RU2560437C2 - Способ получения полимеров из белков шелка пауков - Google Patents
Способ получения полимеров из белков шелка пауков Download PDFInfo
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- RU2560437C2 RU2560437C2 RU2011145429/10A RU2011145429A RU2560437C2 RU 2560437 C2 RU2560437 C2 RU 2560437C2 RU 2011145429/10 A RU2011145429/10 A RU 2011145429/10A RU 2011145429 A RU2011145429 A RU 2011145429A RU 2560437 C2 RU2560437 C2 RU 2560437C2
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- Prior art keywords
- amino acid
- spider silk
- fragment
- acid residues
- silk protein
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
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- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
- C07K14/43513—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from arachnidae
- C07K14/43518—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from arachnidae from spiders
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Applications Claiming Priority (7)
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| CN102844437B (zh) | 2010-03-18 | 2015-06-03 | 思百博技术股份公司 | 蛋白质和多肽的制备 |
| CN103270049B (zh) | 2010-10-27 | 2017-10-13 | 思百博技术股份公司 | 用于结合有机靶的蜘蛛丝融合蛋白结构 |
| US9617315B2 (en) | 2011-06-01 | 2017-04-11 | Spiber Inc. | Artificial polypeptide fiber and method for producing the same |
| WO2013164405A1 (en) * | 2012-05-02 | 2013-11-07 | Spiber Technologies Ab | Spider silk fusion protein structures without repetitive fragment for binding to an organic target |
| EP3973992A3 (en) | 2013-09-17 | 2022-08-31 | Bolt Threads, Inc. | Methods and compositions for synthesizing improved silk fibers |
| AU2015358537B2 (en) | 2014-12-02 | 2021-08-19 | Evolved By Nature, Inc. | Silk performance apparel and products and methods of preparing the same |
| CN107709571B (zh) * | 2015-03-16 | 2021-11-02 | 保尔特纺织品公司 | 改善的丝纤维 |
| CN107735406A (zh) | 2015-04-09 | 2018-02-23 | 丝芭博株式会社 | 极性溶剂溶液及其制造方法 |
| EP3281948B1 (en) | 2015-04-09 | 2020-06-10 | Spiber Inc. | Polar solvent solution and production method thereof |
| JP2018521239A (ja) | 2015-06-11 | 2018-08-02 | ボルト スレッズ インコーポレイテッド | 改善された特性を有する組換えタンパク質繊維糸 |
| CN105031723B (zh) * | 2015-06-23 | 2017-05-24 | 上海交通大学 | 基于蜘蛛丝蛋白的温敏性水凝胶 |
| BR112018000699B1 (pt) | 2015-07-14 | 2023-10-17 | Silk Therapeutics, Inc | Artigo e método para revestir um material |
| JP2018159137A (ja) | 2015-08-20 | 2018-10-11 | 国立研究開発法人理化学研究所 | クモ糸様構造を有するポリペプチド繊維の製造方法 |
| EP3374378B1 (en) | 2015-11-13 | 2021-02-17 | Spiber Technologies AB | Charge-reversed n-terminal spider silk protein domain and uses thereof |
| GB201522610D0 (en) * | 2015-12-22 | 2016-02-03 | Univ Southampton | Protein |
| RU2018144307A (ru) * | 2016-05-16 | 2020-06-17 | Спайбер Текнолоджиз Аб | Покрытие из шелка паука для твердых поверхностей |
| EP3469121A4 (en) * | 2016-06-10 | 2020-01-08 | Bolt Threads, Inc. | RECOMBINANT PROTEIN FIBER YARNS WITH IMPROVED PROPERTIES |
| EP3263593A1 (en) | 2016-07-01 | 2018-01-03 | Anna Rising | Engineered spider silk proteins and uses thereof |
| CA3035839A1 (en) | 2016-09-14 | 2018-03-22 | Bolt Threads, Inc. | Long uniform recombinant protein fibers |
| EP4202394A1 (en) * | 2016-11-11 | 2023-06-28 | AMSilk GmbH | Use of a shrinkable biopolymer fiber as sensor |
| CN107475807A (zh) * | 2017-08-30 | 2017-12-15 | 常州豫春化工有限公司 | 一种改性尼龙6纤维的制备方法 |
| EP3688017A1 (en) * | 2017-09-26 | 2020-08-05 | Spiber Technologies AB | Structuring of surface-active macromolecules |
