Novel PCR-mediated mutagenesis employing DNA containing a natural abasic site as a template and translesional Taq DNA polymerase

Atsushi Kobayashi; Motomitsu Kitaoka; Kiyoshi Hayashi

doi:10.1016/j.jbiotec.2004.10.016

Novel PCR-mediated mutagenesis employing DNA containing a natural abasic site as a template and translesional Taq DNA polymerase

J Biotechnol. 2005 Mar 30;116(3):227-32. doi: 10.1016/j.jbiotec.2004.10.016. Epub 2004 Dec 25.

Authors

Atsushi Kobayashi¹, Motomitsu Kitaoka, Kiyoshi Hayashi

Affiliation

¹ National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.

PMID: 15707683
DOI: 10.1016/j.jbiotec.2004.10.016

Abstract

We describe a novel method of PCR-mediated mutagenesis employing DNA containing a natural abasic site and translesional Taq DNA polymerase. This method incorporated an adenine (80.8%) or guanine (7.7%) residue or led to a base deletion mutation (11.2%) opposite the abasic site. We conclude that the combination of DNA containing an abasic site and translesional Taq DNA polymerase is an easy and useful technique for PCR-mediated mutagenesis, having advantages different from those of conventional error-prone PCR.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

DNA / genetics*
DNA / metabolism*
DNA Damage
Genetic Engineering / methods*
Mutagenesis, Site-Directed / genetics*
Polymerase Chain Reaction / methods*
Taq Polymerase / metabolism*
Templates, Genetic

Substances

DNA
Taq Polymerase