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

Yuzy MATSUO; Hayafumi KISHIMOTO; Tomitaka HORIUCHI; Katsuhiro TANAE; Makoto KAWAMUKAI

doi:10.1271/bbb.90967

Biochemistry & Molecular Biology Communication

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

Yuzy MATSUO, Hayafumi KISHIMOTO, Tomitaka HORIUCHI, Katsuhiro TANAE, Makoto KAWAMUKAI

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Keywords: fission yeast, gap-repair cloning, short tracts of flanking homology, one-step cloning of multiple DNA fragments

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Supplementary material

2010 Volume 74 Issue 3 Pages 685-689

DOI https://doi.org/10.1271/bbb.90967

View "Advance Publication" version (March 7, 2010).

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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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