DK2576796T3 - Filamentøse svampeværtsstammer og dna-konstrukter samt fremgangsmåder til anvendelse deraf - Google Patents
Filamentøse svampeværtsstammer og dna-konstrukter samt fremgangsmåder til anvendelse deraf Download PDFInfo
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- DK2576796T3 DK2576796T3 DK11727041.3T DK11727041T DK2576796T3 DK 2576796 T3 DK2576796 T3 DK 2576796T3 DK 11727041 T DK11727041 T DK 11727041T DK 2576796 T3 DK2576796 T3 DK 2576796T3
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- 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
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Claims (12)
- FILAMENTØSE SVAMPEVÆRTSSTAMMER OG DNA-KONSTRUKTER SAMT FREMGANGSMÅDER TIL ANVENDELSE DERAF1. Eilamentøs svampe værtscelle-ekspressionssystem, der omfatter: a. en svampeværtscelle, der i sit kromosomale DNA indeholder en disruption i én eller flere komponenter af den ikke-homologe rekombinations- (NHR) bane, en del af en første valgbar markør, der mangler en første valgbar funktion, og en anden valgbar markør, der kan anvendes til at tildele en anden valgbar funktion; og b. et nukleinsyremolekyle indeholdende en sekvens, der, når den introduceres i svampeværtscellen, tildeler den første valgbare markør den første valgbare funktion, en sekvens, der kan anvendes til at udtrykke ét eller flere gener af interesse, og sekvenser med væsentlig homologi med sekvenser, der flankerer de kromosomale valgbare markører; hvor de homologe sekvenser bevirker et homologt rekombinations-event, der resulterer i en første funktionel valgbar markør, fjernelse af den anden valgbare markør og ekspression af genet af interesse, og hvor den første valgbare markør og den anden valgbare markør er forskellige markører.
- 2. Filamentøs svampeværtscelle-ekspressionssystem ifølge krav 1, hvor den ene eller flere komponenter af NHR-banen er udvalgt fra gruppen bestående af ku80, ku70, rad50, mrell, xrs2, lig4 og xrs.
- 3. Filamentøs svampeværtscelle-ekspressionssystem ifølge krav 1, hvor den første valgbare markør og den anden valgbare markør er forskellige markører udvalgt fra gruppen bestående af alsR, amdS, hygR, pyr2, pyr4, pyrG, sucA, en bleomycinresistensmarkør, en blasticidinresistensmarkør, en pyrithiaminresistensmarkør, en chlorimuronethylresistensmarkør, en neomycinresistensmarkør, et adeninbanegen, et tryptophanbanegen og thymidinkinase.
- 4. Filamentøs svampeværtscelle-ekspressionssystem ifølge krav 1, hvor svampeværtscellen er fra en art, der er udvalgt fra gruppen bestående af Trichoderma, Penicillium, Aspergillus, Humicola, Chrysosporium, Fusarium, Neurospora og Emericella.
- 5. Filamentøs svampeværtscelle-ekspressionssystem ifølge krav 4, hvor Trichoderma er T. reesei.
- 6. Filamentøs svampe værtscelle-ekspressionssystem ifølge krav 4, hvor Aspergillus er A. niger.
- 7. Filamentøs svampeværtscelle-ekspressionssystem ifølge krav 1, hvor genet af interesse er udvalgt fra gruppen bestående af hemicellulaser, peroxidaser, proteaser, cellulaser, xylanaser, lipaser, phospholipaser, esteraser, cutinaser, pectinaser, keratinaser, reduktaser, oxidaser, phenoloxidaser, lipoxygenaser, ligninaser, pullulanaser, tannaser, pentosanaser, malanaser, beta-glucanaser, arabinosidaser, hyaluronidase, chondroitinase, laccase, amylaser, glucoamylaser og blandinger deraf.
- 8. Filamentøs svampeværtscelle-ekspressionssystem ifølge krav 1, hvor genet af interesse er udvalgt fra gruppen bestående af acetylesteraser, aminopeptidaser, amylaser, arabinaser, arabinofuranosidaser, carboxypeptidaser, catalaser, cellulaser, chitinaser, chymosin, cutinase, deoxyribonucleaser, epimeraser, esteraser, a-galactosidaser, β-galac tos idaser, a-glucanaser, glucanlysaser, endo^-glucanaser, glucoamylaser, glucoseoxidaser, a-glucosidaser, β-glucosidaser, glucuronidaser, hemicellulaser, hexoseoxidaser, hydrolaser, invertaser, isomeraser, laccaser, lipaser, lyaser, mannosidaser, oxidaser, oxidoreduktaser, pectatlyaser, pectinacetylresteraser, pectinrdepolymeraser, pectinmethylesteraser, pectinolytiske enzymer, peroxidaser, phenoloxidaser, phytaser, polygalacturonaser, proteaser, rhamno-galacturonaser, ribonucleaser, thaumatin, transferaser, transportproteiner, transglutaminaser, xylanaser, hexoseoxidaser og kombinationer deraf.
