CN102282255B - 多核苷酸装配的组合物和方法 - Google Patents

多核苷酸装配的组合物和方法 Download PDF

Info

Publication number
CN102282255B
CN102282255B CN200980154897.0A CN200980154897A CN102282255B CN 102282255 B CN102282255 B CN 102282255B CN 200980154897 A CN200980154897 A CN 200980154897A CN 102282255 B CN102282255 B CN 102282255B
Authority
CN
China
Prior art keywords
nucleic acid
sequence
restriction enzyme
dna
acid molecule
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.)
Expired - Fee Related
Application number
CN200980154897.0A
Other languages
English (en)
Chinese (zh)
Other versions
CN102282255A (zh
Inventor
扎克·塞贝
雷蒙德·洛
杰弗里·A·尤伯萨克斯
苏尼尔·S·钱德兰
埃里克·杰迪戴亚·迪安
达伦·M·普拉特
肯尼思·俊树·竹冈
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.)
Amyris Inc
Original Assignee
Amyris Inc
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 Amyris Inc filed Critical Amyris Inc
Publication of CN102282255A publication Critical patent/CN102282255A/zh
Application granted granted Critical
Publication of CN102282255B publication Critical patent/CN102282255B/zh
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

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
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1093—General methods of preparing gene libraries, not provided for in other subgroups
    • 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
    • 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
    • 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
    • C12N15/64—General methods for preparing the vector, for introducing it into the cell or for selecting the vector-containing host
    • 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
    • C12N15/66—General methods for inserting a gene into a vector to form a recombinant vector using cleavage and ligation; Use of non-functional linkers or adaptors, e.g. linkers containing the sequence for a restriction endonuclease
    • 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
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B50/00—Methods of creating libraries, e.g. combinatorial synthesis
    • C40B50/06—Biochemical methods, e.g. using enzymes or whole viable microorganisms

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • General Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Bioinformatics & Computational Biology (AREA)
  • Cell Biology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)
CN200980154897.0A 2008-11-19 2009-11-19 多核苷酸装配的组合物和方法 Expired - Fee Related CN102282255B (zh)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
US11610908P 2008-11-19 2008-11-19
US61/116,109 2008-11-19
US16223009P 2009-03-20 2009-03-20
US61/162,230 2009-03-20
PCT/US2009/065048 WO2010059763A1 (en) 2008-11-19 2009-11-19 Compositions and methods for the assembly of polynucleotides

Publications (2)

Publication Number Publication Date
CN102282255A CN102282255A (zh) 2011-12-14
CN102282255B true CN102282255B (zh) 2014-05-07

Family

ID=41698076

Family Applications (1)

Application Number Title Priority Date Filing Date
CN200980154897.0A Expired - Fee Related CN102282255B (zh) 2008-11-19 2009-11-19 多核苷酸装配的组合物和方法

Country Status (12)

Country Link
US (3) US8221982B2 (cg-RX-API-DMAC7.html)
EP (1) EP2358875B1 (cg-RX-API-DMAC7.html)
JP (1) JP2012509084A (cg-RX-API-DMAC7.html)
KR (1) KR101794298B1 (cg-RX-API-DMAC7.html)
CN (1) CN102282255B (cg-RX-API-DMAC7.html)
AU (1) AU2009316660B2 (cg-RX-API-DMAC7.html)
BR (1) BRPI0922187B1 (cg-RX-API-DMAC7.html)
CA (1) CA2744153C (cg-RX-API-DMAC7.html)
MX (1) MX2011005195A (cg-RX-API-DMAC7.html)
SG (1) SG171760A1 (cg-RX-API-DMAC7.html)
WO (1) WO2010059763A1 (cg-RX-API-DMAC7.html)
ZA (1) ZA201103637B (cg-RX-API-DMAC7.html)

