JP5638491B2 - Dnaクローニングベクタープラスミドのダイナミックなベクター構築のための方法 - Google Patents
Dnaクローニングベクタープラスミドのダイナミックなベクター構築のための方法 Download PDFInfo
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- JP5638491B2 JP5638491B2 JP2011204896A JP2011204896A JP5638491B2 JP 5638491 B2 JP5638491 B2 JP 5638491B2 JP 2011204896 A JP2011204896 A JP 2011204896A JP 2011204896 A JP2011204896 A JP 2011204896A JP 5638491 B2 JP5638491 B2 JP 5638491B2
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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
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/64—General methods for preparing the vector, for introducing it into the cell or for selecting the vector-containing host
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
- C07H21/04—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with deoxyribosyl as saccharide radical
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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
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/66—General methods for inserting a gene into a vector to form a recombinant vector using cleavage and ligation; Use of non-functional linkers or adaptors, e.g. linkers containing the sequence for a restriction endonuclease
-
- 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
- C12N15/63—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
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
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- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Cell Biology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Description
5’……G−A−A−T−T−C……3’
3’……C−T−T−A−A−G……5’
5’……G/A−A−T−T−C……3’
3’……C−T−T−A−A/G……5’
その結果二本鎖DNA分子が切断されて、結果として生じるヌクレオチドの配置を新しく形成される「端部」で有することとなる:
5’……G3’ 5’A−A−T−T−C……3’
3’……C−T−T−A−A5’ 3’G……5’。
5’...TAGGGATAA/CAGGGTAAT...3’
3’...ATCCC/TATTGTCCCATTA...5’。
5’...TAGGGATAA CCCTA...3’
3’...ATCCCAATAGGGAT...5’
5’−CCAGATAA
3’−GGTC
CAGGGTAAT//ATTACCCTGTTAT
TATTGTCCCATTA//TAATGGGAC
GTGG−3’
AATACACC−5’
AseI、BamHI、BglII、BlpI、BstXI、EcoRI、HincII、HindIII、NcoI、PstI、SacI、SacII、SphI、StuI、XbaI
AarI、AatII、AflII、AgeI、ApaLI、AvrII、BseAI、BspDI、BspEI、BstBI、ClaI、EagI、EcoO109I、EcoRV、HpaI、KpnI、MfeI、NarI、NdeI、NgoMIV、NheI、NsiI、PmlI、SexAI、SmaI、SpeI、XhoI
AclI、AscI、AsiSI、BsiWI、FseI、MluI、NotI、NruI、PacI、PmeI、PvuI、RsrII、SalI、SbfI、SfiI、SgrAI、SnaBI、SwaI、PI−SceI、I−SceI、I−CeuI、PI−PspI、I−PpoI、I−TliI
pCMV−EGFP−SV40
pCMV−EGFP−hGH
pCMV−ルシフェラーゼ−SV40
pCMV−ルシフェラーゼ−hGH
pSPC−EGFP−SV40
pSPC−EGFP−hGH
pSPC−ルシフェラーゼ−SV40
pSPC−ルシフェラーゼ−hGH
Claims (2)
- トランスジーンのアレイを同時に合成するための方法であって、
a)第一、及び第二のドッキングポイントを有する一次クローニングベクタープラスミドを用意する工程であって、前記第一、及び第二のドッキングポイントの各々が、少なくとも一つの6ヌクレオチドよりも大きな制限エンドヌクレアーゼ部位を含み、前記クローニングベクタープラスミドが、前記第一のドッキングポイントの5’末端の上流に位置する順方向のユニークなホーミングエンドヌクレアーゼ部位及び前記第二のドッキングポイントの3’末端の下流に位置する逆方向のユニークなホーミングエンドヌクレアーゼ部位を更に含む工程;
b)少なくとも一のプロモーターヌクレオチド配列をプロモーターシャトルベクターに導入する工程;
c)少なくとも一の発現ヌクレオチド配列を発現シャトルベクターに導入する工程;
d)少なくとも一の調節ヌクレオチド配列を調節シャトルベクターに導入する工程;
e)前記プロモーター、発現、及び調節ヌクレオチド配列を、同時に、前記プロモーター、発現、及び調節シャトルベクターから、前記クローニングベクタープラスミドの前記第一及び第二のドッキングポイントの間に結合させる工程;
f)前記ホーミングエンドヌクレアーゼ部位を含む二次クローニングベクタープラスミドを用意する工程;及び、
g)前記プロモーターヌクレオチド配列、前記発現ヌクレオチド配列及び前記調節ヌクレオチド配列を、前記二次クローニングベクタープラスミドの前記ホーミングエンドヌクレアーゼ部位に結合させる工程を含む方法。 - 前記プロモーターヌクレオチド配列、前記発現ヌクレオチド配列及び前記調節ヌクレオチド配列がde novo合成、組換え、及びPCRターミネーター突出クローニングからなる群から選ばれる方法により作成される請求項1に記載の方法。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US57201104P | 2004-05-18 | 2004-05-18 | |
