KR102442802B1 - 식물 내 니코틴의 노르니코틴으로의 변환 감소 - Google Patents

식물 내 니코틴의 노르니코틴으로의 변환 감소 Download PDF

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KR102442802B1
KR102442802B1 KR1020167031693A KR20167031693A KR102442802B1 KR 102442802 B1 KR102442802 B1 KR 102442802B1 KR 1020167031693 A KR1020167031693 A KR 1020167031693A KR 20167031693 A KR20167031693 A KR 20167031693A KR 102442802 B1 KR102442802 B1 KR 102442802B1
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seq
plant
nicotine demethylase
mutations
mutation
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KR20170002444A (ko
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베르나 리즈슐츠렛
시몬 고엡퍼트
루시엔 보벳
니콜라스 시에로
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필립모리스 프로덕츠 에스.에이.
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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
    • 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
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
    • C12N15/8218—Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]
    • 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
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
    • C12N15/8243—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
    • C—CHEMISTRY; METALLURGY
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
    • C12N9/0073—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14) with NADH or NADPH as one donor, and incorporation of one atom of oxygen 1.14.13
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • 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
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
    • C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y114/00—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14)
    • C12Y114/13—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14) with NADH or NADPH as one donor, and incorporation of one atom of oxygen (1.14.13)
    • 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
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/13—Plant traits
    • 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
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
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  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Cell Biology (AREA)
  • Analytical Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Medicinal Chemistry (AREA)
  • Nutrition Science (AREA)
  • Virology (AREA)
  • Botany (AREA)
  • Mycology (AREA)
  • Immunology (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Manufacture Of Tobacco Products (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
KR1020167031693A 2014-05-08 2015-05-07 식물 내 니코틴의 노르니코틴으로의 변환 감소 Expired - Fee Related KR102442802B1 (ko)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
EP14167598.3 2014-05-08
EP14167598 2014-05-08
EP14001645 2014-05-09
EP14001645.2 2014-05-09
PCT/EP2015/060124 WO2015169927A1 (en) 2014-05-08 2015-05-07 Reduction of nicotine to nornicotine conversion in plants

Publications (2)

Publication Number Publication Date
KR20170002444A KR20170002444A (ko) 2017-01-06
KR102442802B1 true KR102442802B1 (ko) 2022-09-14

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KR1020167031693A Expired - Fee Related KR102442802B1 (ko) 2014-05-08 2015-05-07 식물 내 니코틴의 노르니코틴으로의 변환 감소

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Country Link
US (1) US10415050B2 (enExample)
EP (1) EP3140404A1 (enExample)
JP (1) JP6871158B2 (enExample)
KR (1) KR102442802B1 (enExample)
CN (1) CN106459923B (enExample)
BR (1) BR112016025234A2 (enExample)
CA (1) CA2944965A1 (enExample)
MX (1) MX2016014635A (enExample)
PH (1) PH12016501718A1 (enExample)
RU (1) RU2733837C2 (enExample)
WO (1) WO2015169927A1 (enExample)

Families Citing this family (28)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106636146B (zh) * 2017-02-16 2019-09-13 云南省烟草农业科学研究院 一种降低尼古丁转化率的cyp82e5基因突变体及其应用
US11046969B2 (en) * 2017-05-24 2021-06-29 Epiplanta Biotech Ltd. Transgenic plant and the method for producing the same
WO2018222667A1 (en) * 2017-05-31 2018-12-06 22Nd Century Limited, Llc Genome editing methods for producing low-nicotine tobacco products
WO2019113360A1 (en) 2017-12-07 2019-06-13 University Of Kentucky Research Foundation bZIP TRANSCRIPTION FACTORS REGULATE CONVERSION OF NICOTINE TO NORNICOTINE
CN108192900B (zh) * 2018-01-16 2021-11-30 西南大学 烟草烟碱去甲基化酶基因cyp82e4的编辑失活载体及其应用
KR102995073B1 (ko) 2018-03-28 2026-07-24 필립모리스 프로덕츠 에스.에이. 식물 내 아미노산 함량 조절
WO2019185699A1 (en) 2018-03-28 2019-10-03 Philip Morris Products S.A. Modulating reducing sugar content in a plant
CN108424921B (zh) * 2018-04-20 2021-04-20 云南省烟草农业科学研究院 一种降低尼古丁转化率的cyp82e10基因错义突变体m594及其应用
BR112021001364A2 (pt) 2018-07-26 2021-04-20 Altria Client Services Llc composições e métodos à base de engenharia de pmt para produzir plantas de tabaco e produtos tendo níveis de alcaloides alterados
GB201812603D0 (en) 2018-08-02 2018-09-19 British American Tobacco Investments Ltd Method
WO2020027970A1 (en) * 2018-08-02 2020-02-06 Antidote Therapeutics, Inc. Novel nicotine degrading enzyme variants
EP3902390A1 (en) * 2018-12-30 2021-11-03 Philip Morris Products S.A. Modulation of nitrate levels in plants via mutation of nitrate reductase
KR20220070305A (ko) 2019-10-01 2022-05-30 필립모리스 프로덕츠 에스.에이. 식물 내 환원당 (inv) 함량 조절
WO2021063863A1 (en) 2019-10-01 2021-04-08 Philip Morris Products S.A. Modulating sugar and amino acid content in a plant (sultr3)
WO2021113337A1 (en) * 2019-12-03 2021-06-10 Altria Client Services Llc Compositions and methods for producing tobacco plants and products having altered alkaloid levels
WO2021221752A2 (en) 2020-02-06 2021-11-04 Trustees Of Boston University High throughput assay for identifying microbial redox enzymes
WO2021158973A1 (en) 2020-02-06 2021-08-12 Trustees Of Boston University Enzyme-based electrochemical nicotine biosensor
BR112022021066A2 (pt) * 2020-04-17 2022-12-06 Japan Tobacco Inc Corpo de planta do gênero nicotiana com baixo teor de alcaloide e método de produção do mesmo
CN111505158B (zh) * 2020-05-09 2022-09-23 云南省烟草农业科学研究院 一种基于孵化处理的尼古丁转化烟株色质快速筛查方法
US20240368615A1 (en) 2021-09-10 2024-11-07 Philip Morris Products S.A. Modulating alkaloid profiles in nicotiana tabacum
CN113980978B (zh) * 2021-11-22 2023-12-26 云南中烟工业有限责任公司 一种烟草烟碱转运蛋白相关的基因及其应用
WO2023117661A1 (en) 2021-12-20 2023-06-29 Philip Morris Products S.A. Increasing anatabine in tobacco leaf by regulating methyl putrescine oxidase
US20250084427A1 (en) 2021-12-21 2025-03-13 Philip Morris Products S.A. Modulation of nicotine production by alteration of nicotinamidase expression or function in plants
CN114540382B (zh) * 2022-02-18 2023-07-18 贵州省烟草科学研究院 烟草镉转运基因NtPLA1及应用
KR20250078994A (ko) 2022-10-13 2025-06-04 필립모리스 프로덕츠 에스.에이. Ntp2 조절에 의한 잎 바이오매스 및 질소 사용 효율 증가
WO2024160860A1 (en) 2023-02-02 2024-08-08 Philip Morris Products S.A. Modulation of genes coding for lysine ketoglutarate reductase
KR20250139371A (ko) 2023-02-02 2025-09-23 필립모리스 프로덕츠 에스.에이. 당 수송체의 조절
WO2025003105A1 (en) 2023-06-29 2025-01-02 Philip Morris Products S.A. Modulation of genes coding for glutamate dehydrogenase

