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Bioscience, Biotechnology, and Biochemistry Volume 74, 2010 - Issue 3
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Original Articles

Simple and Effective Gap-Repair Cloning Using Short Tracts of Flanking Homology in Fission Yeast

Yuzy MATSUODepartment of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science, Shimane University
,
Hayafumi KISHIMOTODepartment of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science, Shimane University
,
Tomitaka HORIUCHIDepartment of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science, Shimane University
,
Katsuhiro TANAEDepartment of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science, Shimane University
&
Makoto KAWAMUKAIDepartment of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science, Shimane University
Pages 685-689 | Received 28 Dec 2009, Accepted 02 Feb 2010, Published online: 22 May 2014
 
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Abstract

Gap-repair cloning for plasmid construction in budding yeast is very effective and often used. In contrast, the same method is not widely used in fission yeast, because of a shortage of information on it. Here we describe simple and effective gap-repair cloning for plasmid construction using short tracts of flanking homology. By this method, we combined concentrated DNA fragments with short (20 bp) tracts of flanking homology with the marker gene or the pre-existing gene module. In addition, we found that this method can be applied to one-step cloning of multiple DNA fragments to construct a fusion gene.

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Related Research Data

A restriction-free method for gene reconstitution using two single-primer PCRs in parallel to generate compatible cohesive ends
Source: Springer Nature
A Cloning-Free Method for CRISPR/Cas9-Mediated Genome Editing in Fission Yeast.
Source: Genetics Society of America
A simple and efficient seamless DNA cloning method using SLiCE from Escherichia coli laboratory strains and its application to SLiP site-directed mutagenesis
Source: Springer Science and Business Media LLC
Seamless Ligation Cloning Extract (SLiCE) Method Using Cell Lysates from Laboratory Escherichia coli Strains and its Application to SLiP Site-Directed Mutagenesis.
Source: Springer New York

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