JP5090304B2 - 突然変異体ルシフェラーゼ - Google Patents
突然変異体ルシフェラーゼ Download PDFInfo
- Publication number
- JP5090304B2 JP5090304B2 JP2008244507A JP2008244507A JP5090304B2 JP 5090304 B2 JP5090304 B2 JP 5090304B2 JP 2008244507 A JP2008244507 A JP 2008244507A JP 2008244507 A JP2008244507 A JP 2008244507A JP 5090304 B2 JP5090304 B2 JP 5090304B2
- Authority
- JP
- Japan
- Prior art keywords
- luciferase
- mutant
- luciferases
- beetle
- amino acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
Classifications
-
- 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
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0069—Oxidoreductases (1.) acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y113/00—Oxidoreductases acting on single donors with incorporation of molecular oxygen (oxygenases) (1.13)
- C12Y113/12—Oxidoreductases acting on single donors with incorporation of molecular oxygen (oxygenases) (1.13) with incorporation of one atom of oxygen (internal monooxygenases or internal mixed function oxidases)(1.13.12)
- C12Y113/12007—Photinus-luciferin 4-monooxygenase (ATP-hydrolysing) (1.13.12.7), i.e. firefly-luciferase
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Enzymes And Modification Thereof (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Luminescent Compositions (AREA)
Description
本発明は、甲虫類の突然変異体ルシフェラーゼ、及び該突然変異体ルシフェラーゼをコードするDNAを提供する。好ましくは、上記突然変異体ルシフェラーゼは対応する野生型ルシフェラーゼとは異なる色の発光を生起し、この色の相違は好ましくは、突然変異体の発光ピーク強度波長が野生型酵素のものと少なくとも1nm相違することによる。
本発明の突然変異体ルシフェラーゼは対応する野生型ルシフェラーゼから、1個以上の、しかし典型的には2個以下のアミノ酸の置換によって相違する。本発明のルシフェラーゼは、3個のカルボキシ末端アミノ酸のうちの1個以上を改変してペルオキシソーム標的化を低減することも可能である。
以下の説明及び実施例において方法実施及び濃度測定は、特に記載のない限りは室温(約20℃〜25℃)及び大気圧で行った。
ポリペプチド中のある位置での置換は、[元のアミノ酸の記号]位置番号[置換アミノ酸の記号]で表わされる。
例えば、ポリペプチドの位置100にあるアラニンがグルタミン酸で置換されていることは、Ala100GluまたはA100Eと表わされる。通常は、置換表示の前にそれが生じたポリペプチドを表示する。例えば、“Luck”と名付ける仮想タンパク質に置換A100Eが存在する場合、置換はLuck−Ala100GluまたはLuck−A100Eと表わされる。ポリペプチド中に2つ以上の置換がある場合、置換表示はスラッシュ(/)によって区切られる。例えば、仮想タンパク質“Luck”において、位置100のアラニンがグルタミン酸で置換され、位置150のリシンがアスパラギンで置換されている場合、置換を有するポリペプチドはLuck−Ala100Glu/Lys150AsnまたはLuckA100E/K150Nと表わされる。ポリペプチド中の1つの位置に異なる置換があることを示すためには、置換アミノ酸の記号をコンマ(,)で区切る。例えば仮想タンパク質が、位置100でアラニンに代わってグルタミン酸、グリシンまたはリシンで置換されている場合、表示はLuck−Ala100/Glu,Gly,LysまたはLuck−A100/E,G,Kとなる。
Claims (2)
- 配列番号2に記載のアミノ酸配列で示されるLucPplGRのアミノ酸配列が1か所または2か所で置換されている、該LucPplGRとは異なるアミノ酸配列を有する突然変異甲虫ルシフェラーゼであって、変異が、LucPplGR−R215G,R215T,R215M,R215P,V224L,V224H,V224G,I248N,H348A,H348C,H348N,H348Q,S247F/I248C,S247F/I248T,V224F/R215G,V224F/R215T,V224F/R215V,V224F/R215P,V224F/Q227E,V224F/L238V,V224F/L238M,V224F/S247G,V224F/S247H,V224F/S247Y,及びV224F/S247Fからなる群から選択される前記突然変異甲虫ルシフェラーゼ。
- 請求項1に記載の突然変異甲虫ルシフェラーゼをコードする配列部分からなるDNA分子。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US17708194A | 1994-01-03 | 1994-01-03 | |
US08/177,081 | 1994-01-03 |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP7518589A Division JPH08510387A (ja) | 1994-01-03 | 1995-01-03 | 突然変異体ルシフェラーゼ |
Publications (3)
Publication Number | Publication Date |
---|---|
JP2009039129A JP2009039129A (ja) | 2009-02-26 |
JP2009039129A5 JP2009039129A5 (ja) | 2009-04-09 |
JP5090304B2 true JP5090304B2 (ja) | 2012-12-05 |
Family
ID=22647117
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP7518589A Pending JPH08510387A (ja) | 1994-01-03 | 1995-01-03 | 突然変異体ルシフェラーゼ |
JP2008244507A Expired - Lifetime JP5090304B2 (ja) | 1994-01-03 | 2008-09-24 | 突然変異体ルシフェラーゼ |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP7518589A Pending JPH08510387A (ja) | 1994-01-03 | 1995-01-03 | 突然変異体ルシフェラーゼ |
Country Status (9)
Country | Link |
---|---|
US (2) | US6552179B1 (ja) |
EP (3) | EP0689587B1 (ja) |
