JP2004532033A - 発現し難いタンパク質用の迅速に開裂可能なｓｕｍｏ融合タンパク質発現系 - Google Patents
発現し難いタンパク質用の迅速に開裂可能なｓｕｍｏ融合タンパク質発現系 Download PDFInfo
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- JP2004532033A JP2004532033A JP2002587558A JP2002587558A JP2004532033A JP 2004532033 A JP2004532033 A JP 2004532033A JP 2002587558 A JP2002587558 A JP 2002587558A JP 2002587558 A JP2002587558 A JP 2002587558A JP 2004532033 A JP2004532033 A JP 2004532033A
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- ulp1
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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
- 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
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/62—DNA sequences coding for fusion proteins
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
- C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
- C07K2319/21—Fusion polypeptide containing a tag with affinity for a non-protein ligand containing a His-tag
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
- C07K2319/23—Fusion polypeptide containing a tag with affinity for a non-protein ligand containing a GST-tag
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/35—Fusion polypeptide containing a fusion for enhanced stability/folding during expression, e.g. fusions with chaperones or thioredoxin
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/40—Fusion polypeptide containing a tag for immunodetection, or an epitope for immunisation
- C07K2319/43—Fusion polypeptide containing a tag for immunodetection, or an epitope for immunisation containing a FLAG-tag
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/95—Fusion polypeptide containing a motif/fusion for degradation (ubiquitin fusions, PEST sequence)
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- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US28865601P | 2001-05-04 | 2001-05-04 | |
| US32908001P | 2001-10-12 | 2001-10-12 | |
| PCT/US2002/014062 WO2002090495A2 (en) | 2001-05-04 | 2002-05-06 | Rapidly cleavable sumo fusion protein expression system for difficult to express proteins |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2004532033A true JP2004532033A (ja) | 2004-10-21 |
| JP2004532033A5 JP2004532033A5 (enExample) | 2010-02-25 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2002587558A Pending JP2004532033A (ja) | 2001-05-04 | 2002-05-06 | 発現し難いタンパク質用の迅速に開裂可能なｓｕｍｏ融合タンパク質発現系 |
Country Status (8)
| Country | Link |
|---|---|
| US (3) | US6872551B2 (enExample) |
| EP (1) | EP1392717B1 (enExample) |
| JP (1) | JP2004532033A (enExample) |
| AT (1) | ATE490968T1 (enExample) |
| AU (1) | AU2002257243A1 (enExample) |
| CA (1) | CA2445049A1 (enExample) |
| DE (1) | DE60238532D1 (enExample) |
| WO (1) | WO2002090495A2 (enExample) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2005514025A (ja) * | 2002-01-07 | 2005-05-19 | ライフセンサーズ、インク． | タンパク質発現及び精製のための方法及び組成 |
| JP2018531017A (ja) * | 2015-10-14 | 2018-10-25 | トランスレイト バイオ， インコーポレイテッド | 生産性向上のためのｒｎａ関連酵素の修飾 |
Families Citing this family (49)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001009292A2 (en) * | 1999-07-31 | 2001-02-08 | Board Of Regents, The University Of Texas System | Sentrin-specific human proteases senp1-3 |
