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1. (WO2018159669) METHOD FOR REPLICATING OR AMPLIFYING CIRCULAR DNA

Pub. No.:    WO/2018/159669    International Application No.:    PCT/JP2018/007485
Publication Date: Sat Sep 08 01:59:59 CEST 2018 International Filing Date: Thu Mar 01 00:59:59 CET 2018
IPC: C12N 15/10
Applicants: JAPAN SCIENCE AND TECHNOLOGY AGENCY
国立研究開発法人 科学技術振興機構
Inventors: SU'ETSUGU, Masayuki
末次 正幸
NARA, Seia
奈良 聖亜
Title: METHOD FOR REPLICATING OR AMPLIFYING CIRCULAR DNA
Abstract:
Provided is a method that enables replication or amplification of circular DNA, particularly long-chain circular DNA, in a cell-free system. Specifically, provided is a method for inhibiting generation of a DNA multimer which is a by-product, during replication or amplification of circular DNA having a replication initiation sequence (origin of chromosome (oriC)) by using the following enzymes: (1) first enzymes which catalyze replication of the circular DNA; (2) second enzymes which catalyze an Okazaki fragment linking reaction, to synthesize two sister circular DNA molecules forming a catenane; and (3) third enzymes which catalyze a separation reaction of the two sister circular DNA molecules. Further provided is a method that comprises introduction of oriC into circular DNA by using a transposon.