Simple and effective gap-repair cloning using short tracts of flanking homology in fission yeast.
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.
Matsuo
Yuzy
Y
Department of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science, Shimane University, Matsue, Japan.
Kishimoto
Hayafumi
H
Horiuchi
Tomitaka
T
Tanae
Katsuhiro
K
Kawamukai
Makoto
M
D019677:Artificial Gene Fusion / Q000379:methods*; D003001:Cloning, Molecular / Q000379:methods; D004260:DNA Repair / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D016254:Mutagenesis, Insertional / Q000379:methods; D016297:Mutagenesis, Site-Directed / Q000379:methods; D010957:Plasmids / Q000235:genetics; D012568:Schizosaccharomyces / Q000235:genetics*