| US7198924B2 (en) | 2000-12-11 | 2007-04-03 | Invitrogen Corporation | Methods and compositions for synthesis of nucleic acid molecules using multiple recognition sites |
| DE60238532D1 (de) * | 2001-05-04 | 2011-01-20 | Cornell Res Foundation Inc | Schnell spaltbares sumofusionsprotein-expressionssystem für schwer zu exprimierende proteine |
| US8535908B2 (en) * | 2002-01-04 | 2013-09-17 | Brookhaven Science Associates, Llc | Facilitating protein solubility by use of peptide extensions |
| US7220576B2 (en) * | 2002-01-07 | 2007-05-22 | Lifesensors, Inc. | Methods and compositions for protein expression and purification |
| JP2005532829A (ja) * | 2002-07-18 | 2005-11-04 | インヴィトロジェン コーポレーション | 組換え部位を含むウイルスベクター |
| WO2004063214A2 (en) * | 2003-01-08 | 2004-07-29 | Government Of The United States Of America As Represented By The Sercretary Of The Department Of Health And Human Services National Institutes Of Health | Antibodies that specifically recognize sumo-conjugated proteins |
| CA2530530A1 (en) * | 2003-06-26 | 2005-01-13 | Lifesensors, Inc. | Methods and compositions for enhanced protein expression and purification |
| WO2005014796A2 (en) | 2003-08-08 | 2005-02-17 | Invitrogen Corporation | Methods and compositions for seamless cloning of nucleic acid molecules |
| US8507277B2 (en) | 2003-10-24 | 2013-08-13 | Gencia Corporation | Nonviral vectors for delivering polynucleotides |
| US8133733B2 (en) | 2003-10-24 | 2012-03-13 | Gencia Corporation | Nonviral vectors for delivering polynucleotides to target tissues |
| WO2005056752A2 (en) | 2003-10-24 | 2005-06-23 | Gencia Corporation | Methods and compositions for delivering polynucleotides |
| US8062891B2 (en) | 2003-10-24 | 2011-11-22 | Gencia Corporation | Nonviral vectors for delivering polynucleotides to plants |
| US20090208478A1 (en) * | 2003-10-24 | 2009-08-20 | Gencia Corporation | Transducible polypeptides for modifying metabolism |
| US20090123468A1 (en) | 2003-10-24 | 2009-05-14 | Gencia Corporation | Transducible polypeptides for modifying metabolism |
| DE602004027538D1 (de) | 2003-12-01 | 2010-07-15 | Life Technologies Corp | Rekombinationsstellen enthaltende nukleinsäuremoleküle und verfahren zur verwendung davon |
| ES2306541B1 (es) * | 2004-04-16 | 2009-09-28 | Consejo Superior Investig. Cientificas | Trasplante funcional de la maquinaria de sumoilacion a escherichia coli. |
| CA2576083A1 (en) * | 2004-08-18 | 2006-02-23 | Joel Weiner | Protein production method utilizing yebf |
| US8119369B2 (en) * | 2004-12-30 | 2012-02-21 | Lifesensors, Inc. | Human SUMO-3 for enhancing protein expression |
| CA2670014A1 (en) * | 2006-11-22 | 2008-11-20 | Emory University | Production of anti-microbial peptides |
| EP2100961A1 (en) * | 2008-03-04 | 2009-09-16 | Icon Genetics GmbH | Method of protease production in plants |
| US20090280531A1 (en) * | 2008-05-06 | 2009-11-12 | Academia Sinica | Preparation of Soluble Capsid Proteins of Picornaviruses Using SUMO Fusion Technology |
| TW200951219A (en) * | 2008-05-06 | 2009-12-16 | Academia Sinica | An improved SUMO fusion protein system for effective production of native proteins |
