WO2009155464A3 - Mutated and chemically modified thermally stable dna polymerases - Google Patents

Mutated and chemically modified thermally stable dna polymerases Download PDF

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Publication number
WO2009155464A3
WO2009155464A3 PCT/US2009/047862 US2009047862W WO2009155464A3 WO 2009155464 A3 WO2009155464 A3 WO 2009155464A3 US 2009047862 W US2009047862 W US 2009047862W WO 2009155464 A3 WO2009155464 A3 WO 2009155464A3
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WO
WIPO (PCT)
Prior art keywords
polymerase
chemically modified
thermally stable
mutated
stable dna
Prior art date
Application number
PCT/US2009/047862
Other languages
French (fr)
Other versions
WO2009155464A2 (en
Inventor
Lei Xi
Original Assignee
Life Technologies Corporation
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Life Technologies Corporation filed Critical Life Technologies Corporation
Priority to EP09767773A priority Critical patent/EP2300613A4/en
Publication of WO2009155464A2 publication Critical patent/WO2009155464A2/en
Publication of WO2009155464A3 publication Critical patent/WO2009155464A3/en

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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
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/6848—Nucleic acid amplification reactions characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/26—Preparation of nitrogen-containing carbohydrates
    • C12P19/28—N-glycosides
    • C12P19/30—Nucleotides
    • C12P19/34—Polynucleotides, e.g. nucleic acids, oligoribonucleotides

Abstract

A mutant thermally stable DNA polymerase having at least one Lysine to Arginine substitution corresponding to at least one of amino acid position 505, 540 or 542 of Thermus aquaticus Taq polymerase A. Said polymerase can also be chemically modified to substantially reduced polymerase activity at ambient temperatures, wherein the chemical modification modifies at least one Lysine residue of the polymerase and said reduced polymerase activity is reversible upon heating at a temperature of at least 50 °C. Methods for using and kits containing the mutant DNA polymerase and chemically modified, mutant DNA polymerase for primer extension and amplification, respectively, are also envisioned.
PCT/US2009/047862 2008-06-18 2009-06-18 Mutated and chemically modified thermally stable dna polymerases WO2009155464A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP09767773A EP2300613A4 (en) 2008-06-18 2009-06-18 Mutated and chemically modified thermally stable dna polymerases

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US7375408P 2008-06-18 2008-06-18
US61/073,754 2008-06-18

Publications (2)

Publication Number Publication Date
WO2009155464A2 WO2009155464A2 (en) 2009-12-23
WO2009155464A3 true WO2009155464A3 (en) 2010-05-06

Family

ID=41434700

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2009/047862 WO2009155464A2 (en) 2008-06-18 2009-06-18 Mutated and chemically modified thermally stable dna polymerases

Country Status (2)

Country Link
EP (1) EP2300613A4 (en)
WO (1) WO2009155464A2 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP7470216B2 (en) 2020-05-29 2024-04-17 ジェノテック コーポレイション DNA polymerase mutants with improved tolerance to genetic mutations

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102245761A (en) 2008-11-03 2011-11-16 卡帕生物系统 Modified type A DNA polymerases
WO2012097318A2 (en) 2011-01-14 2012-07-19 Kap Abiosystems Modified dna polymerases for improved amplification
US9447388B2 (en) * 2011-07-12 2016-09-20 Kyushu University, National University Corporation DNA polymerases
WO2013160316A1 (en) * 2012-04-23 2013-10-31 Dsm Ip Assets B.V. Polypeptide expression method
DE102012216193B4 (en) 2012-09-12 2020-07-30 Continental Automotive Gmbh Method and device for operating a motor vehicle component using gestures
EP3757119A1 (en) * 2015-11-16 2020-12-30 Ubiprotein, Corp. A method for extending half-life of a protein
WO2018203582A1 (en) * 2017-05-05 2018-11-08 주식회사 유비프로틴 Method for prolonging protein half-life
CN115011579B (en) * 2022-04-28 2024-02-02 北京丹大生物技术有限公司 Taq DNA polymerase mutant and preparation method and application thereof

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5677152A (en) * 1995-08-25 1997-10-14 Roche Molecular Systems, Inc. Nucleic acid amplification using a reersibly inactivated thermostable enzyme
KR20050112686A (en) * 2004-05-27 2005-12-01 바이오퀘스트(주) Chemical-modified thermostable dna polymerase

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB0022458D0 (en) * 2000-09-13 2000-11-01 Medical Res Council Directed evolution method

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5677152A (en) * 1995-08-25 1997-10-14 Roche Molecular Systems, Inc. Nucleic acid amplification using a reersibly inactivated thermostable enzyme
KR20050112686A (en) * 2004-05-27 2005-12-01 바이오퀘스트(주) Chemical-modified thermostable dna polymerase

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
KEOHAVONG ET AL.: "Fidelity of DNA polymerases in DNA amplification", PROC. NATL. ACAD. SCI. USA, vol. 86, 1989, pages 9253 - 9257, XP008126871 *
PAVLOV ET AL.: "Recent developments in the optimization of thermostable DNA polymerases for efficient applications", TRENDS IN BIOTECHNOLOGY, vol. 22, no. 5, May 2004 (2004-05-01), XP004504365 *
SUZUKI ET AL.: "Thermus aquaticus DNA Polymerase I Mutants with Altered Fidelity", THE JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 275, no. 42, 20 October 2000 (2000-10-20), pages 32728 - 32735, XP002369419 *
YOSHIDA ET AL.: "Arg660Ser mutation in Thermus aquaticus DNA polymerase I suppresses T to C transitions: implication of wobble base pair formation at the nucleotide incorporation step", NUCLEIC ACIDS RESEARCH, vol. 29, no. 20, 2001, pages 4206 - 4214, XP008140680 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP7470216B2 (en) 2020-05-29 2024-04-17 ジェノテック コーポレイション DNA polymerase mutants with improved tolerance to genetic mutations

Also Published As

Publication number Publication date
EP2300613A2 (en) 2011-03-30
EP2300613A4 (en) 2011-11-09
WO2009155464A2 (en) 2009-12-23

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