilisation - Google Patents
Procédé de criblage à haut débit de mutant de sirtuine, mutant de sirtuine et son utilisation Download PDFInfo
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- WO2024055291A1 WO2024055291A1 PCT/CN2022/119322 CN2022119322W WO2024055291A1 WO 2024055291 A1 WO2024055291 A1 WO 2024055291A1 CN 2022119322 W CN2022119322 W CN 2022119322W WO 2024055291 A1 WO2024055291 A1 WO 2024055291A1
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- Prior art keywords
- sirtuin
- mutant
- amc
- lysis
- activity
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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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
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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
- C12N15/01—Preparation of mutants without inserting foreign genetic material therein; Screening processes therefor
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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.)
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Abstract
La présente invention concerne un mutant de sirtuine, un procédé de criblage à haut rendement d'un mutant de sirtuine, et l'utilisation. Le procédé de criblage d'un mutant de sirtuine comprend les étapes suivantes : construction d'une banque de mutants de sirtuine à l'aide d'une technologie PCR à risque d'erreur; lyse de chaque mutant de la banque de mutants de sirtuine, réalisation d'une séparation solide-liquide et collecte d'un surnageant de lysat; et détermination de l'activité de désacylation de la sirtuine dans le surnageant de lysat à l'aide d'un procédé de fluorescence AMC, et obtention d'une souche de mutant de sirtuine présentant l'activité de désacylation requise. En utilisant le procédé de criblage pour un mutant de sirtuine, un mutant de l'enzyme de désacylation sirtuine peut être sélectionné de manière simple, pratique et répétée à l'aide d'un procédé à haut débit.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/CN2022/119322 WO2024055291A1 (fr) | 2022-09-16 | 2022-09-16 | Procédé de criblage à haut débit de mutant de sirtuine, mutant de sirtuine et son utilisation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/CN2022/119322 WO2024055291A1 (fr) | 2022-09-16 | 2022-09-16 | Procédé de criblage à haut débit de mutant de sirtuine, mutant de sirtuine et son utilisation |
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| Publication Number | Publication Date |
|---|---|
| WO2024055291A1 true WO2024055291A1 (fr) | 2024-03-21 |
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| Application Number | Title | Priority Date | Filing Date |
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| PCT/CN2022/119322 WO2024055291A1 (fr) | 2022-09-16 | 2022-09-16 | Procédé de criblage à haut débit de mutant de sirtuine, mutant de sirtuine et son utilisation |
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| Country | Link |
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| WO (1) | WO2024055291A1 (fr) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006065076A1 (fr) * | 2004-12-14 | 2006-06-22 | Cj Corporation | Mutant de la 5'-xmp aminase |
| CN103097545A (zh) * | 2010-07-07 | 2013-05-08 | 康奈尔大学 | Sirt5调节剂及其筛选方法 |
| CN107630011A (zh) * | 2006-07-05 | 2018-01-26 | 催化剂生物科学公司 | 蛋白酶筛选方法及由此鉴别的蛋白酶 |
| WO2018064752A1 (fr) * | 2016-10-07 | 2018-04-12 | Ranomics Inc. | Procédé basé sur la pcr pour produire des banques d'adn mutagènes de saturation multisites |
| CN108192882A (zh) * | 2018-01-18 | 2018-06-22 | 天津市湖滨盘古基因科学发展有限公司 | 一种人的组蛋白去乙酰化酶突变蛋白及其应用 |
| CN113584016A (zh) * | 2021-08-17 | 2021-11-02 | 安徽金禾实业股份有限公司 | 一种通过易错pcr技术及高通量筛选提高葡萄糖基转移酶eugt11酶活方法 |
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2022
- 2022-09-16 WO PCT/CN2022/119322 patent/WO2024055291A1/fr unknown
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