| US8440449B2 (en) * | 2008-09-30 | 2013-05-14 | The United States Of America, As Represented By The Secretary Of Agriculture | Transformed Saccharomyces cerevisiae engineered for xylose utilization |
| US8663951B2 (en) * | 2009-01-09 | 2014-03-04 | Academia Sinica | Method of producing virus-like particles of picornavirus using a small-ubiquitin-related fusion protein expression system |
| CN102533712A (zh) * | 2009-05-12 | 2012-07-04 | 中国科学院上海生命科学研究院 | 一种固相化sumo化系统及固相化去sumo化系统 |
| CN101993495B (zh) * | 2009-08-12 | 2013-07-24 | 上海近岸科技有限公司 | 一种蛋白质混合物及其制备方法 |
| CN103184228A (zh) * | 2011-12-29 | 2013-07-03 | 天津市国际生物医药联合研究院 | 用于可结晶成熟型SARS冠状病毒非结构蛋白12(sars-nsp12)的外源重组表达及纯化方法 |
| WO2013103964A1 (en) | 2012-01-06 | 2013-07-11 | Gencia Corporation | Methods and compositions for treating proteinopathies |
| WO2013188873A1 (en) | 2012-06-15 | 2013-12-19 | Gencia Corporation | Compositions and methods for enhancing immune responses |
| US20160038565A1 (en) | 2012-06-15 | 2016-02-11 | Gencia Corporation | Methods of Mitigating Side Effects of Radiation Exposure and Chemotherapy |
| AU2013326968B2 (en) | 2012-10-03 | 2019-08-08 | Agrivida, Inc. | Multiprotein expression cassettes |
| KR101911438B1 (ko) | 2012-10-31 | 2018-10-24 | 삼성전자주식회사 | 이중 특이 항원 결합 단백질 복합체 및 이중 특이 항체의 제조 방법 |
| US9707244B2 (en) | 2013-03-15 | 2017-07-18 | Gencia Corporation | Compositions and methods for treating conditions that affect epidermis |
| US9970945B2 (en) | 2013-06-07 | 2018-05-15 | Georgia State University Research Foundtion, Inc. | Compositions, methods, and assays using proteo-liposome-transfected oocytes for analyzing the structure, function, and activity of membrane proteins |
| US9447157B2 (en) | 2013-09-13 | 2016-09-20 | Augusta University Research Institute, Inc. | Nitration shielding peptides and methods of use thereof |
| WO2015049230A1 (en) | 2013-10-01 | 2015-04-09 | MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. | An orthogonal set of tag-cleaving proteases for purification of proteins and stoichiometric protein complexes |
| US10378003B2 (en) | 2014-11-10 | 2019-08-13 | MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. | Tag removal from proteins expressed in pro- and eukaryotic hosts |
| TWI591177B (zh) * | 2016-10-27 | 2017-07-11 | 中化合成生技股份有限公司 | 一種胰高血糖素胜肽－２（ｇｌｐ－２）類似物的製備方法 |
| US10913777B2 (en) | 2017-05-22 | 2021-02-09 | The Regents Of The University Of California | In vitro assembly of bacterial microcompartments |
| US20190070262A1 (en) | 2017-09-06 | 2019-03-07 | Yale University | Interleukin-18 variants and methods of use |
| KR20190135393A (ko) * | 2018-05-28 | 2019-12-06 | 주식회사 원진바이오테크놀로지 | 폴리유비퀴틴 스캐폴드에 결합된 생체분자들의 선형 멀티머 중합체 및 이의 용도 |
| CN109517074A (zh) * | 2018-12-04 | 2019-03-26 | 湖北大学 | 一种融合标签促进Kuma030蛋白酶可溶性表达及非亲和层析快速纯化的应用 |
| CN109628479A (zh) * | 2018-12-25 | 2019-04-16 | 天津农学院 | 一种胰高血糖素样肽-1的重组表达方法 |
| EP3906248A4 (en) * | 2019-01-04 | 2022-08-10 | Repertoire Immune Medicines, Inc. | METHODS FOR PRODUCING HIGH DIVERSITY PEPTIDE BANKS AND PROMOTING PROTEIN FOLDING |
| CN113025675B (zh) * | 2021-05-21 | 2021-08-20 | 凯莱英医药集团（天津）股份有限公司 | 多肽的制备方法 |
| EP4479088A1 (en) | 2022-02-14 | 2024-12-25 | University of Georgia Research Foundation, Inc. | Pan-pneumovirus vaccine compositions and methods of use thereof |
