CN109937252A - 重组dna聚合酶 - Google Patents

重组dna聚合酶 Download PDF

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CN109937252A
CN109937252A CN201780069442.3A CN201780069442A CN109937252A CN 109937252 A CN109937252 A CN 109937252A CN 201780069442 A CN201780069442 A CN 201780069442A CN 109937252 A CN109937252 A CN 109937252A
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methionine
dna polymerase
amino acid
alanine
kod
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CN109937252B (zh
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王林
刘芬
董宇亮
章文蔚
徐崇钧
斯内扎娜·德马纳克
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MGI Tech Co Ltd
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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
    • C12N9/10—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
    • 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)
    • 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
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
    • C07K2319/21—Fusion polypeptide containing a tag with affinity for a non-protein ligand containing a His-tag
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
    • C07K2319/22—Fusion polypeptide containing a tag with affinity for a non-protein ligand containing a Strep-tag
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/40—Fusion polypeptide containing a tag for immunodetection, or an epitope for immunisation
    • C07K2319/41—Fusion polypeptide containing a tag for immunodetection, or an epitope for immunisation containing a Myc-tag
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/40—Fusion polypeptide containing a tag for immunodetection, or an epitope for immunisation
    • C07K2319/43—Fusion polypeptide containing a tag for immunodetection, or an epitope for immunisation containing a FLAG-tag

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
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  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
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  • Biotechnology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
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  • Immunology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Peptides Or Proteins (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

提供一种重组DNA聚合酶，其为如下A)‑C)中任一种：A)所示的蛋白为将野生型KOD型DNA聚合酶氨基酸序列中第408、409和485位氨基酸残基进行修饰，且对所述野生型KOD型DNA聚合酶氨基酸序列中第53、59、199、243、526、558、613、641、671、673、674、692和709位这13位中至少一位的氨基酸残基进行修饰，得到具有DNA聚合酶活性的蛋白；B)所示的蛋白为将A)所示蛋白的氨基酸序列C端的自末端起1‑29个氨基酸去除，保留其余氨基酸残基且具有DNA聚合酶活性的由A)衍生的蛋白质；C)所示的蛋白为将A)或B)所示蛋白的氨基酸序列末端添加标签序列且具有DNA聚合酶活性的由A)或B)衍生的蛋白质。

Description

PCT国内申请，说明书已公开。

Claims (14)

  1. PCT国内申请，权利要求书已公开。
CN201780069442.3A 2017-01-09 2017-01-09 重组dna聚合酶 Active CN109937252B (zh)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108130318A (zh) * 2018-02-28 2018-06-08 深圳市草履虫生物科技有限公司 突变型Taq DNA聚合酶、免核酸提取直接PCR扩增的试剂盒及其应用
WO2022082482A1 (zh) * 2020-10-21 2022-04-28 深圳华大生命科学研究院 重组型kod聚合酶

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117802065A (zh) * 2018-09-03 2024-04-02 深圳华大生命科学研究院 聚合活性提高的重组型kod聚合酶
MX2020013347A (es) 2018-10-31 2021-04-28 Illumina Inc Polimerasas, composiciones y metodos de uso.
JP2022511206A (ja) 2018-12-05 2022-01-31 イラミーナ インコーポレーテッド ポリメラーゼ、組成物、および使用方法
EP4349977A1 (en) * 2021-05-26 2024-04-10 BGI Shenzhen Thermostable b-family dna polymerase mutant and application thereof

Citations (2)

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CN102171338A (zh) * 2008-10-10 2011-08-31 东洋纺织株式会社 新型的具有果糖基缬氨酰基组氨酸氧化酶活性的蛋白质及其修饰产物以及其应用
CN103013945A (zh) * 2013-01-11 2013-04-03 华东理工大学 一种环氧水解酶突变体及其基因和应用

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US20110045489A1 (en) * 2008-04-22 2011-02-24 New England Biolabs, Inc. Polymerases for Incorporating Modified Nucleotides
US9506045B2 (en) * 2012-12-13 2016-11-29 Roche Molecular Systems, Inc. DNA polymerases with improved activity
PT3161154T (pt) * 2014-06-27 2020-06-08 Illumina Inc Polimerases modificadas para incorporação melhorada de análogos de nucleotídeos
EP3201355B1 (en) * 2014-09-30 2019-07-31 Illumina, Inc. Modified polymerases for improved incorporation of nucleotide analogues

Patent Citations (2)

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Publication number Priority date Publication date Assignee Title
CN102171338A (zh) * 2008-10-10 2011-08-31 东洋纺织株式会社 新型的具有果糖基缬氨酰基组氨酸氧化酶活性的蛋白质及其修饰产物以及其应用
CN103013945A (zh) * 2013-01-11 2013-04-03 华东理工大学 一种环氧水解酶突变体及其基因和应用

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108130318A (zh) * 2018-02-28 2018-06-08 深圳市草履虫生物科技有限公司 突变型Taq DNA聚合酶、免核酸提取直接PCR扩增的试剂盒及其应用
CN108130318B (zh) * 2018-02-28 2020-07-14 深圳市艾伟迪生物科技有限公司 突变型Taq DNA聚合酶、免核酸提取直接PCR扩增的试剂盒及其应用
WO2022082482A1 (zh) * 2020-10-21 2022-04-28 深圳华大生命科学研究院 重组型kod聚合酶

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CN109937252B (zh) 2023-03-28
EP3613851A4 (en) 2020-07-01
US20190330602A1 (en) 2019-10-31
EP3613851A1 (en) 2020-02-26
WO2018126470A1 (zh) 2018-07-12
US10883091B2 (en) 2021-01-05

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