Families Citing this family (62)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2358875B1 (en) * 2008-11-19 2015-08-12 Amyris, Inc. Compositions and methods for the assembly of polynucleotides
US7919605B1 (en) 2010-08-30 2011-04-05 Amyris, Inc. Nucleic acids, compositions and methods for the excision of target nucleic acids
SG194089A1 (en) 2011-04-27 2013-11-29 Amyris Inc Methods for genomic modification
SG11201402083XA (en) 2011-11-09 2014-06-27 Amyris Inc Production of acetyl-coenzyme a derived isoprenoids
US8332160B1 (en) 2011-11-17 2012-12-11 Amyris Biotechnologies, Inc. Systems and methods for engineering nucleic acid constructs using scoring techniques
AU2013203380A1 (en) * 2012-01-13 2013-08-01 Chromatin, Inc. Engineering plants with rate limiting farnesene metabolic genes
WO2013144257A1 (en) * 2012-03-27 2013-10-03 Dsm Ip Assets B.V. Cloning method
CN104508130B (zh) * 2012-06-29 2016-09-28 麻省理工学院 大规模并行组合遗传学
US9976162B2 (en) 2012-12-10 2018-05-22 Agilent Technologies, Inc. Pairing code directed assembly
BR112015023089A2 (pt) 2013-03-15 2017-11-21 Amyris Inc célula hospedeira, método para produzir um isoprenoide, e método para aumentar a produção de acetil-coa ou um composto derivado de acetil-coa
US10494645B2 (en) 2013-04-18 2019-12-03 Fondazione Telethon Effective delivery of large genes by dual AAV vectors
MX2016007797A (es) 2013-12-19 2016-09-07 Amyris Inc Metodos para integracion genomica.
WO2015120403A1 (en) 2014-02-08 2015-08-13 The Regents Of The University Of Colorado, A Body Corporate Multiplexed linking pcr
US11898141B2 (en) * 2014-05-27 2024-02-13 The Broad Institute, Inc. High-throughput assembly of genetic elements
US11518797B2 (en) 2014-11-11 2022-12-06 Clara Foods Co. Methods and compositions for egg white protein production
US20180127804A1 (en) 2014-12-05 2018-05-10 Amyris, Inc. High-throughput sequencing of polynucleotides
CN107466325A (zh) * 2015-03-03 2017-12-12 泰莱托恩基金会 多重载体系统及其应用
WO2016201142A1 (en) 2015-06-09 2016-12-15 Life Technologies Corporation Methods, systems, compositions, kits, apparatus and computer-readable media for molecular tagging
US10793851B2 (en) 2015-08-04 2020-10-06 The Regents Of The University Of Colorado, A Body Corporate Multiplexed binary assembly and quantitative tracking of bacterial populations
US11085046B2 (en) 2015-10-29 2021-08-10 Amyris, Inc. Compositions and methods for production of myrcene
CN106811459B (zh) * 2015-11-27 2019-09-27 清华大学 一种利用短链引物阻遏提高dna克隆及组装效率的方法
BR112018011503A2 (pt) 2015-12-07 2018-12-04 Zymergen Inc promotores da corynebacterium glutamicum
US11208649B2 (en) 2015-12-07 2021-12-28 Zymergen Inc. HTP genomic engineering platform
US9988624B2 (en) 2015-12-07 2018-06-05 Zymergen Inc. Microbial strain improvement by a HTP genomic engineering platform
WO2017201311A2 (en) 2016-05-18 2017-11-23 Amyris, Inc. Compositions and methods for genomic integration of nucleic acids into exogenous landing pads
EP3478833A4 (en) 2016-06-30 2019-10-02 Zymergen, Inc. METHOD FOR PRODUCING A BACTERIAL HEMOGLOBIN LIBRARY AND ITS USE
JP2019519241A (ja) 2016-06-30 2019-07-11 ザイマージェン インコーポレイテッド グルコース透過酵素ライブラリーを生成するための方法およびその使用
GB201621589D0 (en) * 2016-12-19 2017-02-01 Univ Gent And Katholieke Univ Leuven K U Leuven R&D Polynucleotide shuffling method
EP3574105B1 (en) 2017-01-25 2021-04-21 Amyris Bio Products Portugal, Unipessoal, LDA Co-production of a sesquiterpene and a carotenoid
LT3645719T (lt) 2017-06-30 2022-05-25 Inscripta, Inc. Automatinio ląstelių apdorojimo būdai, moduliai, priemonės ir sistemos
US10738327B2 (en) 2017-08-28 2020-08-11 Inscripta, Inc. Electroporation cuvettes for automation
