ArticleNucleic acids research1997
PCR-mediated direct gene disruption in Schizosaccharomyces pombe.
Article in Nucleic acids research, 1997. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed.
- Article
- New and Redesigned pRS Plasmid Shuttle Vectors for Genetic Manipulation of Saccharomycescerevisiae.G3 (Bethesda, Md.) · 2012Article
- Deletion of the CgTPI gene encoding triose phosphate isomerase of Candida glycerinogenes inhibits the biosynthesis of glycerol.Current microbiology · 2007Article
- Six new amino acid-auxotrophic markers for targeted gene integration and disruption in fission yeast.Current genetics · 2007Article
- Schizosaccharomyces pombe essential genes: a pilot study.Genome research · 2003Article
- Involvement of rhp23, a Schizosaccharomyces pombe homolog of the human HHR23A and Saccharomyces cerevisiae RAD23 nucleotide excision repair genes, in cell cycle control and protein ubiquitination.Nucleic acids research · 2002Article
- Insertional mutagenesis based on illegitimate recombination in Schizosaccharomyces pombe.Nucleic acids research · 2000Article
- High-efficiency gene targeting in Schizosaccharomyces pombe using a modular, PCR-based approach with long tracts of flanking homology.Yeast (Chichester, England) · 1999Article
Corrections and comments
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Authors and funding
3 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We have examined the feasibility and efficiency of PCR-mediated direct gene disruptions in the fission yeast Schizosaccharomyces pombe. In the present study, the S.pombe ura4+ gene was amplified by PCR with oligonucleotides that had short flanking regions ( approximately 40 bp) to the target gene. Using this purified PCR product we were able to disrupt genes in an S. pombe strain bearing aura4 deletion, with an efficiency ranging between 1 and 3% among selected transformants. The results indicated that despite S.pombe's preference for non-homologous or illegitimate recombination, even very short stretches of homologous regions could be used to target genes at a defined frequency in this organism. The successful disruption of four independent genes (sts1+, gcs1+, gsh2+and hmt1+) by this method further demonstrates that, despite the relatively low efficiency, the method is very feasible, and it's simplicity, especially when coupled to phenotype-based screening, should greatly facilitate disruption of genes in S.pombe.
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Registered trials
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