A piggyBac transposon-based mutagenesis system for the fission yeast Schizosaccharomyces pombe.
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.
National Institute of Biological Sciences, Beijing 102206, China.
Jun
J
Li
Jia-Min
JM
Zhang
Xin
X
Li
Fang
F
Suo
Mei-Jun
MJ
Zhang
Wenru
W
Hou
Jinghua
J
Han
Li-Lin
LL
Du
D000483:Alleles; D004251:DNA Transposable Elements; D005819:Genetic Markers; D016681:Genome, Fungal; D016296:Mutagenesis; D010641:Phenotype; D012568:Schizosaccharomyces / Q000235:genetics*
D004251:DNA Transposable Elements; D005819:Genetic Markers
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