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Nucleic Acids Res. 2011 Mar;39(6):e40. doi: 10.1093/nar/gkq1358. Epub 2011 Jan 18.

A piggyBac transposon-based mutagenesis system for the fission yeast Schizosaccharomyces pombe.

Author information

1
National Institute of Biological Sciences, Beijing 102206, China.

Abstract

The TTAA-specific transposon piggyBac (PB), originally isolated from the cabbage looper moth, Trichoplusia ni, has been utilized as an insertional mutagenesis tool in various eukaryotic organisms. Here, we show that PB transposes in the fission yeast Schizosaccharomyces pombe and leaves almost no footprints. We developed a PB-based mutagenesis system for S. pombe by constructing a strain with a selectable transposon excision marker and an integrated transposase gene. PB transposition in this strain has low chromosomal distribution bias as shown by deep sequencing-based insertion site mapping. Using this system, we obtained loss-of-function alleles of klp5 and klp6, and a gain-of-function allele of dam1 from a screen for mutants resistant to the microtubule-destabilizing drug thiabendazole. From another screen for cdc25-22 suppressors, we obtained multiple alleles of wee1 as expected. The success of these two screens demonstrated the usefulness of this PB-mediated mutagenesis tool for fission yeast.

PMID:
21247877
PMCID:
PMC3064801
DOI:
10.1093/nar/gkq1358
[Indexed for MEDLINE]
Free PMC Article

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