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

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Title: Induction of Various Mutations during PCRs with Manganese and 8-Hydroxy-dGTP
Authors: Kamiya, Hiroyuki Browse this author
Ito, Mana Browse this author
Harashima, Hideyoshi Browse this author →KAKEN DB
Keywords: mutagenesis PCR
Issue Date: Apr-2007
Publisher: Pharmaceutical Society of Japan
Journal Title: Biological & Pharmaceutical Bulletin
Volume: 30
Issue: 4
Start Page: 842
End Page: 844
Publisher DOI: 10.1248/bpb.30.842
Abstract: 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.
Type: article
Appears in Collections:薬学研究院 (Faculty of Pharmaceutical Sciences) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 紙谷 浩之

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