JP (2) | JPH08510387A (ja) |
AT (1) | ATE391769T1 (ja) |
AU (1) | AU698424C (ja) |
CA (1) | CA2157476C (ja) |
DE (1) | DE69535744T2 (ja) |
ES (1) | ES2301160T3 (ja) |
WO (1) | WO1995018853A1 (ja) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US11306295B2 (en) | 2017-03-17 | 2022-04-19 | Toyo B-Net Co., Ltd. | Heat-resistant luciferase |
Families Citing this family (36)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE69633171T2 (de) * | 1995-01-20 | 2005-08-18 | The Secretary Of State For Defence, Salisbury | Mutierte luziferasen |
GB2311525B (en) * | 1995-01-20 | 1998-11-11 | Secr Defence | Mutant Luciferases |
GB9707486D0 (en) * | 1997-04-11 | 1997-05-28 | Secr Defence | Enzyme assays |
US6602677B1 (en) * | 1997-09-19 | 2003-08-05 | Promega Corporation | Thermostable luciferases and methods of production |
CA2324648C (en) * | 1998-03-27 | 2013-02-26 | Prolume, Ltd. | Luciferases, fluorescent proteins, nucleic acids encoding the luciferases and fluorescent proteins and the use thereof in diagnostics, high throughput screening and novelty items |
EP1925320A3 (en) | 1998-03-27 | 2008-09-03 | Prolume, Ltd. | Luciferases, fluorescent proteins, nucleic acids encoding the luciferases and fluorescent proteins and the use thereof in diagnostics |
GB9823468D0 (en) | 1998-10-28 | 1998-12-23 | Secr Defence | Novel enzyme |
JP3694181B2 (ja) * | 1999-01-05 | 2005-09-14 | キッコーマン株式会社 | 変異型ルシフェラーゼ、変異型ルシフェラーゼ遺伝子、新規な組み換え体dna及び変異型ルシフェラーゼの製造法 |
ES2292479T3 (es) * | 1999-10-26 | 2008-03-16 | The Secretary Of State For Defence | Luciferasa mutante. |
GB9925161D0 (en) * | 1999-10-26 | 1999-12-22 | Secr Defence | Novel enzyme |
US7879540B1 (en) | 2000-08-24 | 2011-02-01 | Promega Corporation | Synthetic nucleic acid molecule compositions and methods of preparation |
JP2006500115A (ja) | 2000-11-16 | 2006-01-05 | マイクロスフエリツクス・エル・エル・シー | 密封小線源治療用の可撓性および/または弾性シードまたはストランド |
GB0030727D0 (en) | 2000-12-15 | 2001-01-31 | Lumitech Uk Ltd | Methods and kits for detecting kinase activity |
GB2400659B8 (en) | 2003-04-17 | 2007-07-09 | Cambrex Bio Science Nottingham | Assay methods and materials |
EP2308978B8 (en) | 2003-10-10 | 2015-04-08 | Promega Corporation | Luciferase biosensor |
US7728118B2 (en) | 2004-09-17 | 2010-06-01 | Promega Corporation | Synthetic nucleic acid molecule compositions and methods of preparation |
AU2006281994B2 (en) * | 2005-08-19 | 2011-10-06 | Commonwealth Scientific And Industrial Research Organisation | Arachnocampa luciferases |
GB0517977D0 (en) * | 2005-09-02 | 2005-10-12 | Birkeland Innovasjon As | Transgenic animal model for identifying modulators of protein kinase |
US9359635B2 (en) | 2006-04-03 | 2016-06-07 | Promega Corporation | Permuted and nonpermuted luciferase biosensors |
WO2008109459A1 (en) * | 2007-03-02 | 2008-09-12 | University Of Massachusetts | Luciferins |
US20080299592A1 (en) * | 2007-03-02 | 2008-12-04 | Miller Stephen C | Red-Shifted Luciferase |
JP2008222084A (ja) * | 2007-03-14 | 2008-09-25 | Yamaha Motor Electronics Co Ltd | 電動ゴルフカーのブレーキ劣化検出方法及びこれを用いた電動ゴルフカー |
JP2008289475A (ja) * | 2007-04-27 | 2008-12-04 | National Institute Of Advanced Industrial & Technology | 最大発光波長がシフトした変異型ルシフェラーゼ |
EP2281046B1 (en) | 2008-05-19 | 2012-01-25 | Promega Corporation | LUCIFERASE BIOSENSORS FOR cAMP |
JP2010081908A (ja) * | 2008-10-02 | 2010-04-15 | Hitachi Ltd | 変異型のホタルルシフェラーゼ |
JP6038649B2 (ja) | 2009-05-01 | 2016-12-07 | プロメガ コーポレイションPromega Corporation | 増大した光出力を有する合成オプロフォルスルシフェラーゼ |
US9290794B2 (en) | 2010-05-11 | 2016-03-22 | Promega Corporation | Mutant protease biosensors with enhanced detection characteristics |
