JP2019022513 METHODS FOR PREPARING DNA UNIT COMPOSITIONS AND METHODS FOR CREATING CONCATENATED DNA

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Japan

Application Number 2018192729

Application Date 11.10.2018

Publication Number 2019022513

Publication Date 14.02.2019

Grant Number 6692873

Grant Date 17.04.2020

Publication Kind B2

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Applicants SYNPLOGEN INC
株式会社シンプロジェン

Inventors TSUGE KENJI
柘植　謙爾
ITAYA MITSUYASU
板谷　光泰

Agents 正林　真之
林　一好

Priority Data 2014008690 21.01.2014 JP

Title

(EN) METHODS FOR PREPARING DNA UNIT COMPOSITIONS AND METHODS FOR CREATING CONCATENATED DNA

(JA) 単位ＤＮＡ組成物の調製方法及びＤＮＡ連結体の作製方法

Abstract

(EN)

PROBLEM TO BE SOLVED: To provide a method for preparing a DNA unit composition in which the molar numbers of a plurality of DNA units are more uniform.

SOLUTION: The invention provides a method for preparing a DNA unit composition comprising the steps of: providing solution each containing a respective DNA unit linked to an additional sequence; and determining the concentration of respective DNA unit with the additional sequence in the solution, and on the basis of results thereof, fractionating each of the solution and making the molar number of the DNA unit in each of the solutions closer to being identical to one another. The invention also provides a method for preparing a DNA concatemer comprising the steps of: preparing a DNA unit composition; providing a vector DNA; removing each additional sequence from the DNA unit linked to the additional sequence in the prepared solution using a restriction enzyme; and linking the vector DNA and each unit DNA after the removal step.

SELECTED DRAWING: None

(JA)

【課題】複数の単位ＤＮＡのモル数がより揃えられた単位ＤＮＡ組成物の調製方法及びＤＮＡ連結体の作製方法の提供。
【解決手段】付加配列が連結された複数の単位ＤＮＡを該単位ＤＮＡの種類毎に含む溶液を準備する工程と、各々の溶液の準備後、単位ＤＮＡに付加配列が連結された状態で、各々の溶液中の単位ＤＮＡの濃度を測定し、その結果に基づいて、各々の溶液を分取して、各々の溶液中の単位ＤＮＡのモル数を互いに同一に近づける工程とを有する単位ＤＮＡ組成物の調製方法、及び単位ＤＮＡ組成物を調製する工程と、ベクターＤＮＡを準備する工程と、調製後の溶液中の付加配列が連結された単位ＤＮＡから制限酵素を用いて各々の付加配列を除去する工程と、除去工程後、ベクターＤＮＡ及び各々の単位ＤＮＡを互いに連結する工程とを有するＤＮＡ連結体の作製方法。
【選択図】なし

C	CHEMISTRY; METALLURGY
12	BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N	MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15	Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09	Recombinant DNA-technology
10	Processes for the isolation, preparation or purification of DNA or RNA

C	CHEMISTRY; METALLURGY
12	BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N	MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15	Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09	Recombinant DNA-technology
10	Processes for the isolation, preparation or purification of DNA or RNA

C	CHEMISTRY; METALLURGY
12	BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N	MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15	Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09	Recombinant DNA-technology
10	Processes for the isolation, preparation or purification of DNA or RNA

C	CHEMISTRY; METALLURGY
12	BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N	MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15	Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09	Recombinant DNA-technology
63	Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
64	General methods for preparing the vector, for introducing it into the cell or for selecting the vector-containing host

C	CHEMISTRY; METALLURGY
12	BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N	MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15	Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09	Recombinant DNA-technology
63	Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
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

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1. JP2019022513 - METHODS FOR PREPARING DNA UNIT COMPOSITIONS AND METHODS FOR CREATING CONCATENATED DNA