| EP3688018A4 (en) | 2017-09-27 | 2021-07-07 | Evolved by Nature, Inc. | SILK COVERED FABRICS, RELATED PRODUCTS AND PREPARATION PROCESSES |
| JP2021035330A (ja) | 2017-09-29 | 2021-03-04 | Ｓｐｉｂｅｒ株式会社 | 発現カセット |
| KR20220029660A (ko) * | 2019-07-04 | 2022-03-08 | 시비스 머테리얼 사이언스 리미티드 | 원핵생물 발현 시스템 및 이의 사용 방법 |
| CN110551194B (zh) * | 2019-09-19 | 2023-03-21 | 天津大学 | 重组蜘蛛卵鞘丝蛋白复合物及其生成的人造卵鞘丝 |
| AU2021259692A1 (en) | 2020-04-23 | 2022-10-27 | Seevix Material Sciences Ltd. | Cosmetic compositions comprising dragline spider silk |
| TW202432573A (zh) * | 2022-10-28 | 2024-08-16 | 國立臺灣大學 | 重組蜘蛛絲蛋白及其無標籤省時純化方法 |
| CN218413490U (zh) | 2022-10-31 | 2023-01-31 | 徐州龙安电子科技有限公司 | 一种新型蜘蛛电子标签 |
| DE102023105437B3 (de) | 2023-03-06 | 2024-06-13 | Dr. Ing. H.C. F. Porsche Aktiengesellschaft | Fahrzeugkomponente mit wenigstens einer Fahrzeuginnenraumkomponente |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007078239A2 (en) * | 2005-12-30 | 2007-07-12 | Spiber Technologies Ab | Spider silk proteins and methods for producing spider silk proteins |
| RU2007102846A (ru) * | 2004-07-22 | 2008-08-27 | Текнише Универзитет Мюнхен (DE) | Рекомбинантные белки шелка пауков |
| WO2008154547A2 (en) * | 2007-06-11 | 2008-12-18 | The Regents Of The University Of California | Spider silk dragline polynucleotides, polypeptides and methods of use thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2003201513A1 (en) * | 2002-01-11 | 2003-07-24 | Nexia Biotechnologies, Inc. | Methods of producing silk polypeptides and products thereof |
| BRPI0701826B1 (pt) * | 2007-03-16 | 2021-02-17 | Embrapa - Empresa Brasileira De Pesquisa Agropecuária | proteínas da teia de aranha nephilengys cruentata, avicularia juruensis e parawixia bistriata isoladas da biodiversidade brasileira |
-
2010
- 2010-04-21 RU RU2011145429/10A patent/RU2560437C2/ru active
- 2010-04-21 AU AU2010239773A patent/AU2010239773B2/en not_active Ceased
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- 2010-04-21 JP JP2012507182A patent/JP5912077B2/ja active Active
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2007102846A (ru) * | 2004-07-22 | 2008-08-27 | Текнише Универзитет Мюнхен (DE) | Рекомбинантные белки шелка пауков |
| WO2007078239A2 (en) * | 2005-12-30 | 2007-07-12 | Spiber Technologies Ab | Spider silk proteins and methods for producing spider silk proteins |
| WO2008154547A2 (en) * | 2007-06-11 | 2008-12-18 | The Regents Of The University Of California | Spider silk dragline polynucleotides, polypeptides and methods of use thereof |
Non-Patent Citations (2)
| Title |
|---|
| DB "UniprotKB", N Q05H60_9ARAC, 14.11.2006. DB "EMBL-EBI", N * |
| HEDHAMMAR M. et al, Structural properties of recombinant nonrepetitive and repetitive parts of major ampullate spidroin 1 from Euprosthenops australis: implications for fiber formation, Biochemistry, 2008, v. 47, p. 3407-3417. * |
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| EP2421890B1 (en) | 2015-08-26 |
| AU2010239773B2 (en) | 2014-11-06 |
| CN102395601A (zh) | 2012-03-28 |
| EP2421890A1 (en) | 2012-02-29 |
| JP2012531889A (ja) | 2012-12-13 |
| RU2011145429A (ru) | 2013-05-27 |
| DK2421890T3 (en) | 2015-11-23 |
| JP5912077B2 (ja) | 2016-04-27 |
| US8642734B2 (en) | 2014-02-04 |
| US20120041177A1 (en) | 2012-02-16 |
| WO2010123450A1 (en) | 2010-10-28 |
| EP2421890A4 (en) | 2012-10-24 |
| CA2759465A1 (en) | 2010-10-28 |
| AU2010239773A1 (en) | 2011-11-03 |
| CA2759465C (en) | 2017-12-12 |
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