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A toolbox of Stable Integration Vectors (SIV) in the fission yeast Schizosaccharomyces pombe

View ORCID ProfileAleksandar Vještica, View ORCID ProfileMagdalena Marek, Pedro N’kosi, View ORCID ProfileLaura Merlini, View ORCID ProfileGaowen Liu, Melvin Bérard, Ingrid Billault-Chaumartin, View ORCID ProfileSophie G Martin
doi: https://doi.org/10.1101/808329
Aleksandar Vještica
Department of Fundamental Microbiology, University of Lausanne, Biophore building, CH-1015 Lausanne, Switzerland
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  • For correspondence: aleksandar.vjestica@unil.ch sophie.martin@unil.ch
Magdalena Marek
Department of Fundamental Microbiology, University of Lausanne, Biophore building, CH-1015 Lausanne, Switzerland
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  • ORCID record for Magdalena Marek
Pedro N’kosi
Department of Fundamental Microbiology, University of Lausanne, Biophore building, CH-1015 Lausanne, Switzerland
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Laura Merlini
Department of Fundamental Microbiology, University of Lausanne, Biophore building, CH-1015 Lausanne, Switzerland
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Gaowen Liu
Department of Fundamental Microbiology, University of Lausanne, Biophore building, CH-1015 Lausanne, Switzerland
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Melvin Bérard
Department of Fundamental Microbiology, University of Lausanne, Biophore building, CH-1015 Lausanne, Switzerland
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Ingrid Billault-Chaumartin
Department of Fundamental Microbiology, University of Lausanne, Biophore building, CH-1015 Lausanne, Switzerland
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Sophie G Martin
Department of Fundamental Microbiology, University of Lausanne, Biophore building, CH-1015 Lausanne, Switzerland
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  • ORCID record for Sophie G Martin
  • For correspondence: aleksandar.vjestica@unil.ch sophie.martin@unil.ch
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Abstract

Schizosaccharomyces pombe is a widely used model organism that resembles higher eukaryotes in many aspects of 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.

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Posted October 18, 2019.
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A toolbox of Stable Integration Vectors (SIV) in the fission yeast Schizosaccharomyces pombe
Aleksandar Vještica, Magdalena Marek, Pedro N’kosi, Laura Merlini, Gaowen Liu, Melvin Bérard, Ingrid Billault-Chaumartin, Sophie G Martin
bioRxiv 808329; doi: https://doi.org/10.1101/808329
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A toolbox of Stable Integration Vectors (SIV) in the fission yeast Schizosaccharomyces pombe
Aleksandar Vještica, Magdalena Marek, Pedro N’kosi, Laura Merlini, Gaowen Liu, Melvin Bérard, Ingrid Billault-Chaumartin, Sophie G Martin
bioRxiv 808329; doi: https://doi.org/10.1101/808329

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