DE69133559T2 - Direkte Klonierung von PCR amplifizierten Nucleinsäuren - Google Patents
Direkte Klonierung von PCR amplifizierten Nucleinsäuren Download PDFInfo
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- DE69133559T2 DE69133559T2 DE69133559T DE69133559T DE69133559T2 DE 69133559 T2 DE69133559 T2 DE 69133559T2 DE 69133559 T DE69133559 T DE 69133559T DE 69133559 T DE69133559 T DE 69133559T DE 69133559 T2 DE69133559 T2 DE 69133559T2
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- cell
- nucleic acid
- dna
- pcr
- primer
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- 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
- C12N15/70—Vectors or expression systems specially adapted for E. coli
-
- 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
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
-
- 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
- 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
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6853—Nucleic acid amplification reactions using modified primers or templates
Landscapes
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- Chemical & Material Sciences (AREA)
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- Analytical Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Crystallography & Structural Chemistry (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
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| US589817 | 1984-03-15 | ||
| US58981790A | 1990-09-27 | 1990-09-27 |
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| DE69133559D1 DE69133559D1 (de) | 2007-03-22 |
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Family
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| DE69133559T Expired - Lifetime DE69133559T2 (de) | 1990-09-27 | 1991-09-27 | Direkte Klonierung von PCR amplifizierten Nucleinsäuren |
| DE69133389T Expired - Lifetime DE69133389T2 (de) | 1990-09-27 | 1991-09-27 | Direkte Klonierung von PCR amplifizierten Nukleinsäuren |
| DE69119083T Expired - Lifetime DE69119083T2 (de) | 1990-09-27 | 1991-09-27 | Direkte klonierung von pcr amplifizierten nukleinsäuren |
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| DE69133389T Expired - Lifetime DE69133389T2 (de) | 1990-09-27 | 1991-09-27 | Direkte Klonierung von PCR amplifizierten Nukleinsäuren |
| DE69119083T Expired - Lifetime DE69119083T2 (de) | 1990-09-27 | 1991-09-27 | Direkte klonierung von pcr amplifizierten nukleinsäuren |
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| AU (1) | AU8871891A (https=) |
| DE (3) | DE69133559T2 (https=) |
| WO (1) | WO1992006189A1 (https=) |
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| US5426039A (en) * | 1993-09-08 | 1995-06-20 | Bio-Rad Laboratories, Inc. | Direct molecular cloning of primer extended DNA containing an alkane diol |
| US5994058A (en) * | 1995-03-20 | 1999-11-30 | Genome International Corporation | Method for contiguous genome sequencing |
| WO1997048791A1 (en) * | 1996-06-18 | 1997-12-24 | Novagen, Inc. | Direct cloning of dna fragments |
| US5856144A (en) * | 1997-06-18 | 1999-01-05 | Novagen, Inc. | Direct cloning of DNA fragments |
| US6140086A (en) * | 1997-08-15 | 2000-10-31 | Fox; Donna K. | Methods and compositions for cloning nucleic acid molecules |
| EP1025217B1 (en) * | 1997-10-24 | 2006-10-04 | Invitrogen Corporation | Recombinational cloning using nucleic acids having recombination sites |
| US6358712B1 (en) | 1999-01-05 | 2002-03-19 | Trustee Of Boston University | Ordered gene assembly |
