PREPARATION OF CIRCULAR DNA

PURPOSE:To purify a large amount of circular DNA, by the combination of the use of an enzyme digesting straight-chain DNA and inactive to circular DNA and the technique of column chromatography. CONSTITUTION:The cells of higher animal or vegetable are suspended in 50mM trishydrochloric acid, and are...

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Main Authors ANAI MOTOAKI, UMENO MORIO, TSUDA OSAMU, KATOU KAZUTO, FUJIMOTO MASAJI, MAEKAWA NOBUHIKO, TODA MASAAKI, YAMAGISHI HIDEO
Format Patent
LanguageEnglish
Published 07.03.1985
Subjects
BEER
BIOCHEMISTRY
CHEMISTRY
COMPOSITIONS THEREOF
CULTURE MEDIA
ENZYMOLOGY
FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE
METALLURGY
MICROBIOLOGY
MICROORGANISMS OR ENZYMES
MUTATION OR GENETIC ENGINEERING
PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS
SPIRITS
VINEGAR
WINE
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Summary:PURPOSE:To purify a large amount of circular DNA, by the combination of the use of an enzyme digesting straight-chain DNA and inactive to circular DNA and the technique of column chromatography. CONSTITUTION:The cells of higher animal or vegetable are suspended in 50mM trishydrochloric acid, and are dissolved by adding a surface active agent to the suspension. The product is subjected to ultracentrifugation, and the supernatant liquid containing crude extract of the cell is separated. The liquid contains circular DNA, single strand DNA and double strand DNA. The liquid is subjected to ion chromatography to remove single strand DNA, and then treated with ATP-dependent nuclease to digest and depolymerize the double strand DNA. The objective circular DNA can be prepared by this process.
Bibliography:Application Number: JP19830152260