METHYLATING ENZYME AND METHYLATION ANALYSIS METHOD USING THE SAME

• Main Authors: MAEDA SHOTA, IIDA YASUHIRO
• Format: Patent
• Language: English; Japanese
• Published: 31.08.2020
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

To provide maintenance methyl transferases that hardly cause de novo activity, and to provide methylation analysis methods which hardly cause misrecognition that a hydroxymethylation site is a methylation site, using the same.SOLUTION: The methylating enzyme of the present invention consists of an amino acid sequence in which both the PCNA binding region and RFTS domain are deleted from Dnmt1, and has an activity of maintaining the methylation state of methylated cytosine in the DNA in the DNA replication process. The present invention also provides a method for analyzing the methylation of DNA by applying the activity of this methylating enzyme to the so-called bisulfite method.SELECTED DRAWING: Figure 1 【課題】de novo 活性が生じにくい維持メチルトランスファーゼの提供及びそれを用いたヒドロキシメチル化部位をメチル化部位であるという誤認識が生じにくいメチル化解析方法の提供。【解決手段】本発明のメチル化酵素は、Ｄｎｍｔ１からＰＣＮＡ結合領域及びＲＦＴＳの両ドメインを欠落させたアミノ酸配列からなり、ＤＮＡの複製過程で前記ＤＮＡ中のメチル化されたシトシンのメチル化状態を維持する活性を有する。本発明では、このメチル化酵素の活性をいわゆるバイサルファイト法に応用して、ＤＮＡのメチル化を解析する方法も提供される。【選択図】図１