| CA2360011A1 (en) | 1999-01-05 | 2000-07-13 | Trustees Of Boston University | Ordered gene assembly |
| US20040005673A1 (en) | 2001-06-29 | 2004-01-08 | Kevin Jarrell | System for manipulating nucleic acids |
| JP2000253885A (ja) * | 1999-03-11 | 2000-09-19 | Jcr Pharmaceuticals Co Ltd | 任意プライマーｐｃｒを用いるｄｎａ断片クローニング方法 |
| US6280977B1 (en) * | 1999-03-24 | 2001-08-28 | Gene Therapy Systems | Method for generating transcriptionally active DNA fragments |
| US6709852B1 (en) * | 1999-06-22 | 2004-03-23 | Invitrogen Corporation | Rapid growing microorganisms for biotechnology applications |
| US6797509B1 (en) | 1999-07-09 | 2004-09-28 | Degussa-Huls Ag | Nucleotide sequences which code for the tal gene |
| US7205129B1 (en) * | 2000-02-28 | 2007-04-17 | Qiagen Gmbh | Method for reducing artifacts in nucleic acid amplification |
| KR100453279B1 (ko) * | 2000-04-27 | 2004-10-15 | 주식회사 서린바이오사이언스 | 양쪽 3'끝이 동일하거나 비상보적인 하나의 염기돌출부위를 갖는 벡터, ＥｃｌＨＫＩ 카세트 및 상기벡터와 ＥｃｌＨＫＩ 카세트의 이용방법 |
| GB0012233D0 (en) | 2000-05-19 | 2000-07-12 | Devgen Nv | Vector constructs |
| US7198924B2 (en) | 2000-12-11 | 2007-04-03 | Invitrogen Corporation | Methods and compositions for synthesis of nucleic acid molecules using multiple recognition sites |
| DE10109690A1 (de) | 2000-09-02 | 2002-03-14 | Degussa | Neue für das metY-Gen kodierende Nukleotidsequenzen |
| US6544782B1 (en) | 2000-11-13 | 2003-04-08 | Synthegen Systems | pREM: a positive selection vector system for direct PCR cloning |
| AU2715302A (en) | 2000-12-08 | 2002-06-18 | Invitrogen Corp | Methods and compositions for synthesis of nucleic acid molecules using multiple recognition sites |
| US7060434B2 (en) * | 2001-02-14 | 2006-06-13 | Council Of Scientific & Industrial Research | Probes for myctophid fish and a method for developing the same |
| US6566067B2 (en) * | 2001-02-14 | 2003-05-20 | Synthegen Systems, Inc. | High fidelity PCR cloning |
| DE10133928A1 (de) * | 2001-07-12 | 2003-01-23 | Bayer Cropscience Ag | Verfahren zum Herstellen von Deletionsmutanten |
| KR100441201B1 (ko) * | 2001-09-10 | 2004-07-23 | 대한민국 | 리포터 유전자를 포함하는 티벡터용 플라스미드 및 그 제조방법 |
| US6902914B2 (en) * | 2001-09-28 | 2005-06-07 | Sigma-Aldrich, Co. | Recombinant DNA processes using a dNTP mixture containing modified nucleotides |
| AU2002340234A1 (en) * | 2001-10-12 | 2003-04-22 | Modular Genetics, Inc. | System for manipulating nucleic acids |
| US20030180741A1 (en) | 2001-12-21 | 2003-09-25 | Holly Hogrefe | High fidelity DNA polymerase compositions and uses therefor |
| US20040241723A1 (en) * | 2002-03-18 | 2004-12-02 | Marquess Foley Leigh Shaw | Systems and methods for improving protein and milk production of dairy herds |
| WO2003091440A1 (en) * | 2002-04-25 | 2003-11-06 | Institute Of Zoology, Chinese Academy Of Sciences | Method for constructing a linear expression vector without cloning |
| US6803981B2 (en) * | 2002-05-24 | 2004-10-12 | Hannstar Display Corporation | Liquid crystal display having biased bending vertical alignment |
| US6586237B1 (en) * | 2002-05-24 | 2003-07-01 | Kyle David Yesland | Compositions and methods for cloning nucleic acids |
| 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 |
| CN101691577A (zh) | 2002-12-20 | 2010-04-07 | 梅坦诺米克斯有限公司 | 制备氨基酸的方法 |
| US20040214306A1 (en) * | 2003-01-23 | 2004-10-28 | Bloom Fredric R. | Rapid growing microorganisms for biotechnology applications |
| EP1606419A1 (en) | 2003-03-18 | 2005-12-21 | Quantum Genetics Ireland Limited | Systems and methods for improving protein and milk production of dairy herds |
