CU20180158A7 - Método para identificar un mutante dentro de una población de organismos aplicando un enfoque de conjunto y división - Google Patents

Método para identificar un mutante dentro de una población de organismos aplicando un enfoque de conjunto y división

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Publication number
CU20180158A7
CU20180158A7 CU2018000158A CU20180158A CU20180158A7 CU 20180158 A7 CU20180158 A7 CU 20180158A7 CU 2018000158 A CU2018000158 A CU 2018000158A CU 20180158 A CU20180158 A CU 20180158A CU 20180158 A7 CU20180158 A7 CU 20180158A7
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CU
Cuba
Prior art keywords
organisms
dpcr
finding
mutant
population
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CU2018000158A
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English (en)
Inventor
Massimiliano Carciofi
Søren Knudsen
Ole Oslen
Magus Wohlfahrt Rasmessen
Birgitte Skadhauge
Alexander Striebeck
Hanne Cecilie Thomsen
Toni Wendt
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Carlsberg As
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Publication of CU20180158A7 publication Critical patent/CU20180158A7/es

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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
    • C12N15/01—Preparation of mutants without inserting foreign genetic material therein; Screening processes therefor
    • 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
    • C12Q1/686—Polymerase chain reaction [PCR]
    • 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
    • C12Q1/6876—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
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
    • A01H6/46—Gramineae or Poaceae, e.g. ryegrass, rice, wheat or maize
    • A01H6/4624—Hordeum vulgarus [barley]
    • 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
    • C12N15/102—Mutagenizing nucleic acids
    • C12N15/1024—In vivo mutagenesis using high mutation rate "mutator" host strains by inserting genetic material, e.g. encoding an error prone polymerase, disrupting a gene for mismatch repair
    • 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
    • C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
    • 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
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6827—Hybridisation assays for detection of mutation or polymorphism
    • 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
    • C12Q1/6858—Allele-specific amplification
    • 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
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • 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
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B30/00—Methods of screening libraries
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/02—Libraries contained in or displayed by microorganisms, e.g. bacteria or animal cells; Libraries contained in or displayed by vectors, e.g. plasmids; Libraries containing only microorganisms or vectors

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Biomedical Technology (AREA)
  • Analytical Chemistry (AREA)
  • Immunology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Plant Pathology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Developmental Biology & Embryology (AREA)
  • Botany (AREA)
  • Environmental Sciences (AREA)
  • Physiology (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Cell Biology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Mycology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

<p>En los enfoques de mejoramiento de plantas tradicionales, puede utilizarse mutagénesis química para introducir sustituciones nucleotídicas aleatoriamente en el genoma de una planta, es decir sin posibilidades de controlar los sitios de los cambios nucleotídicos. A causa de las complejidades genómicas, la probabilidad estadística es extremadamente baja en cuanto a hallar una sustitución nucleotídica predeterminada. La presente invención, sin embargo, demuestra cómo se desarrolla un uso novedoso, alternativo, de la reacción en cadena de la polimerasa digital (dPCR), preferiblemente dPCR en gotas (ddPCR), para explotar el hallazgo de sustituciones nucleotídicas específicas en genes mutados. La plataforma completa comprende un método de relevamiento con una biblioteca de organismos mutagenizados, sistemas basados en PCR digital y una configuración para propagar y analizar organismos mutados identificados.</p>
CU2018000158A 2016-07-01 2017-06-23 Método para identificar un mutante dentro de una población de organismos aplicando un enfoque de conjunto y división CU20180158A7 (es)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DKPA201670485 2016-07-01
PCT/EP2017/065516 WO2018001884A1 (en) 2016-07-01 2017-06-23 Method to screen for a mutant within a population of organisms by applying a pooling and splitting approach

Publications (1)

Publication Number Publication Date
CU20180158A7 true CU20180158A7 (es) 2019-08-06

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CU2018000158A CU20180158A7 (es) 2016-07-01 2017-06-23 Método para identificar un mutante dentro de una población de organismos aplicando un enfoque de conjunto y división

Country Status (25)

Country Link
US (2) US11884972B2 (es)
EP (3) EP3478832B1 (es)
JP (2) JP7208797B2 (es)
CN (2) CN114959098A (es)
AR (1) AR110103A1 (es)
AU (3) AU2017288960B2 (es)
BR (1) BR112018076429A2 (es)
CA (1) CA3026016A1 (es)
CL (1) CL2018003710A1 (es)
CO (1) CO2019000828A2 (es)
CR (1) CR20190040A (es)
CU (1) CU20180158A7 (es)
DK (1) DK3478832T3 (es)
EA (1) EA201892809A1 (es)
ES (2) ES2916221T3 (es)
IL (1) IL263451B (es)
MA (3) MA52594A (es)
MX (2) MX2018016110A (es)
PE (1) PE20190228A1 (es)
PL (2) PL3792357T3 (es)
PT (1) PT3478832T (es)
SG (1) SG11201811021YA (es)
UY (1) UY37312A (es)
WO (1) WO2018001884A1 (es)
ZA (3) ZA201808218B (es)

