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

Fingerprint

Schizosaccharomyces
Schizosaccharomyces pombe
gene targeting
Genes
promoter regions
genetic markers
genes
isozymes
chromosomes
Functional analysis
genome
Genetic Recombination
Isoenzymes
Chromosomes
Genome
Cre recombinase

All Science Journal Classification (ASJC) codes

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

Cite this

A set of loxP marker cassettes for cre-mediated multiple gene disruption in Schizosaccharomyces pombe. / Iwaki, Tomoko; Takegawa, Kaoru.

In: Bioscience, Biotechnology and Biochemistry, Vol. 68, No. 3, 01.03.2004, p. 545-550.

Research output: Contribution to journalArticle

@article{d271a15171894ad39cfb7a7ec32fc6f5,
title = "A set of loxP marker cassettes for cre-mediated multiple gene disruption in Schizosaccharomyces pombe",
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.",
author = "Tomoko Iwaki and Kaoru Takegawa",
year = "2004",
month = "3",
day = "1",
doi = "10.1271/bbb.68.545",
language = "English",
volume = "68",
pages = "545--550",
journal = "Bioscience, Biotechnology and Biochemistry",
issn = "0916-8451",
publisher = "Japan Society for Bioscience Biotechnology and Agrochemistry",
number = "3",

}

TY - JOUR

T1 - A set of loxP marker cassettes for cre-mediated multiple gene disruption in Schizosaccharomyces pombe

AU - Iwaki, Tomoko

AU - Takegawa, Kaoru

PY - 2004/3/1

Y1 - 2004/3/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=4544273971&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=4544273971&partnerID=8YFLogxK

U2 - 10.1271/bbb.68.545

DO - 10.1271/bbb.68.545

M3 - Article

VL - 68

SP - 545

EP - 550

JO - Bioscience, Biotechnology and Biochemistry

JF - Bioscience, Biotechnology and Biochemistry

SN - 0916-8451

IS - 3

ER -