EP3684927B1 (en) 2017-09-18 2024-05-29 Amyris, Inc. Methods for genomic integration for kluyveromyces host cells
WO2019055495A1 (en) 2017-09-18 2019-03-21 Amyris, Inc. METHODS FOR GENETICALLY MODIFYING KLUYVEROMYCES HOST CELLS
WO2019055535A1 (en) 2017-09-18 2019-03-21 Amyris, Inc. METHODS OF GENOMIC INTEGRATION FOR HOST CELLS COMPRISING HANSENULA AND ARXULA
WO2019055481A1 (en) 2017-09-18 2019-03-21 Amyris, Inc. METHODS OF GENOMIC INTEGRATION IN PICHIA AND OTHER HOST CELLS
WO2019064242A1 (en) * 2017-09-29 2019-04-04 Victoria Link Limited MODULAR DNA ASSEMBLY SYSTEM
US10443074B2 (en) 2017-09-30 2019-10-15 Inscripta, Inc. Modification of cells by introduction of exogenous material
AU2019241967A1 (en) 2018-03-29 2020-11-19 Inscripta, Inc. Automated control of cell growth rates for induction and transformation
WO2019200004A1 (en) 2018-04-13 2019-10-17 Inscripta, Inc. Automated cell processing instruments comprising reagent cartridges
AU2019256237A1 (en) * 2018-04-17 2020-10-29 Nanjing GenScript Biotech Co., Ltd. DNA construct design system
US10501738B2 (en) 2018-04-24 2019-12-10 Inscripta, Inc. Automated instrumentation for production of peptide libraries
US10858761B2 (en) 2018-04-24 2020-12-08 Inscripta, Inc. Nucleic acid-guided editing of exogenous polynucleotides in heterologous cells
US10526598B2 (en) 2018-04-24 2020-01-07 Inscripta, Inc. Methods for identifying T-cell receptor antigens
US10532324B1 (en) 2018-08-14 2020-01-14 Inscripta, Inc. Instruments, modules, and methods for improved detection of edited sequences in live cells
US11142740B2 (en) 2018-08-14 2021-10-12 Inscripta, Inc. Detection of nuclease edited sequences in automated modules and instruments
US10533152B1 (en) 2018-08-14 2020-01-14 Inscripta, Inc. Instruments, modules, and methods for improved detection of edited sequences in live cells
AU2019363487A1 (en) 2018-08-30 2021-04-15 Inscripta, Inc. Improved detection of nuclease edited sequences in automated modules and instruments
KR20210088615A (ko) * 2018-10-31 2021-07-14 지머젠 인코포레이티드 Dna 라이브러리의 다중 결정적 어셈블리
EA202192652A1 (ru) * 2019-04-01 2022-01-12 Нэшнл Юниверсити Корпорейшн Кобе Юниверсити Способ конструирования химерной плазмидной библиотеки
AU2020297499A1 (en) 2019-06-21 2022-02-03 Inscripta, Inc. Genome-wide rationally-designed mutations leading to enhanced lysine production in E. coli
US10927385B2 (en) 2019-06-25 2021-02-23 Inscripta, Inc. Increased nucleic-acid guided cell editing in yeast
US12096784B2 (en) 2019-07-11 2024-09-24 Clara Foods Co. Protein compositions and consumable products thereof
CA3146649A1 (en) 2019-07-11 2021-01-14 Clara Foods Co. Protein compositions and consumable products thereof
US10927360B1 (en) 2019-08-07 2021-02-23 Clara Foods Co. Compositions comprising digestive enzymes
US10689669B1 (en) 2020-01-11 2020-06-23 Inscripta, Inc. Automated multi-module cell processing methods, instruments, and systems
JP7473953B2 (ja) * 2020-03-24 2024-04-24 国立大学法人 東京大学 連結ｄｎａの製造方法及びそれに用いるためのベクターの組み合わせ
WO2021211972A1 (en) * 2020-04-16 2021-10-21 Zymergen Inc. Circular-permuted nucleic acids for homology-directed editing
US20210332388A1 (en) 2020-04-24 2021-10-28 Inscripta, Inc. Compositions, methods, modules and instruments for automated nucleic acid-guided nuclease editing in mammalian cells
US11787841B2 (en) 2020-05-19 2023-10-17 Inscripta, Inc. Rationally-designed mutations to the thrA gene for enhanced lysine production in E. coli
CN112813103A (zh) * 2021-01-28 2021-05-18 云舟生物科技（广州）有限公司 一种标准品载体及其在测定重组腺相关病毒滴度中的应用
CN113943666B (zh) * 2021-11-11 2023-08-22 广西壮族自治区农业科学院 一种酿酒酵母工程菌及其应用
CN115261377B (zh) * 2021-12-27 2024-11-19 天津大学 一种dna串联重复单元的拆分组装方法

