RRC ID 885
Author Iwaki T, Takegawa K.
Title A set of loxP marker cassettes for Cre-mediated multiple gene disruption in Schizosaccharomyces pombe.
Journal Biosci. Biotechnol. Biochem.
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.
Volume 68(3)
Pages 545-50
Published 2004-3
DOI 10.1271/bbb.68.545
PMID 15056885
MeSH ATP-Binding Cassette Transporters / genetics Base Sequence Cadmium Chloride / toxicity Gene Expression Regulation, Fungal Gene Silencing Genetic Markers Genetic Vectors / genetics* Integrases / genetics* Integrases / metabolism Isoenzymes / genetics Molecular Sequence Data Mutagenesis, Insertional / methods* Plasmids / genetics Promoter Regions, Genetic Recombination, Genetic* Schizosaccharomyces / genetics* Schizosaccharomyces pombe Proteins / genetics Thiamine / metabolism Viral Proteins / genetics* Viral Proteins / metabolism
IF 1.255
Times Cited 35
WOS Category CHEMISTRY, APPLIED FOOD SCIENCE & TECHNOLOGY BIOTECHNOLOGY & APPLIED MICROBIOLOGY BIOCHEMISTRY & MOLECULAR BIOLOGY
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