Efficient targeted integration at leu1-32 and ura4-294 in Schizosaccharomyces pombe.
Homologous integration into the fission yeast Schizosaccharomyces pombe has not been well characterized. In this study, we have examined integration of plasmids carrying the leu1+ and ura4+ genes into their chromosomal loci. Genomic DNA blot analysis demonstrated that the majority of transformants have one or more copies of the plasmid vector integrated via homologous recombination with a much smaller fraction of gene conversion to leu1+ or ura4+. Non-homologous recombination events were not observed for either gene. We describe the construction of generally useful leu1+ and ura4+ plasmids for targeted integration at the leu1-32 and ura4-294 loci of S. pombe.
Keeney
J B
JB
Johns Hopkins University School of Medicine, Department of Molecular Biology and Genetics, Baltimore, Maryland 21205.
Boeke
J D
JD
D002874:Chromosome Mapping; D004271:DNA, Fungal / Q000235:genetics; D005785:Gene Conversion; D005800:Genes, Fungal; D005822:Genetic Vectors; D011995:Recombination, Genetic; D012568:Schizosaccharomyces / Q000235:genetics*; D014170:Transformation, Genetic
D004271:DNA, Fungal