US60/572,011 | 2004-05-18 |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
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JP2007527380A Division JP2007537755A (ja) | 2004-05-18 | 2005-05-18 | Dnaクローニングベクタープラスミドのダイナミックなベクター構築のための方法 |
Publications (3)
Publication Number | Publication Date |
---|---|
JP2012024099A JP2012024099A (ja) | 2012-02-09 |
JP2012024099A5 JP2012024099A5 (ja) | 2013-01-31 |
JP5638491B2 true JP5638491B2 (ja) | 2014-12-10 |
Family
ID=35450900
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
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JP2007527380A Pending JP2007537755A (ja) | 2004-05-18 | 2005-05-18 | Dnaクローニングベクタープラスミドのダイナミックなベクター構築のための方法 |
JP2011204896A Expired - Fee Related JP5638491B2 (ja) | 2004-05-18 | 2011-09-20 | Dnaクローニングベクタープラスミドのダイナミックなベクター構築のための方法 |
Family Applications Before (1)
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JP2007527380A Pending JP2007537755A (ja) | 2004-05-18 | 2005-05-18 | Dnaクローニングベクタープラスミドのダイナミックなベクター構築のための方法 |
Country Status (9)
Country | Link |
---|---|
US (4) | US20090170727A1 (ja) |
EP (2) | EP1751298A4 (ja) |
JP (2) | JP2007537755A (ja) |
CN (1) | CN1981047B (ja) |
AU (1) | AU2005248371B2 (ja) |
CA (1) | CA2567337C (ja) |
ES (1) | ES2603062T3 (ja) |
NZ (2) | NZ583974A (ja) |
WO (1) | WO2005116231A1 (ja) |
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US20080050808A1 (en) * | 2002-10-09 | 2008-02-28 | Reed Thomas D | DNA modular cloning vector plasmids and methods for their use |
US7785871B2 (en) * | 2002-10-09 | 2010-08-31 | Intrexon Corporation | DNA cloning vector plasmids and methods for their use |
CN1981047B (zh) | 2004-05-18 | 2013-08-21 | 英特拉克森公司 | Dna克隆载体质粒的动态载体装配方法 |
US8153598B2 (en) | 2005-10-19 | 2012-04-10 | Intrexon Corporation | PKD ligands and polynucleotides encoding PKD ligands |
US7943732B2 (en) | 2006-06-05 | 2011-05-17 | Intrexon Corporation | AKT ligands and polynucleotides encoding AKT ligands |
JP2009544711A (ja) | 2006-07-26 | 2009-12-17 | イントレクソン コーポレイション | 疾病の治療方法及び疾病治療用組成物 |
US8993263B2 (en) | 2006-08-07 | 2015-03-31 | Intrexon Corporation | PKA ligands and polynucleotides encoding PKA ligands |
US8999666B2 (en) | 2006-08-09 | 2015-04-07 | Intrexon Corporation | PKC ligands and polynucleotides encoding PKC ligands |
US7897394B2 (en) | 2006-09-21 | 2011-03-01 | Intrexon Corporation | Endoplasmic reticulum localization signals |
US8729225B2 (en) | 2006-11-13 | 2014-05-20 | Intrexon Corporation | Glycogen synthase kinase heteropolyligand polypeptide |
US8283445B2 (en) | 2006-11-13 | 2012-10-09 | Intrexon Corporation | Extracellular-signal-regulated-kinase (ERK) heteropolyligand polypeptide |
US20090181458A1 (en) | 2006-12-04 | 2009-07-16 | Thomas David Reed | Tubulo-vesicular structure localization signals |
US7705122B2 (en) | 2006-12-04 | 2010-04-27 | Intrexon Corporation | mTOR ligands and polynucleotides encoding mTOR ligands |
DK2126064T3 (da) | 2007-03-27 | 2012-09-10 | Intrexon Corp | MEK-ligander og polynukleotider til kodning af MEK-ligander |
WO2008137681A2 (en) | 2007-05-02 | 2008-11-13 | Intrexon Corporation | Pp1 ligands |
EP2185730A4 (en) | 2007-08-23 | 2010-10-27 | Intrexon Corp | METHOD AND COMPOSITIONS FOR SICKNESS DIAGNOSIS |
