DOI: 10.1002/yea.1233. New drug‐resistant cassettes for gene disruption and epitope tagging in <i>Schizosaccharomyces pombe</i>

New drug‐resistant cassettes for gene disruption and epitope tagging in Schizosaccharomyces pombe

10.1002/yea.1233

Crossref journal-article

Wiley

Yeast (311)

Abstract

AbstractWe describe new heterologous modules for PCR‐based gene targeting in the fission yeast Schizosaccharomyces pombe. Two bacterial genes, hph and nat, which display dominant drug‐resistance phenotypes, are used as new selectable markers in these modules. Both genes have been used successfully in the budding yeast Saccharomyces cerevisiae, in which hph confers resistance to hygromycin B, while nat confers nourseothricin resistance (Goldstein and McCusker, 1999). Vector modules for gene disruption and C‐terminal tagging with 3HA, 13Myc and GFP(S65T) are constructed using previously constructed pFA6a–MX6–derived plasmids (Bähler et al., 1998; Wach et al., 1997). In combination with the existing systems that are based upon the G418‐resistance gene (kan), triple gene deletions or tags could be constructed. In addition a vector for one‐step integration of a monomeric RFP (mRFP) to the C‐terminus of proteins of interest is developed. Finally, oligonucleotides that allow a simple marker switch from kan to hph or nat, and vice versa, are described. The new constructs developed here should facilitate post‐genomic molecular analysis of protein functions in fission yeast. Copyright © 2005 John Wiley & Sons, Ltd.

Bibliography

Sato, M., Dhut, S., & Toda, T. (2005). New drugâresistant cassettes for gene disruption and epitope tagging in Schizosaccharomyces pombe. Yeast, 22(7), 583â591. Portico.

Authors 3

Masamitsu Sato (first)
Susheela Dhut (additional)
Takashi Toda (additional)

References 27 Referenced 242

10.1073/pnas.97.22.11984
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10.1073/pnas.082243699
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10.1093/genetics/136.3.849
10.1002/yea.983
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{'key': 'e_1_2_6_15_1', 'first-page': '10 857', 'article-title': 'nmt1 of fission yeast', 'volume': '265', 'author': 'Maundrell K', 'year': '1990', 'journal-title': 'J Biol Chem'} / J Biol Chem / nmt1 of fission yeast by Maundrell K (1990)
{'key': 'e_1_2_6_16_1', 'first-page': '773', 'article-title': 'Molecular genetic analyses of fission yeast Schizosaccharomyces pombe', 'volume': '194', 'author': 'Moreno S', 'year': '1991', 'journal-title': 'Methods Enzymol'} / Methods Enzymol / Molecular genetic analyses of fission yeast Schizosaccharomyces pombe by Moreno S (1991)
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{'key': 'e_1_2_6_21_1', 'volume-title': 'Molecular Cloning: A Laboratory Manual', 'author': 'Sambrook J', 'year': '1989'} / Molecular Cloning: A Laboratory Manual by Sambrook J (1989)
10.1016/S0092-8674(00)80320-7
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10.1016/0092-8674(83)90331-8
10.1002/(SICI)1097-0061(19970915)13:11<1065::AID-YEA159>3.0.CO;2-K
10.1002/yea.320101310
10.1073/pnas.1534703100
10.1091/mbc.E04-10-0859

Dates

Type	When
Created	20 years, 2 months ago (June 7, 2005, 2:17 p.m.)
Deposited	1 year, 10 months ago (Oct. 8, 2023, 12:41 a.m.)
Indexed	3 weeks, 1 day ago (Aug. 2, 2025, 12:58 a.m.)
Issued	20 years, 3 months ago (May 1, 2005)
Published	20 years, 3 months ago (May 1, 2005)
Published Online	20 years, 2 months ago (June 7, 2005)
Published Print	20 years, 3 months ago (May 1, 2005)

Funders 0

None

BibTeX

@article{Sato_2005, title={New drug‐resistant cassettes for gene disruption and epitope tagging in Schizosaccharomyces pombe}, volume={22}, ISSN={1097-0061}, url={http://dx.doi.org/10.1002/yea.1233}, DOI={10.1002/yea.1233}, number={7}, journal={Yeast}, publisher={Wiley}, author={Sato, Masamitsu and Dhut, Susheela and Toda, Takashi}, year={2005}, month=may, pages={583–591} }

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