EP1504091A4 - Error-prone dna polymerase i mutants and methods for targeted random mutagenesis in continuous culture using error-prone dna polymerase i mutants - Google Patents

Error-prone dna polymerase i mutants and methods for targeted random mutagenesis in continuous culture using error-prone dna polymerase i mutants

Info

Publication number
EP1504091A4
EP1504091A4 EP03736735A EP03736735A EP1504091A4 EP 1504091 A4 EP1504091 A4 EP 1504091A4 EP 03736735 A EP03736735 A EP 03736735A EP 03736735 A EP03736735 A EP 03736735A EP 1504091 A4 EP1504091 A4 EP 1504091A4
Authority
EP
European Patent Office
Prior art keywords
mutants
error
dna polymerase
prone dna
methods
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.)
Withdrawn
Application number
EP03736735A
Other languages
German (de)
French (fr)
Other versions
EP1504091A2 (en
Inventor
Manuel Camps
Lawrence A Loeb
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
University of Washington
Original Assignee
University of Washington
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 University of Washington filed Critical University of Washington
Publication of EP1504091A2 publication Critical patent/EP1504091A2/en
Publication of EP1504091A4 publication Critical patent/EP1504091A4/en
Withdrawn legal-status Critical Current

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
    • 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
    • C12N15/102—Mutagenizing nucleic acids
    • C12N15/1024—In vivo mutagenesis using high mutation rate "mutator" host strains by inserting genetic material, e.g. encoding an error prone polymerase, disrupting a gene for mismatch repair
    • 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

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • Plant Pathology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
EP03736735A 2002-05-31 2003-05-29 Error-prone dna polymerase i mutants and methods for targeted random mutagenesis in continuous culture using error-prone dna polymerase i mutants Withdrawn EP1504091A4 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US38494402P 2002-05-31 2002-05-31
US384944P 2002-05-31
PCT/US2003/016798 WO2003102213A2 (en) 2002-05-31 2003-05-29 Error-prone dna polymerase i mutants and methods for targeted random mutagenesis in continuous culture using error-prone dna polymerase i mutants

Publications (2)

Publication Number Publication Date
EP1504091A2 EP1504091A2 (en) 2005-02-09
EP1504091A4 true EP1504091A4 (en) 2006-04-26

Family

ID=29712109

Family Applications (1)

Application Number Title Priority Date Filing Date
EP03736735A Withdrawn EP1504091A4 (en) 2002-05-31 2003-05-29 Error-prone dna polymerase i mutants and methods for targeted random mutagenesis in continuous culture using error-prone dna polymerase i mutants

Country Status (5)

Country Link
EP (1) EP1504091A4 (en)
JP (1) JP2005528114A (en)
AU (1) AU2003237269A1 (en)
CA (1) CA2485203A1 (en)
WO (1) WO2003102213A2 (en)

Families Citing this family (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102004005885A1 (en) * 2004-02-05 2005-10-13 Rheinische Friedrich-Wilhelms-Universität Bonn Mutated DNA polymerases with increased mismatch discrimination
EP1926816A1 (en) * 2005-09-21 2008-06-04 Meyer, Andreas In vivo generation of dna, rna, peptide, and protein libraries
US8603797B2 (en) 2010-03-17 2013-12-10 Cornell University Methods and compositions for targeted mutagenesis in bacteria
WO2012139748A1 (en) * 2011-04-11 2012-10-18 Roche Diagnostics Gmbh Dna polymerases with improved activity
EP2788480B1 (en) 2011-12-08 2019-01-16 Roche Diagnostics GmbH Dna polymerases with improved activity
ES2569723T3 (en) 2011-12-08 2016-05-12 F. Hoffmann-La Roche Ag DNA polymerases with enhanced activity
WO2013083264A1 (en) 2011-12-08 2013-06-13 Roche Diagnostics Gmbh Dna polymerases with improved activity
US10760071B2 (en) 2015-07-24 2020-09-01 The Regents Of The University Of California System for continuous mutagenesis in vivo to facilitate directed evolution
CN108130318B (en) * 2018-02-28 2020-07-14 深圳市艾伟迪生物科技有限公司 Mutant Taq DNA polymerase, kit for direct PCR amplification without nucleic acid extraction and application thereof
CN112941155B (en) * 2018-05-14 2023-07-14 北京艾克伦医疗科技有限公司 DNA primer pair with stem-loop structure and application thereof