- 9. Filamentøs svampeværtscelle-ekspressionssystem ifølge krav 1, hvor genet af interesse er udvalgt fra gruppen bestående af peptidhormoner, vækstfaktorer, koaguleringsfaktorer, chemokiner, cytokiner, lymfokiner, antistoffer, receptorer, adhæsionsmolekyler, mikrobielle antigener og fragmenter deraf.
- 10. Fremgangsmåde til ekspression af et gen af interesse i det filamentøse svampeværtscellesystem ifølge et hvilket som helst af kravene 1 til 9, hvilken fremgangsmåde omfatter introduktion i den filamentøse svampeværtscelle af nukleinsyremolekylet, dyrkning af værtscellerne og udvælgelse af de værtsceller, der har den første valgbare funktion, men mangler den anden valgbare funktion.
- 11. Fremgangsmåde ifølge krav 10, hvilken fremgangsmåde endvidere omfatter udførelse af assay for ekspression af genet af interesse.
- 12. Fremgangsmåde ifølge krav 11, hvilken fremgangsmåde endvidere omfatter udførelse af assay for en biokemisk funktion af ct polypeptid, der cr kodet for af genet af interesse.
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| CN105992821A (zh) * | 2013-12-03 | 2016-10-05 | 诺维信公司 | 真菌基因文库的双分裂标记物整合 |
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| CN107109424A (zh) * | 2014-12-01 | 2017-08-29 | 丹尼斯科美国公司 | 真菌宿主菌株、dna构建体及使用方法 |
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| DK3256598T3 (da) * | 2015-02-09 | 2021-01-11 | Danisco Us Inc | Svampestammer og anvendelsesfremgangsmåder |
| CN104894165B (zh) * | 2015-05-26 | 2019-01-25 | 中国科学院青岛生物能源与过程研究所 | 一种提高基因打靶技术在土曲霉中应用效率的方法与应用 |
| CN104894115B (zh) * | 2015-05-26 | 2018-11-13 | 中国科学院青岛生物能源与过程研究所 | 一种具有高效同源重组能力的土曲霉及其构建方法与应用 |
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| EP3239898A1 (en) * | 2016-04-27 | 2017-11-01 | Mercarista, S.L. | Method for quantifying proteins in a sample |
| EP3478845A4 (en) | 2016-06-30 | 2019-07-31 | Zymergen, Inc. | METHOD OF GENERATING A GLUCOSE PERMEASE LIBRARY AND USES THEREOF |
| US10544390B2 (en) | 2016-06-30 | 2020-01-28 | Zymergen Inc. | Methods for generating a bacterial hemoglobin library and uses thereof |
| US10876103B2 (en) * | 2016-10-04 | 2020-12-29 | Danisco Us Inc | Protein production in filamentous fungal cells in the absence of inducing substrates |
| EP3562945A4 (en) * | 2016-12-30 | 2020-08-05 | Zymergen, Inc. | PROCESS FOR CONSTRUCTING FUNGAL PRODUCTION STRAINS USING AUTOMATED STEPS FOR GENETIC MANIPULATION AND STRAIN PURIFICATION |
| CN108588060B (zh) * | 2017-03-07 | 2020-12-01 | 武汉康复得生物科技股份有限公司 | 一种用丝状真菌宿主细胞表达的重组草酸脱羧酶 |
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| CN108004153B (zh) * | 2017-12-07 | 2021-05-28 | 潍坊康地恩生物科技有限公司 | 一株高产果胶裂解酶的里氏木霉菌株及其应用 |
| WO2019236848A1 (en) | 2018-06-06 | 2019-12-12 | Zymergen Inc. | Manipulation of genes involved in signal transduction to control fungal morphology during fermentation and production |
| CN114391038A (zh) * | 2019-07-25 | 2022-04-22 | 诺维信公司 | 丝状真菌表达系统 |
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| EP2576796B1 (en) | 2017-03-29 |
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| WO2011153449A1 (en) | 2011-12-08 |
| JP2013533737A (ja) | 2013-08-29 |
| CN102939382B (zh) | 2017-02-15 |
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