JP2000501616A - テルモアナエロバクター・テルモヒドロスルフリカスからの熱安定性dnaポリメラーゼおよびエキソヌクレアーゼ活性が除去されているその変異体酵素 - Google Patents
テルモアナエロバクター・テルモヒドロスルフリカスからの熱安定性dnaポリメラーゼおよびエキソヌクレアーゼ活性が除去されているその変異体酵素Info
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- JP2000501616A JP2000501616A JP9522273A JP52227397A JP2000501616A JP 2000501616 A JP2000501616 A JP 2000501616A JP 9522273 A JP9522273 A JP 9522273A JP 52227397 A JP52227397 A JP 52227397A JP 2000501616 A JP2000501616 A JP 2000501616A
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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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
- C12N9/1241—Nucleotidyltransferases (2.7.7)
- C12N9/1252—DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase
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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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
- C12N9/1241—Nucleotidyltransferases (2.7.7)
- C12N9/1276—RNA-directed DNA polymerase (2.7.7.49), i.e. reverse transcriptase or telomerase
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6869—Methods for sequencing
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- Bioinformatics & Cheminformatics (AREA)
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- Biotechnology (AREA)
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- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
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- Biomedical Technology (AREA)
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- Analytical Chemistry (AREA)
- Biophysics (AREA)
- Immunology (AREA)
- Enzymes And Modification Thereof (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Saccharide Compounds (AREA)
Abstract
Description
Claims (1)
- 【特許請求の範囲】 1.アミノ酸配列において、テルモアナエロバクター・テルモヒドロスルフリカ スのDNAポリメラーゼの少なくとも40個の連続するアミノ酸配列と少なくと も80%のホモロジーを有する、酵素的に活性なDNAポリメラーゼまたはその フラグメント。 2.前記DNAポリメラーゼが、エキソヌクレアーゼ活性が除去されているもの である、請求項1記載のポリメラーゼ。 3.前記ポリメラーゼが、アミノ末端から1個またはそれ以上のアミノ酸が欠失 されているかまたはアミノ酸が変更されて、前記エキソヌクレアーゼ活性が除去 されている、請求項2記載のポリメラーゼ。 4.前記ポリメラーゼが、706位においてフェニルアラニンがチロシンに置き 換えられたものである、請求項1記載のポリメラーゼ。 5.前記ポリメラーゼが、アミノ末端から1個またはそれ以上のアミノ酸が欠失 され、706位においてフェニルアラニンがチロシンに置き換えられたものであ る、請求項1記載のポリメラーゼ。 6.前記ポリメラーゼが、1個またはそれ以上のアミノ酸が変更されてエキソヌ クレアーゼ活性が除去されており、706位においてフェニルアラニンがチロシ ンに置き換えられたものである、請求項1記載のポリメラーゼ。 7.請求項1−6のいずれかに記載の精製されたDNAポリメラーゼを緩衝液中 に含む、安定な酵素的組成物。 8.請求項1−6のいずれかに記載のDNAポリメラーゼをコードする精製され た核酸。 9.請求項1−6のいずれかに記載のDNAポリメラーゼをコードする核酸配列 を含有するベクターを含む宿主細胞。 10.鎖置換増幅において用いるための、請求項1−6のいずれかに記載のDN Aポリメラーゼおよび熱安定性制限酵素。 11.アミノ酸配列においてテルモアナエロバクター・テルモヒドロスルフリカ スのDNAポリメラーゼの少なくとも40個の連続するアミノ酸配列に少なくと も80%ホモロジーを有するDNAポリメラーゼまたはそのフラグメントを用い て、鎖置換により核酸フラグメントを生成し増幅する方法。 12.前記DNAポリメラーゼが、エキソヌクレアーゼ活性が除去されたもので ある、請求項11記載の方法。 13.請求項1−6のいずれかに記載のDNAポリメラーゼおよび熱安定性制限 酵素を含む、DNAの鎖置換増幅のためのキット。 