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0655506A1 (en) * 1994-10-17 1995-05-31 President And Fellows Of Harvard College DNA polymerases having modified nucleotide binding site for DNA sequencing
WO1998035060A1 (en) * 1997-02-07 1998-08-13 Life Technologies, Inc. Polymerases for analyzing or typing polymorphic nucleic acid fragments and uses thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0655506A1 (en) * 1994-10-17 1995-05-31 President And Fellows Of Harvard College DNA polymerases having modified nucleotide binding site for DNA sequencing
WO1998035060A1 (en) * 1997-02-07 1998-08-13 Life Technologies, Inc. Polymerases for analyzing or typing polymorphic nucleic acid fragments and uses thereof

Non-Patent Citations (10)

* Cited by examiner, † Cited by third party
Title
BELL J B ET AL: "Base miscoding and strand misalignment errors by mutator Klenow polymerases with amino acid substitutions at tyrosine 766 in the O helix of the fingers subdomain", JOURNAL OF BIOLOGICAL CHEMISTRY 1997 UNITED STATES, vol. 272, no. 11, 1997, pages 7345 - 7351, XP002369420, ISSN: 0021-9258 *
BLONDAL T ET AL: "CLONING, SEQUENCE ANALYSIS AND FUNCTIONAL CHARACTERIZATION OF DNA POLYMERASE I FROM THE THERMOPHILIC EUBACTERIUM RHODOTHERMUS MARINUS", BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY, ACADEMIC PRESS, US, vol. 34, no. 1, August 2001 (2001-08-01), pages 37 - 45, XP009061567, ISSN: 0885-4513 *
CAMPS M ET AL: "When Pol I goes into high gear: Processive DNA synthesis by Pol I in the cell", CELL CYCLE 2004 UNITED STATES, vol. 3, no. 2, 2004, pages 116 - 118, XP002369425, ISSN: 1538-4101 *
CAMPS MANEL ET AL: "Targeted gene evolution in Escherichia coli using a highly error-prone DNA polymerase I.", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, vol. 100, no. 17, 19 August 2003 (2003-08-19), pages 9727 - 9732, XP002369424, ISSN: 0027-8424 *
DERBYSHIRE V ET AL: "The 3'-5' exonuclease of DNA polymerase I of Escherichia coli: contribution of each amino acid at the active site to the reaction.", THE EMBO JOURNAL. JAN 1991, vol. 10, no. 1, January 1991 (1991-01-01), pages 17 - 24, XP002369423, ISSN: 0261-4189 *
KAUSHIK N ET AL: "Significance of the O-helix residues of Escherichia coli DNA polymerase I in DNA synthesis: dynamics of the dNTP binding pocket.", BIOCHEMISTRY. 4 JUN 1996, vol. 35, no. 22, 4 June 1996 (1996-06-04), pages 7256 - 7266, XP002369422, ISSN: 0006-2960 *
PATEL P H ET AL: "Prokaryotic DNA polymerase I: evolution, structure, and ''base flipping'' mechanism for nucleotide selection", JOURNAL OF MOLECULAR BIOLOGY, LONDON, GB, vol. 308, no. 5, 18 May 2001 (2001-05-18), pages 823 - 837, XP004466165, ISSN: 0022-2836 *
SHINKAI A ET AL: "In vivo mutagenesis by Escherichia coli DNA polymerase I. Ile(709) in motif A functions in base selection.", THE JOURNAL OF BIOLOGICAL CHEMISTRY. 14 DEC 2001, vol. 276, no. 50, 14 December 2001 (2001-12-14), pages 46759 - 46764, XP002369421, ISSN: 0021-9258 *
SUZUKI MOTOSHI ET AL: "Thermus aquaticus DNA polymerase I mutants with altered fidelity. Interacting mutations in the O-helix", JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 275, no. 42, 20 October 2000 (2000-10-20), pages 32728 - 32735, XP002369419, ISSN: 0021-9258 *
TOSAKA AKI ET AL: "O-helix mutant T664P of Thermus aquaticus DNA polymerase I: Altered catalytic properties for incorporation of incorrect nucleotides but not correct nucleotides", JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 276, no. 29, 20 July 2001 (2001-07-20), pages 27562 - 27567, XP002369418, ISSN: 0021-9258 *

Also Published As

Publication number Publication date
CA2485203A1 (en) 2003-12-11
WO2003102213A2 (en) 2003-12-11
JP2005528114A (en) 2005-09-22
EP1504091A2 (en) 2005-02-09
WO2003102213A3 (en) 2004-06-17
AU2003237269A1 (en) 2003-12-19

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