WO2011143339A1 (en) | 2010-05-11 | 2011-11-17 | Promega Corporation | Mutant protease biosensors with enhanced detection characteristics |
JP5889910B2 (ja) | 2010-11-02 | 2016-03-22 | プロメガ コーポレイションPromega Corporation | 新規セレンテラジン基質及び使用法 |
PT2635583E (pt) | 2010-11-02 | 2015-09-11 | Promega Corp | Derivados da coelenterazina e métodos de utilização da mesma |
US9927430B2 (en) | 2014-01-29 | 2018-03-27 | Promega Corporation | Pro-substrates for live cell applications |
JP6703484B2 (ja) | 2014-01-29 | 2020-06-03 | プロメガ コーポレイションPromega Corporation | 細胞による取り込み測定のための、標識用試薬としての、キノンでマスクされたプローブ |
JP6811173B2 (ja) * | 2014-09-11 | 2021-01-13 | プロメガ コーポレイションPromega Corporation | 増強された発光を産生するために赤外発光基質を活用するルシフェラーゼ配列 |
WO2016051517A1 (ja) | 2014-09-30 | 2016-04-07 | オリンパス株式会社 | オレンジ色の発光を示すルシフェラーゼ |
JP7175881B2 (ja) * | 2016-10-13 | 2022-11-21 | プロメガ コーポレイション | ルシフェリン分解産物による阻害に対して改善された耐性を持つ熱安定性ルシフェラーゼ |
WO2019236731A1 (en) * | 2018-06-05 | 2019-12-12 | Promega Corporation | Luciferase enzymes for use with thermostable luciferins in bioluminescent assays |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4968613A (en) | 1987-07-29 | 1990-11-06 | Kikkoman Corporation | Luciferase gene and novel recombinant DNA as well as a method of producing luciferase |
US5219737A (en) * | 1990-03-27 | 1993-06-15 | Kikkoman Corporation | Mutant luciferase of a firefly, mutant luciferase genes, recombinant dnas containing the genes and a method of producing mutant luciferase |
US5229285A (en) | 1991-06-27 | 1993-07-20 | Kikkoman Corporation | Thermostable luciferase of firefly, thermostable luciferase gene of firefly, novel recombinant dna, and process for the preparation of thermostable luciferase of firefly |
-
1995
- 1995-01-03 EP EP95910074A patent/EP0689587B1/en not_active Expired - Lifetime
- 1995-01-03 DE DE69535744T patent/DE69535744T2/de not_active Expired - Lifetime
- 1995-01-03 EP EP20080006460 patent/EP1978091A1/en not_active Ceased
- 1995-01-03 JP JP7518589A patent/JPH08510387A/ja active Pending
- 1995-01-03 CA CA002157476A patent/CA2157476C/en not_active Expired - Fee Related
- 1995-01-03 ES ES95910074T patent/ES2301160T3/es not_active Expired - Lifetime
- 1995-01-03 WO PCT/US1995/000108 patent/WO1995018853A1/en active Application Filing
- 1995-01-03 AT AT95910074T patent/ATE391769T1/de not_active IP Right Cessation
- 1995-01-03 AU AU18303/95A patent/AU698424C/en not_active Ceased
- 1995-01-03 EP EP08006578A patent/EP1978092A1/en not_active Withdrawn
- 1995-06-07 US US08/478,205 patent/US6552179B1/en not_active Expired - Lifetime
-
2002
- 2002-06-28 US US10/185,151 patent/US20030166905A1/en not_active Abandoned
-
2008
- 2008-09-24 JP JP2008244507A patent/JP5090304B2/ja not_active Expired - Lifetime
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US11306295B2 (en) | 2017-03-17 | 2022-04-19 | Toyo B-Net Co., Ltd. | Heat-resistant luciferase |
Also Published As
Publication number | Publication date |
---|---|
EP0689587B1 (en) | 2008-04-09 |
US6552179B1 (en) | 2003-04-22 |
ATE391769T1 (de) | 2008-04-15 |
JPH08510387A (ja) | 1996-11-05 |
EP0689587A4 (en) | 2001-08-08 |
ES2301160T3 (es) | 2008-06-16 |
AU1830395A (en) | 1995-08-01 |
AU698424B2 (en) | 1998-10-29 |
EP0689587A1 (en) | 1996-01-03 |
EP1978091A1 (en) | 2008-10-08 |
US20030166905A1 (en) | 2003-09-04 |
CA2157476A1 (en) | 1995-07-13 |
CA2157476C (en) | 2009-08-25 |
WO1995018853A1 (en) | 1995-07-13 |
AU698424C (en) | 2002-10-10 |
DE69535744T2 (de) | 2009-07-02 |
JP2009039129A (ja) | 2009-02-26 |
DE69535744D1 (de) | 2008-05-21 |
EP1978092A1 (en) | 2008-10-08 |
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