14.DNAをシークエンシングし、このことにより、DNAテンプレートのヌ クレオチド塩基配列の少なくとも一部を決定することができる方法であって、請 求項4、5、または6のいずれかに記載のDNAポリメラーゼを少なくとも1つ の鎖停止剤の存在下で用いて、前記DNAテンプレートから鎖停止されたフラグ メントを生成し、そして前記フラグメントのサイズから前記DNAテンプレート の少なくとも一部の配列を決定する、 の各工程を含む方法。 15.請求項4、5、または6のいずれかに記載のDNAポリメラーゼおよびピ ロホスファターゼを含む、DNAシークエンシング用のキット。 16.オリゴヌクレオチドプライマー、請求項1−6のいずれかに記載のポリメ ラーゼ、および1種類から4種類のデオキシリボヌクレオシドリン酸を提供する ことによりcDNAを製造する方法。 17.テルモアナエロバクター・テルモヒドロスルフリカスからの精製されたリ バーストランスクリプターゼ。 18.リバーストランスクリプターゼとして有用な、請求項1記載のDNAポリ メラーゼ。 19.リバーストランスクリプターゼとして有用な、請求項2記載のDNAポリ メラーゼ。 20.請求項1記載のDNAポリメラーゼおよび単一の反応容器中でポリメラー ゼ連鎖反応を実施するのに適したポリメラーゼを用いる逆転写/ポリメラーゼ連 鎖反応の方法。 21.前記DNAポリメラーゼが、エキソヌクレアーゼ活性が除去されたもので ある、請求項20記載の方法。 22.ポリメラーゼ連鎖反応を実施するのに適した前記ポリメラーゼがTaqD NAポリメラーゼである、請求項20記載の方法。 23.請求項1記載のDNAポリメラーゼおよびポリメラーゼ連鎖反応を実施す るのに適したポリメラーゼを含む、逆転写/ポリメラーゼ連鎖反応用のキット。 24.前記DNAポリメラーゼが、エキソヌクレアーゼ活性が除去されたもので ある、請求項23記載のキット。 25.ポリメラーゼ連鎖反応を実施するのに適した前記ポリメラーゼがTaqD NAポリメラーゼである、請求項23記載のキット。 26.請求項1記載のDNAポリメラーゼおよびポリメラーゼ連鎖反応を実施す るのに適したポリメラーゼを含む、RT/PCRにおいて用いるための溶液。 27.前記DNAポリメラーゼが、エキソヌクレアーゼ活性が除去されたもので ある、請求項26記載の溶液。 28.ポリメラーゼ連鎖反応を実施するのに適した前記ポリメラーゼがTaqD NAポリメラーゼである、請求項26記載の溶液。 29.請求項1−6のいずれかに記載のDNAポリメラーゼおよび熱安定性制限 酵素を含む、DNAの鎖置換増幅のための溶液。 30.請求項4、5、または6のいずれかに記載のDNAポリメラーゼおよびピ ロホスファターゼを含む、DNAのシークエンシングのための溶液。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US868895P | 1995-12-15 | 1995-12-15 | |
US60/008,688 | 1995-12-15 | ||
PCT/US1996/020225 WO1997021821A1 (en) | 1995-12-15 | 1996-12-13 | Thermostable dna polymerase from thermoanaerobacter thermohydrosulfuricus and mutant enzymes thereof with exonuclease activity removed |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2000501616A true JP2000501616A (ja) | 2000-02-15 |
JP4073038B2 JP4073038B2 (ja) | 2008-04-09 |
Family
ID=21733102
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP52227397A Expired - Fee Related JP4073038B2 (ja) | 1995-12-15 | 1996-12-13 | テルモアナエロバクター・テルモヒドロスルフリカスからの熱安定性dnaポリメラーゼおよびエキソヌクレアーゼ活性が除去されているその変異体酵素 |
Country Status (8)
Country | Link |
---|---|
US (1) | US5744312A (ja) |
EP (1) | EP0866868B1 (ja) |
JP (1) | JP4073038B2 (ja) |
AT (1) | ATE203056T1 (ja) |
CA (1) | CA2240334C (ja) |
DE (1) | DE69613856T2 (ja) |
ES (1) | ES2162132T3 (ja) |
WO (1) | WO1997021821A1 (ja) |
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US7875440B2 (en) * | 1998-05-01 | 2011-01-25 | Arizona Board Of Regents | Method of determining the nucleotide sequence of oligonucleotides and DNA molecules |
US6780591B2 (en) | 1998-05-01 | 2004-08-24 | Arizona Board Of Regents | Method of determining the nucleotide sequence of oligonucleotides and DNA molecules |
ATE371022T1 (de) | 1998-06-17 | 2007-09-15 | Ge Healthcare Bio Sciences | Fy7 polymerase |
US6406891B1 (en) | 1998-09-28 | 2002-06-18 | Board Of Regents, The University Of Texas System | Dual RT procedure for cDNA synthesis |