JP5514727B2 (ja) | 2007-09-26 | 2014-06-04 | イントレキソン コーポレーション | 合成5’utr、発現ベクター、および導入遺伝子の発現を増加させる方法 |
JP2010540534A (ja) | 2007-09-28 | 2010-12-24 | イントレキソン コーポレーション | 生体治療分子の発現のための治療遺伝子スイッチ構築物およびバイオリアクター、ならびにその使用 |
KR101655487B1 (ko) | 2007-10-08 | 2016-09-08 | 인트렉손 코포레이션 | 가공된 수지상 세포 및 암의 치료를 위한 이의 용도 |
AU2009204464B2 (en) | 2008-01-09 | 2014-12-04 | Intrexon Corporation | Therapeutic inhibitors of PAI-1 function and methods of their use |
TWI688395B (zh) | 2010-03-23 | 2020-03-21 | 英翠克頌公司 | 條件性表現治療性蛋白質之載體、包含該載體之宿主細胞及彼等之用途 |
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FR2986536A1 (fr) * | 2012-02-08 | 2013-08-09 | Lfb Biotechnologies | Unites de transcription et leur utilisation dans des vecteurs d'expression (cho) |
WO2013191950A2 (en) | 2012-06-22 | 2013-12-27 | Monsanto Technology Llc | Unique modular vector design |
US10253358B2 (en) | 2013-11-04 | 2019-04-09 | Exact Sciences Development Company, Llc | Multiple-control calibrators for DNA quantitation |
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2005
- 2005-05-18 CN CN2005800149458A patent/CN1981047B/zh not_active Expired - Fee Related
- 2005-05-18 EP EP20050751019 patent/EP1751298A4/en not_active Withdrawn
- 2005-05-18 NZ NZ583974A patent/NZ583974A/en not_active IP Right Cessation
- 2005-05-18 CA CA2567337A patent/CA2567337C/en not_active Expired - Fee Related
- 2005-05-18 NZ NZ551447A patent/NZ551447A/en not_active IP Right Cessation
- 2005-05-18 US US11/569,335 patent/US20090170727A1/en not_active Abandoned
- 2005-05-18 AU AU2005248371A patent/AU2005248371B2/en not_active Ceased
- 2005-05-18 ES ES11009458.8T patent/ES2603062T3/es active Active
- 2005-05-18 JP JP2007527380A patent/JP2007537755A/ja active Pending
- 2005-05-18 WO PCT/US2005/017272 patent/WO2005116231A1/en active Application Filing
- 2005-05-18 EP EP11009458.8A patent/EP2484772B1/en not_active Not-in-force
-
2007
- 2007-08-20 US US11/841,380 patent/US20090226976A1/en not_active Abandoned
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2011
- 2011-09-20 JP JP2011204896A patent/JP5638491B2/ja not_active Expired - Fee Related
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2014
- 2014-11-04 US US14/532,088 patent/US9115361B2/en not_active Expired - Fee Related
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2015
- 2015-08-13 US US14/825,195 patent/US10036026B2/en not_active Expired - Fee Related
Also Published As
Publication number | Publication date |
---|---|
EP1751298A4 (en) | 2009-11-11 |
CN1981047B (zh) | 2013-08-21 |
ES2603062T3 (es) | 2017-02-23 |
NZ551447A (en) | 2010-08-27 |
US9115361B2 (en) | 2015-08-25 |
AU2005248371A1 (en) | 2005-12-08 |
CN1981047A (zh) | 2007-06-13 |
NZ583974A (en) | 2011-11-25 |
US10036026B2 (en) | 2018-07-31 |
AU2005248371B2 (en) | 2011-12-15 |
US20180023089A1 (en) | 2018-01-25 |
US20150057192A1 (en) | 2015-02-26 |
US20090170727A1 (en) | 2009-07-02 |
EP2484772A1 (en) | 2012-08-08 |
EP2484772B1 (en) | 2016-08-17 |
EP1751298A1 (en) | 2007-02-14 |
CA2567337A1 (en) | 2005-12-08 |
JP2007537755A (ja) | 2007-12-27 |
JP2012024099A (ja) | 2012-02-09 |
WO2005116231A1 (en) | 2005-12-08 |
CA2567337C (en) | 2017-12-12 |
US20090226976A1 (en) | 2009-09-10 |
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