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030022179A1 (en) * 2000-08-21 2003-01-30 Chesnut Jonathan D. Methods and reagents for molecular cloning
CN1427079A (zh) * 2002-09-30 2003-07-02 华南农业大学 多基因载体的构建方法与应用
WO2007038276A2 (en) * 2005-09-22 2007-04-05 Intrexon Corporation Dna modular cloning vector plasmids and methods for their use
WO2008045380A2 (en) * 2006-10-04 2008-04-17 Codon Devices, Inc. Nucleic acid libraries and their design and assembly

Family Cites Families (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5057419A (en) * 1988-09-22 1991-10-15 Rutgers University Genetically engineered plasmid and organisms for the production of specialized oils
US5976846A (en) * 1996-01-13 1999-11-02 Passmore; Steven E. Method for multifragment in vivo cloning and mutation mapping
CA2303420A1 (en) * 1997-09-29 1999-04-08 City Of Hope Efficient linking of nucleic acid segments
ES2324513T3 (es) * 1999-03-18 2009-08-10 Complete Genomics As Procedimientos de clonacion y produccion de cadenas de fragmentos con contenido de informacion legible.
CA2317865A1 (en) 2000-03-07 2001-09-07 University Of Guelph Dna joining method
US20030087254A1 (en) * 2001-04-05 2003-05-08 Simon Delagrave Methods for the preparation of polynucleotide libraries and identification of library members having desired characteristics
US20020165175A1 (en) * 2001-04-17 2002-11-07 Xiaowu Liang Fast and enzymeless cloning of nucleic acid fragments
DK1929012T3 (da) * 2005-08-11 2011-01-31 Synthetic Genomics Inc Fremgangsmåde til in vitro-rekombination
CA2618699C (en) * 2005-08-11 2012-10-02 J. Craig Venter Institute, Inc. In vitro recombination method
WO2008095927A1 (en) * 2007-02-05 2008-08-14 Philipps-Universität Marburg Method of cloning at least one nucleic acid molecule of interest using type iis restriction endonucleases, and corresponding cloning vectors, kits and system using type iis restriction endonucleases
WO2009048885A2 (en) 2007-10-08 2009-04-16 Synthetic Genomics, Inc. Assembly of large nucleic acids
EP2358875B1 (en) * 2008-11-19 2015-08-12 Amyris, Inc. Compositions and methods for the assembly of polynucleotides

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030022179A1 (en) * 2000-08-21 2003-01-30 Chesnut Jonathan D. Methods and reagents for molecular cloning
CN1427079A (zh) * 2002-09-30 2003-07-02 华南农业大学 多基因载体的构建方法与应用
WO2007038276A2 (en) * 2005-09-22 2007-04-05 Intrexon Corporation Dna modular cloning vector plasmids and methods for their use
WO2008045380A2 (en) * 2006-10-04 2008-04-17 Codon Devices, Inc. Nucleic acid libraries and their design and assembly

Also Published As

Publication number Publication date
KR101794298B1 (ko) 2017-11-20
BRPI0922187B1 (pt) 2021-05-04
US8110360B2 (en) 2012-02-07
US8221982B2 (en) 2012-07-17
US8546136B2 (en) 2013-10-01
US20120245056A1 (en) 2012-09-27
BRPI0922187A2 (pt) 2018-10-23
AU2009316660A1 (en) 2011-07-07
ZA201103637B (en) 2012-08-29
AU2009316660B2 (en) 2015-01-29
WO2010059763A1 (en) 2010-05-27
JP2012509084A (ja) 2012-04-19
US20100136633A1 (en) 2010-06-03
KR20110102351A (ko) 2011-09-16
US20100124768A1 (en) 2010-05-20
EP2358875A1 (en) 2011-08-24
MX2011005195A (es) 2011-09-01
CN102282255A (zh) 2011-12-14
CA2744153C (en) 2020-07-07
SG171760A1 (en) 2011-07-28
EP2358875B1 (en) 2015-08-12
CA2744153A1 (en) 2010-05-27

Similar Documents

Publication Publication Date Title
CN102282255B (zh) 多核苷酸装配的组合物和方法
US8332160B1 (en) Systems and methods for engineering nucleic acid constructs using scoring techniques
EP3257942B1 (en) Method for assembly of polynucleic acid sequences
CN104685116A (zh) 用于核酸组装和高通量测序的方法
CN102016070A (zh) 体外连接和组合装配核酸分子的方法
JP2020519304A (ja) ゲノム大断片のダイレクトクローニングおよびｄｎａマルチ分子構築の新手法
CN102027112B (zh) 核酸重组方法
US20190010530A1 (en) Methods and compositions for nucleic acid assembly
Prins et al. Oligo pools as an affordable source of synthetic DNA for cost-effective library construction of protein variants
WO2022187697A1 (en) In vivo dna assembly and analysis
CN117677694A (zh) 体内dna组装和分析
WO2004011608A2 (en) Methods and nucleic acid vectors for rapid expression and screening of cdna clones
Sylvestre et al. Massive Mutagenesis®: The path to smarter genetic libraries

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20140507

CF01 Termination of patent right due to non-payment of annual fee