Bioscience, Biotechnology, and Biochemistry
Online ISSN : 1347-6947
Print ISSN : 0916-8451
Microbiology & Fermentation Technology Notes
A Rapid, Simple, and Effective Method of Constructing a Randomly Mutagenized Plasmid Library Free from Ligation
Takashi KOYANAGIErina YOSHIDAHiromichi MINAMITakane KATAYAMAHidehiko KUMAGAI
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2008 年 72 巻 4 号 p. 1134-1137

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The QuikChange site-directed mutagenesis methodology was applied to constructing a randomly mutagenized plasmid library simply by adding manganese to the reaction mixture. This method is superior to the normally employed Pol I-type polymerase-based error-prone PCR in that (i) it does not require a subsequent ligation reaction, and (ii) there is no accumulation of mutations at the same site. α-Complementation analysis and subsequent sequence analyses of the lacZα genes in the mutated library revealed that the mutations occurred randomly within the target gene and involved all possible base substitutions.
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© 2008 by Japan Society for Bioscience, Biotechnology, and Agrochemistry
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