A toolbox of stable integration vectors in the fission yeast Schizosaccharomyces pombe

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Version: Final published version
License: CC BY-NC 4.0
Serval ID
serval:BIB_AB39F7250738
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
A toolbox of stable integration vectors in the fission yeast Schizosaccharomyces pombe
Journal
Journal of Cell Science
Author(s)
Vještica Aleksandar, Marek Magdalena, Nkosi Pedro Junior, Merlini Laura, Liu Gaowen, Bérard Melvin, Billault-Chaumartin Ingrid, Martin Sophie G.
ISSN
0021-9533
1477-9137
ISSN-L
0021-9533
Publication state
Published
Issued date
01/01/2020
Peer-reviewed
Oui
Volume
133
Number
1
Pages
jcs240754
Language
english
Notes
Publication types: Journal Article
Publication Status: epublish
Abstract
Schizosaccharomyces pombe is a widely used model organism to study many aspects of eukaryotic cell physiology. Its popularity as an experimental system partially stems from the ease of genetic manipulations, where the innate homology-targeted repair is exploited to precisely edit the genome. While vectors to incorporate exogenous sequences into the chromosomes are available, most are poorly characterized. Here we show that commonly used fission yeast vectors, which upon integration produce repetitive genomic regions, yield unstable genomic loci. We overcome this problem by designing a new series of Stable Integration Vectors (SIV) that target four different prototrophy genes. SIV produce non-repetitive, stable genomic loci and integrate predominantly as single copy. Additionally, we develop a set of complementary auxotrophic alleles that preclude false-positive integration events. We expand the vector series to include antibiotic resistance markers, promoters, fluorescent tags and terminators, and build a highly modular toolbox to introduce heterologous sequences. Finally, as proof of concept, we generate a large set of ready-to-use, fluorescent probes to mark organelles and cellular processes with a wide range of applications in fission yeast research.
Keywords
Cell Biology
Pubmed
Web of science
Funding(s)
Swiss National Science Foundation / 310030B_176396
Create date
15/12/2019 17:56
Last modification date
08/02/2020 6:17
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