WO2003002736A3 - A walk-through technique for in vitro recombination of polynucleotide sequences - Google Patents

A walk-through technique for in vitro recombination of polynucleotide sequences Download PDF

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Publication number
WO2003002736A3
WO2003002736A3 PCT/EP2002/007060 EP0207060W WO03002736A3 WO 2003002736 A3 WO2003002736 A3 WO 2003002736A3 EP 0207060 W EP0207060 W EP 0207060W WO 03002736 A3 WO03002736 A3 WO 03002736A3
Authority
WO
WIPO (PCT)
Prior art keywords
walk
polynucleotide sequences
technique
molecules
vitro recombination
Prior art date
Application number
PCT/EP2002/007060
Other languages
French (fr)
Other versions
WO2003002736A2 (en
Inventor
Zhixin Shao
Peter Kratzsch
Rainer Schmuck
Der Eltz Herbert Von
Janet Kenklies
Original Assignee
Roche Diagnostics Gmbh
Hoffmann La Roche
Zhixin Shao
Peter Kratzsch
Rainer Schmuck
Der Eltz Herbert Von
Janet Kenklies
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 Roche Diagnostics Gmbh, Hoffmann La Roche, Zhixin Shao, Peter Kratzsch, Rainer Schmuck, Der Eltz Herbert Von, Janet Kenklies filed Critical Roche Diagnostics Gmbh
Priority to CA002453864A priority Critical patent/CA2453864A1/en
Priority to JP2003509098A priority patent/JP3967319B2/en
Priority to EP02747439A priority patent/EP1404825A2/en
Publication of WO2003002736A2 publication Critical patent/WO2003002736A2/en
Publication of WO2003002736A3 publication Critical patent/WO2003002736A3/en

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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
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0012—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7)
    • C12N9/0026—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7) acting on CH-NH groups of donors (1.5)
    • C12N9/0032—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7) acting on CH-NH groups of donors (1.5) with oxygen as acceptor (1.5.3)
    • C12N9/0034—Sarcosine oxidase (1.5.3.1)
    • 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
    • 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
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)

Abstract

A simple and efficient method for random in vitro mutagenesis and recombination of polynucleotide sequences based on walk-through with chain-elongating molecules and chain terminating molecules e.g. dNTPs/ddNTPs followed by reassembly and amplification is described. The utility of this method for improving protein structure and/or function is demonstrated by creating novel sarcosine oxidase variants and by recombining human placental alkaline phosphates and calf intestinal alkaline phosphatase for better mutants with improved properties.
PCT/EP2002/007060 2001-06-27 2002-06-26 A walk-through technique for in vitro recombination of polynucleotide sequences WO2003002736A2 (en)

Priority Applications (3)

Application Number Priority Date Filing Date Title
CA002453864A CA2453864A1 (en) 2001-06-27 2002-06-26 A walk-through technique for in vitro recombination of polynucleotide sequences
JP2003509098A JP3967319B2 (en) 2001-06-27 2002-06-26 Walk-through technique for in vitro recombination of polynucleotide sequences
EP02747439A EP1404825A2 (en) 2001-06-27 2002-06-26 A WALK-THROUGH TECHNIQUE FOR i IN VITRO /i RECOMBINATION OF POLYNUCLEOTIDE SEQUENCES

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP01115424.2 2001-06-27
EP01115424 2001-06-27

Publications (2)

Publication Number Publication Date
WO2003002736A2 WO2003002736A2 (en) 2003-01-09
WO2003002736A3 true WO2003002736A3 (en) 2003-07-31

Family

ID=8177828

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2002/007060 WO2003002736A2 (en) 2001-06-27 2002-06-26 A walk-through technique for in vitro recombination of polynucleotide sequences

Country Status (4)

Country Link
EP (1) EP1404825A2 (en)
JP (1) JP3967319B2 (en)
CA (1) CA2453864A1 (en)
WO (1) WO2003002736A2 (en)

Families Citing this family (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2464738A4 (en) * 2009-08-12 2013-05-01 Nugen Technologies Inc Methods, compositions, and kits for generating nucleic acid products substantially free of template nucleic acid
CA2852949A1 (en) 2011-10-19 2013-04-25 Nugen Technologies, Inc. Compositions and methods for directional nucleic acid amplification and sequencing
EP3578697B1 (en) 2012-01-26 2024-03-06 Tecan Genomics, Inc. Compositions and methods for targeted nucleic acid sequence enrichment and high efficiency library generation
US9957549B2 (en) 2012-06-18 2018-05-01 Nugen Technologies, Inc. Compositions and methods for negative selection of non-desired nucleic acid sequences
US20150011396A1 (en) 2012-07-09 2015-01-08 Benjamin G. Schroeder Methods for creating directional bisulfite-converted nucleic acid libraries for next generation sequencing
EP2971130A4 (en) 2013-03-15 2016-10-05 Nugen Technologies Inc Sequential sequencing
CN105849264B (en) 2013-11-13 2019-09-27 纽亘技术公司 For identifying the composition and method that repeat sequencing reading
US9745614B2 (en) 2014-02-28 2017-08-29 Nugen Technologies, Inc. Reduced representation bisulfite sequencing with diversity adaptors
SG11201700891SA (en) 2014-08-06 2017-03-30 Nugen Technologies Inc Digital measurements from targeted sequencing
US11099202B2 (en) 2017-10-20 2021-08-24 Tecan Genomics, Inc. Reagent delivery system
CN112041459A (en) * 2018-01-29 2020-12-04 圣祖德儿童研究医院 Nucleic acid amplification method

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5605793A (en) * 1994-02-17 1997-02-25 Affymax Technologies N.V. Methods for in vitro recombination

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5605793A (en) * 1994-02-17 1997-02-25 Affymax Technologies N.V. Methods for in vitro recombination

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
SHAO Z ET AL: "Engineering new functions and altering existing functions", CURRENT OPINION IN STRUCTURAL BIOLOGY, CURRENT BIOLOGY LTD., LONDON, GB, vol. 6, no. 4, August 1996 (1996-08-01), pages 513 - 518, XP002113761, ISSN: 0959-440X *
SHAO ZHIXIN ET AL: "Random-priming in vitro recombination: An effective tool for directed evolution", NUCLEIC ACIDS RESEARCH, OXFORD UNIVERSITY PRESS, SURREY, GB, vol. 26, no. 2, 15 January 1998 (1998-01-15), pages 681 - 683, XP002162556, ISSN: 0305-1048 *

Also Published As

Publication number Publication date
CA2453864A1 (en) 2003-01-09
WO2003002736A2 (en) 2003-01-09
JP2004533259A (en) 2004-11-04
JP3967319B2 (en) 2007-08-29
EP1404825A2 (en) 2004-04-07

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