US6238868B1 (en) * | 1999-04-12 | 2001-05-29 | Nanogen/Becton Dickinson Partnership | Multiplex amplification and separation of nucleic acid sequences using ligation-dependant strand displacement amplification and bioelectronic chip technology |
US6818395B1 (en) | 1999-06-28 | 2004-11-16 | California Institute Of Technology | Methods and apparatus for analyzing polynucleotide sequences |
US6692918B2 (en) | 1999-09-13 | 2004-02-17 | Nugen Technologies, Inc. | Methods and compositions for linear isothermal amplification of polynucleotide sequences |
AU783873B2 (en) | 1999-09-13 | 2005-12-15 | Nugen Technologies, Inc. | Methods and compositions for linear isothermal amplification of polynucleotide sequences |
MXPA02012735A (es) | 2000-06-26 | 2004-04-20 | Nugen Tgechnologies Inc | Metodos y composiciones para la amplificacion de acido nucleico a base de transcripcion. |
US7846733B2 (en) * | 2000-06-26 | 2010-12-07 | Nugen Technologies, Inc. | Methods and compositions for transcription-based nucleic acid amplification |
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US7397546B2 (en) * | 2006-03-08 | 2008-07-08 | Helicos Biosciences Corporation | Systems and methods for reducing detected intensity non-uniformity in a laser beam |
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US4683195A (en) * | 1986-01-30 | 1987-07-28 | Cetus Corporation | Process for amplifying, detecting, and/or-cloning nucleic acid sequences |
US5405774A (en) * | 1986-08-22 | 1995-04-11 | Hoffmann-La Roche Inc. | DNA encoding a mutated thermostable nucleic acid polymerase enzyme from thermus species sps17 |
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-
1996
- 1996-12-13 EP EP96944875A patent/EP0866868B1/en not_active Expired - Lifetime
- 1996-12-13 JP JP52227397A patent/JP4073038B2/ja not_active Expired - Fee Related
- 1996-12-13 AT AT96944875T patent/ATE203056T1/de not_active IP Right Cessation
- 1996-12-13 CA CA002240334A patent/CA2240334C/en not_active Expired - Fee Related
- 1996-12-13 WO PCT/US1996/020225 patent/WO1997021821A1/en active IP Right Grant
- 1996-12-13 DE DE69613856T patent/DE69613856T2/de not_active Expired - Lifetime
- 1996-12-13 US US08/766,014 patent/US5744312A/en not_active Expired - Lifetime
- 1996-12-13 ES ES96944875T patent/ES2162132T3/es not_active Expired - Lifetime
Also Published As
Publication number | Publication date |
---|---|
EP0866868B1 (en) | 2001-07-11 |
ATE203056T1 (de) | 2001-07-15 |
JP4073038B2 (ja) | 2008-04-09 |
DE69613856D1 (de) | 2001-08-16 |
ES2162132T3 (es) | 2001-12-16 |
CA2240334A1 (en) | 1997-06-19 |
DE69613856T2 (de) | 2002-04-04 |
US5744312A (en) | 1998-04-28 |
CA2240334C (en) | 2008-07-22 |
WO1997021821A1 (en) | 1997-06-19 |
EP0866868A1 (en) | 1998-09-30 |
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