| AU2004225520A1 (en) | 2003-03-25 | 2004-10-14 | Stratagene | DNA polymerase fusions and uses thereof |
| AU2004262656A1 (en) | 2003-08-01 | 2005-02-17 | Basf Plant Science Gmbh | Process for the production of fine chemicals in plants |
| CA2540348C (en) | 2003-09-29 | 2016-04-26 | Monsanto Technology Llc | Methods 0f enhancing stress tolerance in plants and compositions thereof |
| WO2005054438A2 (en) * | 2003-12-01 | 2005-06-16 | Invitrogen Corporation | Nucleic acid molecules containing recombination sites and methods of using the same |
| WO2005112544A2 (en) | 2004-02-19 | 2005-12-01 | The Governors Of The University Of Alberta | Leptin promoter polymorphisms and uses thereof |
| CN100465278C (zh) * | 2004-06-10 | 2009-03-04 | 清华大学 | 一种构建t载体的方法 |
| US20050281740A1 (en) * | 2004-06-16 | 2005-12-22 | Glen Gong | Imaging damaged lung tissue |
| EP2080769A3 (en) | 2004-07-02 | 2010-12-01 | Metanomics GmbH | Process for the production of fine chemicals |
| CA2585798A1 (en) | 2004-12-17 | 2006-06-17 | Metanomics Gmbh | Process for the control of production of fine chemicals |
| EP1836302B1 (en) * | 2004-12-23 | 2010-04-14 | Amersham Biosciences Corp. | Ligation-based rna amplification |
| BR122019024048B1 (pt) | 2004-12-28 | 2022-05-24 | Ajinomoto Co., Inc | Método para produzir ácido l-glutâmico |
| CA2594223C (en) * | 2005-01-03 | 2016-03-15 | The Government Of The United States Of America, As Represented By The Secretary Of The Department Of Health And Human Services, Centers For Disease Control And Prevention | Primer for nucleic acid detection |
| DE102005047596A1 (de) | 2005-10-05 | 2007-04-12 | Degussa Ag | Verfahren zur fermentativen Herstellung von L-Aminosäuren unter Verwendung coryneformer Bakterien |
| JP5088136B2 (ja) | 2005-10-18 | 2012-12-05 | 味の素株式会社 | コハク酸の製造方法 |
| CA2637600A1 (en) | 2006-01-17 | 2007-07-26 | Health Research, Inc. | Heteroduplex tracking assay |
| WO2007115059A2 (en) | 2006-03-29 | 2007-10-11 | Merial Limited | Vaccine against streptococci |
| CA2644273A1 (en) | 2006-04-05 | 2008-03-27 | Metanomics Gmbh | Process for the production of a fine chemical |
| US7723103B2 (en) * | 2007-01-08 | 2010-05-25 | Lucigen Corporation | Vectors, kits and methods for cloning DNA |
| DE102007005072A1 (de) | 2007-02-01 | 2008-08-07 | Evonik Degussa Gmbh | Verfahren zur fermentativen Herstellung von Cadaverin |
| US7881933B2 (en) * | 2007-03-23 | 2011-02-01 | Verizon Patent And Licensing Inc. | Age determination using speech |
| WO2008126896A1 (ja) | 2007-04-10 | 2008-10-23 | Ajinomoto Co., Inc. | 有機酸の製造方法 |
| US20110306099A1 (en) * | 2008-11-03 | 2011-12-15 | Matthew Beasley | Method of cloning dna |
| JP2012223091A (ja) | 2009-08-25 | 2012-11-15 | Ajinomoto Co Inc | Ｌ−アミノ酸の製造法 |
| US20120270271A1 (en) | 2009-12-16 | 2012-10-25 | University Of Washington | Toxin-Immunity System |
| US8557972B2 (en) * | 2009-12-21 | 2013-10-15 | University Of Washington Through Its Center For Commercialization | Compositions and methods for transfection of RNA and controlled stabilization of transfected RNA |
| WO2012113900A1 (en) | 2011-02-24 | 2012-08-30 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Igfbp-3 derivatives and uses thereof |
| RU2507266C2 (ru) * | 2011-11-16 | 2014-02-20 | Общество с ограниченной ответственностью "Лаборатория медицинской биотехнологии" (ООО "ЛМБТ") | НЕТРАНСКРИБИРУЕМЫЙ ПЛАЗМИДНЫЙ ВЕКТОР pBL-T ДЛЯ КЛОНИРОВАНИЯ СЛОЖНЫХ ГЕНОМНЫХ ПОСЛЕДОВАТЕЛЬНОСТЕЙ И МНОГОМОДУЛЬНОЙ СБОРКИ ГЕННО-ИНЖЕНЕРНЫХ КОНСТРУКЦИЙ (ВАРИАНТЫ), И НАБОР, СОДЕРЖАЩИЙ УКАЗАННЫЙ ВЕКТОР |
