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An improved tetracycline-inducible expression system for fission yeast.

Authors :
Lyu XH
Yang YS
Pan ZQ
Ning SK
Suo F
Du LL
Source :
Journal of cell science [J Cell Sci] 2024 Nov 01; Vol. 137 (21). Date of Electronic Publication: 2024 Nov 01.
Publication Year :
2024

Abstract

The ability to manipulate gene expression is valuable for elucidating gene function. In the fission yeast Schizosaccharomyces pombe, the most widely used regulatable expression system is the nmt1 promoter and its two attenuated variants. However, these promoters have limitations, including a long lag, incompatibility with rich media and unsuitability for non-dividing cells. Here, we present a tetracycline-inducible system free of these shortcomings. Our system features the enotetS promoter, which achieves a similar induced level and a higher induction ratio compared to the nmt1 promoter, without exhibiting a lag. Additionally, our system includes four weakened enotetS variants, offering an expression range similar to that of the nmt1 series promoters but with more intermediate levels. To enhance usability, each promoter is combined with a Tet-repressor-expressing cassette in an integration plasmid. Importantly, our system can be used in non-dividing cells, enabling the development of a synchronous meiosis induction method with high spore viability. Moreover, our system allows for the shutdown of gene expression and the generation of conditional loss-of-function mutants. This system provides a versatile and powerful tool for manipulating gene expression in fission yeast.<br />Competing Interests: Competing interests The authors declare no competing or financial interests.<br /> (© 2024. Published by The Company of Biologists Ltd.)

Details

Language :
English
ISSN :
1477-9137
Volume :
137
Issue :
21
Database :
MEDLINE
Journal :
Journal of cell science
Publication Type :
Academic Journal
Accession number :
39318285
Full Text :
https://doi.org/10.1242/jcs.263404