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Induction of Various Mutations during PCRs with Manganese and 8-Hydroxy-dGTP

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タイトル: Induction of Various Mutations during PCRs with Manganese and 8-Hydroxy-dGTP
著者: Kamiya, Hiroyuki 著作を一覧する
Ito, Mana 著作を一覧する
Harashima, Hideyoshi 著作を一覧する
キーワード: mutagenesis PCR
発行日: 2007年 4月
出版者: Pharmaceutical Society of Japan
誌名: Biological & Pharmaceutical Bulletin
巻: 30
号: 4
開始ページ: 842
終了ページ: 844
出版社 DOI: 10.1248/bpb.30.842
抄録: To induce various mutations randomly, PCRs with Mn2+ and with a mutagenic deoxyribonucleotide, 8-hydroxy-dGTP (8-OH-dGTP), were performed. Mutations were induced by deoxyribonucleotide imbalance plus 500mM Mn2+ in the Mn2+ PCR, and the amplified DNA was inserted into a plasmid. The plasmid library obtained from the transformed bacterial cells was then used as the template in the next PCR, which was done with 50 or 100mM 8-OH-dGTP. Four kinds of mutations, A:T→G:C and G:C→A:T transitions and A:T→T:A and A:T→C:G transversions, occurred with similar frequencies. These results suggest that this strategy will be useful in random PCR mutagenesis for the in vitro evolution of nucleic acids and proteins, and for analyses of residues in these biomolecules.
資料タイプ: article
出現コレクション:雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

提供者: 紙谷 浩之


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