AU2020277641B2 - Phi29 DNA polymerase mutants with improved primer recognition - Google Patents

Phi29 DNA polymerase mutants with improved primer recognition

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Publication number
AU2020277641B2
AU2020277641B2 AU2020277641A AU2020277641A AU2020277641B2 AU 2020277641 B2 AU2020277641 B2 AU 2020277641B2 AU 2020277641 A AU2020277641 A AU 2020277641A AU 2020277641 A AU2020277641 A AU 2020277641A AU 2020277641 B2 AU2020277641 B2 AU 2020277641B2
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AU
Australia
Prior art keywords
phi29
lys
dna
dna polymerase
amplification
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Active
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AU2020277641A
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English (en)
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AU2020277641A1 (en
Inventor
Luis Blanco Davila
Angel Joaquin PICHER SERANTES
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4Basebio SL
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4Basebio SL
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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
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/22—Ribonucleases [RNase]; Deoxyribonucleases [DNase]
    • 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
    • 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
    • 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
    • C12Q1/6869—Methods for sequencing
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y207/00—Transferases transferring phosphorus-containing groups (2.7)
    • C12Y207/07—Nucleotidyltransferases (2.7.7)
    • C12Y207/07007—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
    • 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
    • C12Q2521/00—Reaction characterised by the enzymatic activity
    • C12Q2521/10—Nucleotidyl transfering
    • C12Q2521/101—DNA polymerase
    • 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
    • C12Q2533/00—Reactions characterised by the enzymatic reaction principle used
    • C12Q2533/10—Reactions characterised by the enzymatic reaction principle used the purpose being to increase the length of an oligonucleotide strand
    • C12Q2533/101—Primer extension

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Molecular Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Medicinal Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biophysics (AREA)
  • Immunology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)
AU2020277641A 2019-05-17 2020-05-15 Phi29 DNA polymerase mutants with improved primer recognition Active AU2020277641B2 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US201962849252P 2019-05-17 2019-05-17
US62/849,252 2019-05-17
PCT/EP2020/063740 WO2020234200A1 (en) 2019-05-17 2020-05-15 Phi29 dna polymerase mutants with improved primer recognition

Publications (2)

Publication Number Publication Date
AU2020277641A1 AU2020277641A1 (en) 2021-12-09
AU2020277641B2 true AU2020277641B2 (en) 2026-02-19

Family

ID=71016482

Family Applications (1)

Application Number Title Priority Date Filing Date
AU2020277641A Active AU2020277641B2 (en) 2019-05-17 2020-05-15 Phi29 DNA polymerase mutants with improved primer recognition

Country Status (10)

Country Link
US (1) US20220235337A1 (https=)
EP (1) EP3969575B1 (https=)
JP (2) JP2022533366A (https=)
KR (1) KR20220009977A (https=)
CN (1) CN114174502B (https=)
AU (1) AU2020277641B2 (https=)
CA (1) CA3140244A1 (https=)
ES (1) ES3015001T3 (https=)
SG (1) SG11202112620TA (https=)
WO (1) WO2020234200A1 (https=)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113122517B (zh) * 2021-03-24 2023-02-14 深圳清华大学研究院 聚合酶突变体及其应用
WO2024076991A2 (en) * 2022-10-03 2024-04-11 Singular Genomics Systems, Inc. Modified enzymes and uses thereof
EP4665865A1 (en) 2023-02-17 2025-12-24 Anjarium Biosciences AG Methods of making dna molecules and compositions and uses thereof
WO2025036438A1 (zh) 2023-08-15 2025-02-20 上海金斯康生物科技有限公司 预测突变对蛋白酶活性的影响
CN121343954A (zh) * 2024-07-08 2026-01-16 中国科学院宁波材料技术与工程研究所 一种丧失外切酶活性的phi29 DNA聚合酶突变体及其制备方法和应用

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20170159033A1 (en) * 2015-10-27 2017-06-08 Pacific Biosciences Of California, Inc. Methods, systems, and reagents for direct rna sequencing
WO2019019222A1 (zh) * 2017-07-28 2019-01-31 深圳华大智造科技有限公司 提高了热稳定性的Phi29 DNA聚合酶突变体及其应用

Family Cites Families (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5656493A (en) 1985-03-28 1997-08-12 The Perkin-Elmer Corporation System for automated performance of the polymerase chain reaction
WO2009145820A2 (en) * 2008-03-31 2009-12-03 Pacific Biosciences Of California, Inc. Generation of modified polymerases for improved accuracy in single molecule sequencing
ES2628739T3 (es) 2009-10-15 2017-08-03 Ibis Biosciences, Inc. Amplificación por desplazamiento múltiple
KR101190792B1 (ko) * 2010-08-16 2012-10-12 한국과학기술연구원 변형 뉴클레오티드 및 이를 이용한 실시간 중합효소 반응
US9422535B2 (en) * 2013-04-25 2016-08-23 Thermo Fisher Scientific Baltics Uab phi29 DNA polymerase mutants having increased thermostability and processivity
US9587263B2 (en) * 2014-03-26 2017-03-07 General Electric Company Isothermal amplification under low salt condition
TWI788318B (zh) * 2016-12-19 2023-01-01 美商寬騰矽公司 用於定序反應之聚合酶
CN112805372B (zh) * 2018-10-11 2023-10-31 深圳华大生命科学研究院 热稳定性提升的Phi29 DNA聚合酶突变体及其在测序中的应用

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20170159033A1 (en) * 2015-10-27 2017-06-08 Pacific Biosciences Of California, Inc. Methods, systems, and reagents for direct rna sequencing
WO2019019222A1 (zh) * 2017-07-28 2019-01-31 深圳华大智造科技有限公司 提高了热稳定性的Phi29 DNA聚合酶突变体及其应用

Also Published As

Publication number Publication date
CN114174502B (zh) 2025-09-19
JP2022533366A (ja) 2022-07-22
JP2025108504A (ja) 2025-07-23
CN114174502A (zh) 2022-03-11
AU2020277641A1 (en) 2021-12-09
EP3969575C0 (en) 2025-01-01
CA3140244A1 (en) 2020-11-26
KR20220009977A (ko) 2022-01-25
ES3015001T3 (en) 2025-04-28
EP3969575A1 (en) 2022-03-23
US20220235337A1 (en) 2022-07-28
WO2020234200A1 (en) 2020-11-26
SG11202112620TA (en) 2021-12-30
EP3969575B1 (en) 2025-01-01

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