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CN108913698B (zh) * 2018-07-25 2021-10-26 安徽农业大学 一种与小麦穗发芽抗性/感性相关的caps标记及其应用
CN110724741B (zh) * 2019-07-24 2020-05-19 艾普拜生物科技(苏州)有限公司 一种检测微小残留白血病相关融合基因的引物、探针及其试剂盒
EP4013843A1 (en) * 2019-08-16 2022-06-22 Heineken Supply Chain B.V. Production of an alcohol-free beverage
EP3785529A1 (en) 2019-08-28 2021-03-03 Carlsberg A/S Barley plants with altered protein content in grains
EP4040946A1 (en) * 2019-10-10 2022-08-17 Carlsberg A/S Methods for preparing mutant plants
US20240043911A1 (en) * 2020-02-06 2024-02-08 The Administrators Of The Tulane Educational Fund Crispr-based assay for detecting tb in bodily fluids
MX2022010777A (es) 2020-03-02 2022-11-30 Carlsberg As Plantas de cebada con elevada actividad de dextrinasa limite.
EP4029935A1 (en) 2021-01-14 2022-07-20 Carlsberg A/S S. eubayanus strain and hybrids thereof
CA3234121A1 (en) * 2021-10-25 2023-05-04 Sarah KANE Off-target blocking sequences to improve target discrimination by polymerase chain reaction
CA3240580A1 (en) 2021-12-22 2023-06-29 Timothy James Golds Improved screening method for genome edited events
AU2023231447A1 (en) 2022-03-11 2024-09-05 Carlsberg A/S Modulation of protein levels
CN120225050A (zh) 2022-11-16 2025-06-27 哥本哈根大学 种子硫代葡萄糖苷含量低的植物
WO2024133900A1 (en) 2022-12-22 2024-06-27 University Of Copenhagen Improved (-)-sparteine production
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CN117965576B (zh) * 2024-02-01 2024-08-27 北京林业大学 一种王百合苷生物合成基因LlHCT1及其应用
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DK3478832T3 (da) 2022-05-30
CN109477087A (zh) 2019-03-15
IL263451B (en) 2022-08-01
WO2018001884A1 (en) 2018-01-04
EP3693458A1 (en) 2020-08-12
AU2019268102A1 (en) 2019-12-12
JP7208797B2 (ja) 2023-01-19
US12428678B2 (en) 2025-09-30
PT3478832T (pt) 2022-05-19
ZA201907775B (en) 2022-09-28
AR110103A1 (es) 2019-02-27
MA52594A (fr) 2021-03-17
US20190194723A1 (en) 2019-06-27
JP2019524083A (ja) 2019-09-05
AU2017288960B2 (en) 2019-12-12
CL2018003710A1 (es) 2019-03-22
EP3792357B1 (en) 2025-05-07
ZA201907774B (en) 2022-07-27
NZ749382A (en) 2025-06-27
US11884972B2 (en) 2024-01-30
US20240182961A1 (en) 2024-06-06
ES2916221T3 (es) 2022-06-29
CR20190040A (es) 2019-05-09
CN114959098A (zh) 2022-08-30
CO2019000828A2 (es) 2019-04-30
JP2022101703A (ja) 2022-07-06
PL3478832T3 (pl) 2022-06-06
EP3792357A1 (en) 2021-03-17
AU2019268103A1 (en) 2019-12-12
MX2022014965A (es) 2023-01-11
AU2019268103B2 (en) 2020-03-12
MX2018016110A (es) 2019-05-30
PL3792357T3 (pl) 2025-07-07
JP7539428B2 (ja) 2024-08-23
ZA201808218B (en) 2020-02-26
BR112018076429A2 (pt) 2019-10-01
SG11201811021YA (en) 2019-01-30
PE20190228A1 (es) 2019-02-13
EP3478832A1 (en) 2019-05-08
IL263451A (en) 2019-01-31
EA201892809A1 (ru) 2019-06-28
MA54568A (fr) 2021-10-27
CN109477087B (zh) 2022-06-24
EP3478832B1 (en) 2022-02-23
ES3032697T3 (en) 2025-07-23
AU2019268102B2 (en) 2020-03-05
AU2017288960A1 (en) 2019-01-17
UY37312A (es) 2018-02-28
CA3026016A1 (en) 2018-01-04
EP3792357C0 (en) 2025-05-07
MA45531A (fr) 2019-05-08

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