| CN114426586A (zh) * | 2022-02-21 | 2022-05-03 | 中国农业大学 | 一种胞外分泌表达Ulp1蛋白酶的大肠杆菌基因工程菌株及其应用 |
| CN114539425B (zh) * | 2022-02-25 | 2023-04-28 | 湖南中晟全肽生化有限公司 | 一种提高线性多肽生物表达的方法 |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5132213A (en) * | 1986-10-02 | 1992-07-21 | Massachusetts Institute Of Technology | Method for producing proteins and polypeptides using ubiquitin fusions |
| US5643758A (en) * | 1987-03-10 | 1997-07-01 | New England Biolabs, Inc. | Production and purification of a protein fused to a binding protein |
| AU6290496A (en) * | 1995-06-27 | 1997-01-30 | Igen International, Inc. | High-level expression and efficient recovery of ubiquitin fusion proteins from escherichia coli |
| DE60238532D1 (de) * | 2001-05-04 | 2011-01-20 | Cornell Res Foundation Inc | Schnell spaltbares sumofusionsprotein-expressionssystem für schwer zu exprimierende proteine |
| US7060461B2 (en) * | 2002-01-07 | 2006-06-13 | Lifesensors, Inc. | Methods and compositions for protein expression and purification |
-
2002
- 2002-05-06 DE DE60238532T patent/DE60238532D1/de not_active Expired - Lifetime
- 2002-05-06 CA CA002445049A patent/CA2445049A1/en not_active Abandoned
- 2002-05-06 AU AU2002257243A patent/AU2002257243A1/en not_active Abandoned
- 2002-05-06 WO PCT/US2002/014062 patent/WO2002090495A2/en not_active Ceased
- 2002-05-06 JP JP2002587558A patent/JP2004532033A/ja active Pending
- 2002-05-06 AT AT02726837T patent/ATE490968T1/de not_active IP Right Cessation
- 2002-05-06 EP EP02726837A patent/EP1392717B1/en not_active Expired - Lifetime
- 2002-07-02 US US10/188,343 patent/US6872551B2/en not_active Expired - Lifetime
-
2004
- 2004-09-24 US US10/949,685 patent/US7498165B2/en not_active Expired - Fee Related
-
2008
- 2008-10-10 US US12/249,334 patent/US7910364B2/en not_active Expired - Lifetime
Non-Patent Citations (2)
| Title |
|---|
| JPN6008007552, Ｎａｔｕｒｅ，（１９９９），３９８，ｐ２４６−２５１ * |
| JPN6008007554, Ｍｏｌｅｃｕｌａｒ Ｃｅｌｌ，（２０００），５，ｐ８６５−８７６ * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| JP2005514025A (ja) * | 2002-01-07 | 2005-05-19 | ライフセンサーズ、インク． | タンパク質発現及び精製のための方法及び組成 |
| JP2018531017A (ja) * | 2015-10-14 | 2018-10-25 | トランスレイト バイオ， インコーポレイテッド | 生産性向上のためのｒｎａ関連酵素の修飾 |
| JP2021155447A (ja) * | 2015-10-14 | 2021-10-07 | トランスレイト バイオ， インコーポレイテッド | 生産性向上のためのｒｎａ関連酵素の修飾 |
| JP6997704B2 (ja) | 2015-10-14 | 2022-02-04 | トランスレイト バイオ， インコーポレイテッド | 生産性向上のためのｒｎａ関連酵素の修飾 |
| JP7119181B2 (ja) | 2015-10-14 | 2022-08-16 | トランスレイト バイオ， インコーポレイテッド | 生産性向上のためのｒｎａ関連酵素の修飾 |
| JP7119181B6 (ja) | 2015-10-14 | 2022-11-11 | トランスレイト バイオ， インコーポレイテッド | 生産性向上のためのｒｎａ関連酵素の修飾 |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2002090495A2 (en) | 2002-11-14 |
| WO2002090495A8 (en) | 2003-07-03 |
| US7910364B2 (en) | 2011-03-22 |
| CA2445049A1 (en) | 2002-11-14 |
| DE60238532D1 (de) | 2011-01-20 |
| US20050069988A1 (en) | 2005-03-31 |
| US7498165B2 (en) | 2009-03-03 |
| EP1392717B1 (en) | 2010-12-08 |
| EP1392717A4 (en) | 2005-06-08 |
| US20030086918A1 (en) | 2003-05-08 |
| US20090317868A1 (en) | 2009-12-24 |
| US6872551B2 (en) | 2005-03-29 |
| AU2002257243A1 (en) | 2002-11-18 |
| EP1392717A1 (en) | 2004-03-03 |
| ATE490968T1 (de) | 2010-12-15 |
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