One-step random mutagenesis by error-prone rolling circle amplification

被引:55
作者
Fujii, R [1 ]
Kitaoka, M [1 ]
Hayashi, K [1 ]
机构
[1] Natl Food Res Inst, Tsukuba, Ibaraki 3058642, Japan
关键词
D O I
10.1093/nar/gnh147
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In vitro random mutagenesis is a powerful tool for altering properties of enzymes. We describe here a novel random mutagenesis method using rolling circle amplification, named error-prone RCA. This method consists of only one DNA amplification step followed by transformation of the host strain, without treatment with any restriction enzymes or DNA ligases, and results in a randomly mutated plasmid library with 3-4 mutations per kilobase. Specific primers or special equipment, such as a thermal-cycler, are not required. This method permits rapid preparation of randomly mutated plasmid libraries, enabling random mutagenesis to become a more commonly used technique.
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页数:5
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