| US10358655B1 (en) | 2012-06-27 | 2019-07-23 | University Of Utah Research Foundation | Attenuated protein expression vectors and associated methods |
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| CN115920796A (zh) | 2015-12-01 | 2023-04-07 | 特韦斯特生物科学公司 | 功能化表面及其制备 |
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| CA3100739A1 (en) | 2018-05-18 | 2019-11-21 | Twist Bioscience Corporation | Polynucleotides, reagents, and methods for nucleic acid hybridization |
| WO2020139871A1 (en) | 2018-12-26 | 2020-07-02 | Twist Bioscience Corporation | Highly accurate de novo polynucleotide synthesis |
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| JP2022522668A (ja) | 2019-02-26 | 2022-04-20 | ツイスト バイオサイエンス コーポレーション | 抗体を最適化するための変異体核酸ライブラリ |
| US11332738B2 (en) | 2019-06-21 | 2022-05-17 | Twist Bioscience Corporation | Barcode-based nucleic acid sequence assembly |
| EP4034566A4 (en) | 2019-09-23 | 2024-01-24 | Twist Bioscience Corporation | VARIANT NUCLEIC ACID BANKS FOR CRTH2 |
| JP2022548783A (ja) | 2019-09-23 | 2022-11-21 | ツイスト バイオサイエンス コーポレーション | 単一ドメイン抗体のバリアント核酸ライブラリー |
| BR112022011235A2 (pt) | 2019-12-09 | 2022-12-13 | Twist Bioscience Corp | Bibliotecas de variantes de ácido nucleico para receptores de adenosina |
| WO2022167396A1 (en) | 2021-02-02 | 2022-08-11 | Biocopy Gmbh | Method for identifying emerging pathogen mutations enabling host cell entry and immune evasion |
Family Cites Families (3)
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|---|---|---|---|---|
| US4965188A (en) * | 1986-08-22 | 1990-10-23 | Cetus Corporation | Process for amplifying, detecting, and/or cloning nucleic acid sequences using a thermostable enzyme |
| WO1997048791A1 (en) * | 1996-06-18 | 1997-12-24 | Novagen, Inc. | Direct cloning of dna fragments |
| US5856144A (en) * | 1997-06-18 | 1999-01-05 | Novagen, Inc. | Direct cloning of DNA fragments |
-
1991
- 1991-09-27 DE DE69133559T patent/DE69133559T2/de not_active Expired - Lifetime
- 1991-09-27 DE DE69133389T patent/DE69133389T2/de not_active Expired - Lifetime
- 1991-09-27 DE DE69119083T patent/DE69119083T2/de not_active Expired - Lifetime
- 1991-09-27 EP EP91919634A patent/EP0550693B1/en not_active Expired - Lifetime
- 1991-09-27 EP EP03023664A patent/EP1396540B1/en not_active Expired - Lifetime
- 1991-09-27 AU AU88718/91A patent/AU8871891A/en not_active Abandoned
- 1991-09-27 WO PCT/US1991/007147 patent/WO1992006189A1/en not_active Ceased
- 1991-09-27 EP EP96250006A patent/EP0738779B1/en not_active Expired - Lifetime
- 1991-09-27 EP EP07002538A patent/EP1790723A3/en not_active Withdrawn
-
1993
- 1993-09-09 US US08/119,313 patent/US5487993A/en not_active Expired - Lifetime
-
1996
- 1996-07-18 US US08/683,237 patent/US5827657A/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| EP0738779A2 (en) | 1996-10-23 |
| DE69133389T2 (de) | 2005-06-02 |
| DE69119083T2 (de) | 1996-10-17 |
| DE69119083D1 (de) | 1996-05-30 |
| HK1064124A1 (en) | 2005-01-21 |
| EP1790723A3 (en) | 2008-10-22 |
| EP0550693A1 (en) | 1993-07-14 |
| US5487993A (en) | 1996-01-30 |
| EP0738779A3 (en) | 1997-05-07 |
| EP0550693B1 (en) | 1996-04-24 |
| US5827657A (en) | 1998-10-27 |
| EP1396540B1 (en) | 2007-02-07 |
| EP1396540A2 (en) | 2004-03-10 |
| EP1790723A2 (en) | 2007-05-30 |
| DE69133389D1 (de) | 2004-06-24 |
| EP0738779B1 (en) | 2004-05-19 |
| EP1396540A3 (en) | 2004-06-09 |
| WO1992006189A1 (en) | 1992-04-16 |
| DE69133559D1 (de) | 2007-03-22 |
| AU8871891A (en) | 1992-04-28 |
| EP0550693A4 (https=) | 1994-04-27 |
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