CN115485374A - 一种具有高扩增活性的耐热dna聚合酶突变体 - Google Patents

一种具有高扩增活性的耐热dna聚合酶突变体 Download PDF

Info

Publication number
CN115485374A
CN115485374A CN202080100330.1A CN202080100330A CN115485374A CN 115485374 A CN115485374 A CN 115485374A CN 202080100330 A CN202080100330 A CN 202080100330A CN 115485374 A CN115485374 A CN 115485374A
Authority
CN
China
Prior art keywords
dna polymerase
mutant
taq enzyme
mutation
pcr
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.)
Pending
Application number
CN202080100330.1A
Other languages
English (en)
Inventor
蒋析文
刘霭珊
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.)
Guangzhou Da'an Gene Co ltd
Original Assignee
Guangzhou Da'an Gene Co ltd
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 Guangzhou Da'an Gene Co ltd filed Critical Guangzhou Da'an Gene Co ltd
Publication of CN115485374A publication Critical patent/CN115485374A/zh
Pending 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
    • 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
    • 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
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1058—Directional evolution of libraries, e.g. evolution of libraries is achieved by mutagenesis and screening or selection of mixed population of organisms
    • 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
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/70—Vectors or expression systems specially adapted for E. coli
    • 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
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/04—Libraries containing only organic compounds
    • C40B40/06—Libraries containing nucleotides or polynucleotides, or derivatives thereof
    • C40B40/08—Libraries containing RNA or DNA which encodes proteins, e.g. gene libraries

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Medicinal Chemistry (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

一种具有高扩增活性的耐热DNA聚合酶突变体，应用蛋白质定向进化技术，对针对Taq酶的聚合酶活性结构域构建随机突变库，通过逐步添加筛选压力，使不适合的突变自然淘汰，具有优势性状的突变逐步积累，最终筛选出一系列对Taq酶扩增性能、聚合性能有关键作用的氨基酸位点及其突变，并得到高扩增性能的Taq酶突变体。

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。
CN202080100330.1A 2020-04-30 2020-04-30 一种具有高扩增活性的耐热dna聚合酶突变体 Pending CN115485374A (zh)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2020/088341 WO2021217597A1 (zh) 2020-04-30 2020-04-30 一种具有高扩增活性的耐热dna聚合酶突变体

Publications (1)

Publication Number Publication Date
CN115485374A true CN115485374A (zh) 2022-12-16

Family

ID=78373147

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202080100330.1A Pending CN115485374A (zh) 2020-04-30 2020-04-30 一种具有高扩增活性的耐热dna聚合酶突变体

Country Status (4)

Country Link
US (1) US20220325259A1 (zh)
EP (1) EP3922718A4 (zh)
CN (1) CN115485374A (zh)
WO (1) WO2021217597A1 (zh)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024000364A1 (zh) * 2022-06-30 2024-01-04 江南大学 一种谷氨酸棒杆菌高效突变体及重组菌构建方法与应用

Family Cites Families (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050112637A1 (en) * 1995-09-08 2005-05-26 Chatterjee Deb K. Mutant DNA polymerases and uses thereof
US8962293B2 (en) * 2006-10-18 2015-02-24 Roche Molecular Systems, Inc. DNA polymerases and related methods
US8026091B2 (en) * 2007-07-13 2011-09-27 Roche Molecular Systems, Inc. DNA polymerases and related methods
EP2524036B1 (en) * 2009-07-31 2019-06-12 Agilent Technologies, Inc. Thermostable type-a dna polymerase mutants with increased polymerization rate and resistance to inhibitors
US11091745B2 (en) * 2015-05-12 2021-08-17 Dna Polymerase Technology, Inc. Mutant polymerases and uses thereof
US10793841B2 (en) * 2017-06-30 2020-10-06 Codexis, Inc. T7 RNA polymerase variants
CN107475216B (zh) * 2017-08-15 2022-06-14 北京擎科生物科技有限公司 重组型耐热dna聚合酶及其应用
CN109957557B (zh) * 2017-12-26 2023-01-06 广州市锐博生物科技有限公司 Dna聚合酶及其制备方法
WO2019143622A1 (en) * 2018-01-19 2019-07-25 Bio-Rad Laboratories, Inc. Mutant dna polymerases
CN109486788B (zh) * 2018-10-26 2021-10-22 南京市胸科医院 一种突变体dna聚合酶及其制备方法和应用

Also Published As

Publication number Publication date
US20220325259A1 (en) 2022-10-13
EP3922718A4 (en) 2022-03-23
EP3922718A1 (en) 2021-12-15
WO2021217597A1 (zh) 2021-11-04

Similar Documents

Publication Publication Date Title
CN111484987B (zh) 一种具有高扩增活性的耐热dna聚合酶突变体
CN112795551B (zh) 一种耐高温逆转录酶突变体及其应用
CN112795546B (zh) 一种具有高逆转录效率的耐高温逆转录酶突变体及其应用
CN112795548A (zh) 具有高逆转录效率的耐高温逆转录酶突变体及其应用
CN115485374A (zh) 一种具有高扩增活性的耐热dna聚合酶突变体
CN114480334A (zh) 用于新冠病毒检测的逆转录酶突变体
CN112795549A (zh) 一种逆转录酶突变体
CN112779238B (zh) 用于丙肝病毒检测的dna聚合酶混合物
CN112592905B (zh) 用于新型冠状病毒检测的dna聚合酶混合物
CN112779237B (zh) 用于乙肝病毒检测的dna聚合酶混合物
CN114480330A (zh) 多突变位点的逆转录酶突变体
CN114480338A (zh) 用于核酸检测的逆转录酶突变体
CN114480329A (zh) 高效率mmlv酶突变体
CN114480331A (zh) Mmlv酶组合突变体
CN114480337A (zh) 逆转录酶突变体及逆转录方法
CN114480332A (zh) 耐高温m-mlv酶突变体及其应用
CN114480335A (zh) 逆转录酶及逆转录检测试剂
CN114480333A (zh) 逆转录酶突变体及其应用
CN114480336A (zh) 含有的逆转录酶突变体的核酸检测试剂盒
CN112795547A (zh) 高逆转录效率的逆转录酶突变体
CN112795550A (zh) 耐高温逆转录酶突变体

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination