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Generation and characterisation of temperature sensitive mutants of genes encoding the fission yeast spindle pole body

, , , , ,
1 Section of Neurobiology, Division of Biological Sciences, University of California, San Diego, San Diego, CA, United States
2 Cell Biology Program, Memorial Sloan Kettering Cancer Center, New York, NY, United States
3 Telomere Biology Section, Laboratory of Biochemistry and Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD, United States
4 Biosciences, College of Life and Environmental Sciences, University of Exeter, Exeter, United Kingdom
5 Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, Hiroshima, Japan
DOI
10.7287/peerj.preprints.3377v1
Published
Accepted
Subject Areas
Cell Biology, Genetics
Keywords
fission yeast, PCR, temperature sensitive mutants, drug resistance, replica plating, spindle pole body, γ-tubulin complex
Copyright
© 2017 Tang et al.
Licence
This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ Preprints) and either DOI or URL of the article must be cited.
Cite this article
Tang NH, Fong CS, Masuda H, Jourdain I, Yukawa M, Toda T. 2017. Generation and characterisation of temperature sensitive mutants of genes encoding the fission yeast spindle pole body. PeerJ Preprints 5:e3377v1

Abstract

The spindle pole body (SPB) in fungi is the equivalent of the animal centrosome. A number of previous studies have identified many, if not all, components of the SPB. The SPB is the structural platform for microtubule nucleation and plays important roles, both in mitosis and meiosis. The SPB is absolutely essential for cell survival and its abnormalities give rise to aberrant cell division and morphogenesis. Therefore, it is crucial to understand how the SPB organises itself and how the functions of individual SPB components are regulated. We report here a procedure to generate temperature sensitive mutants in the fission yeast, Schizosaccharomyces pombe. The approach has proved useful to characterise functions of individual SPB components. This original genetic manipulation is however not restricted to analysis of SPB functions, and can be suited to investigate other cellular processes in S. pombe.

Author Comment

This manuscript describes genetic analysis of the fission yeast spindle pole body.This is a preprint submission to PeerJ Preprints.

Additional Information

Competing Interests

The authors declare that they have no competing interests.

Author Contributions

Ngang Heok Tang conceived and designed the experiments, performed the experiments, analyzed the data, contributed reagents/materials/analysis tools, wrote the paper, prepared figures and/or tables, reviewed drafts of the paper.

Chii Shyang Fong conceived and designed the experiments, performed the experiments, analyzed the data, contributed reagents/materials/analysis tools, wrote the paper, prepared figures and/or tables, reviewed drafts of the paper.

Hirohisa Masuda performed the experiments, contributed reagents/materials/analysis tools, reviewed drafts of the paper.

Isabelle Jourdain conceived and designed the experiments, performed the experiments, contributed reagents/materials/analysis tools, reviewed drafts of the paper.

Masashi Yukawa performed the experiments, contributed reagents/materials/analysis tools, reviewed drafts of the paper.

Takashi Toda conceived and designed the experiments, contributed reagents/materials/analysis tools, wrote the paper, reviewed drafts of the paper.

Data Deposition

The following information was supplied regarding data availability:

The raw data/code is included in the manuscript.

Funding

This work was supported by Cancer Research UK/the Francis Crick Institute, the Japan Society for the Promotion of Science KAKENHI Scientific Research (A) (16H02503), Challenging Exploratory Research (16K14672) (T.T.) and KAKENHI Scientific Research (C) (16K07694) (M.Y.). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Generation and characterisation of temperature sensitive mutants of genes encoding the fission yeast spindle pole body
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