A set of loxP marker cassettes for cre-mediated multiple gene disruption in Schizosaccharomyces pombe

Tomoko Iwaki, Kaoru Takegawa

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

For functional analysis, the presence of gene families and isoenzymes often makes it necessary to delete more than one gene, while the number of marker genes is limited in Schizosaccharomyces pombe. Here we describe a loxP-flanked ura4+ cassette and Cre recombinase vector for a Cre-loxP-mediated marker removal procedure in S. pombe. This loxP-ura4-loxP cassette can be used for disruption of hmt1+ as a model target gene. We have constructed two vectors which express Cre recombinase under the control of the nmt1 or nmt41 promoter. Excisive recombination at loxP sites in the chromosome was promoted efficiently and accurately when the Cre recombinase was expressed under the control of the nmt41 promoter. In addition, ura4+ could be excised from the genome by Cre recombinase, when a single loxP site was adjacent to ura4. The use of the Cre-loxP system proved to be a practical strategy to excise a marker gene for repeated use in S. pombe.

Original languageEnglish
Pages (from-to)545-550
Number of pages6
JournalBioscience, Biotechnology and Biochemistry
Volume68
Issue number3
DOIs
Publication statusPublished - Mar 1 2004
Externally publishedYes

    Fingerprint

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

Cite this