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009064771A2 (en) 2007-11-12 2009-05-22 North Carolina State University Alteration of tobacco alkaloid content through modification of specific cytochrome p450 genes
US20100012137A1 (en) 2006-12-15 2010-01-21 U.S. Smokeless Tobacco Company Tobacco plants having reduced nicotine demethylase activity
WO2011088180A1 (en) 2010-01-15 2011-07-21 North Carolina State University Compositions and methods for minimizing nornicotine synthesis in tobacco

Family Cites Families (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5810020A (en) 1993-09-07 1998-09-22 Osmotek, Inc. Process for removing nitrogen-containing anions and tobacco-specific nitrosamines from tobacco products
US6202649B1 (en) 1996-12-02 2001-03-20 Regent Court Technologies Method of treating tobacco to reduce nitrosamine content, and products produced thereby
EE9900590A (et) 1997-06-20 2000-08-15 Regent Court Technologies Nitrosamiinide sisaldust vähendav tubaka töötlemise meetod ja seejuures saadavad tooted
US7700851B2 (en) * 2001-11-13 2010-04-20 U.S. Smokeless Tobacco Company Tobacco nicotine demethylase genomic clone and uses thereof
ATE530656T1 (de) 2005-02-23 2011-11-15 Univ North Carolina State Veränderung des alkaloidgehaltes in tabak durch modifikation spezifischer cytochrome p450 gene.
US7884263B2 (en) * 2005-02-23 2011-02-08 North Carolina State University Alteration of tobacco alkaloid content through modification of specific cytochrome P450 genes
AU2007329776B2 (en) 2006-10-13 2013-05-16 North Carolina State University Alteration of tobacco alkaloid content through modification of specific cytochrome P450 genes
RU2324737C1 (ru) * 2006-10-18 2008-05-20 Институт цитологии и генетики Сибирского отделения Российской академии наук (СО РАН) Способ получения трансгенных растений табака с повышенным содержанием пролина
AP3226A (en) 2007-12-13 2015-04-30 Philip Morris Products Sa Transgenic plants modifier for reducede cadmium transport, derivative products, and related methods
US7883371B1 (en) 2009-07-22 2011-02-08 Hon Hai Precision Ind. Co., Ltd. Electrical connector with improved contact footprints
ES2698154T3 (es) 2010-09-03 2019-01-31 Philip Morris Products Sa Reducción de metales pesados en plantas

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100012137A1 (en) 2006-12-15 2010-01-21 U.S. Smokeless Tobacco Company Tobacco plants having reduced nicotine demethylase activity
WO2009064771A2 (en) 2007-11-12 2009-05-22 North Carolina State University Alteration of tobacco alkaloid content through modification of specific cytochrome p450 genes
WO2011088180A1 (en) 2010-01-15 2011-07-21 North Carolina State University Compositions and methods for minimizing nornicotine synthesis in tobacco

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US20170130239A1 (en) 2017-05-11
BR112016025234A2 (pt) 2017-12-12
CA2944965A1 (en) 2015-11-12
CN106459923B (zh) 2020-08-11
MX2016014635A (es) 2017-02-23
EP3140404A1 (en) 2017-03-15
WO2015169927A1 (en) 2015-11-12
JP2017519519A (ja) 2017-07-20
RU2733837C2 (ru) 2020-10-07
PH12016501718A1 (en) 2016-11-07
KR20170002444A (ko) 2017-01-06
RU2016144409A (ru) 2018-06-13
JP6871158B2 (ja) 2021-05-12
US10415050B2 (en) 2019-09-17
CN106459923A (zh) 2017-02-22
RU2016144409A3 (enExample) 2018-11-29

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