Generation of temperature sensitive mutations with error-prone PCR in a gene encoding a component of the spindle pole body in fission yeast

Ngang Heok Tang; Chii Shyang Fong; Hirohisa Masuda; Isabelle Jourdain; Masashi Yukawa; Takashi Toda

doi:10.1080/09168451.2019.1611414

Generation of temperature sensitive mutations with error-prone PCR in a gene encoding a component of the spindle pole body in fission yeast

Biosci Biotechnol Biochem. 2019 Sep;83(9):1717-1720. doi: 10.1080/09168451.2019.1611414. Epub 2019 May 1.

Authors

Ngang Heok Tang¹, Chii Shyang Fong², Hirohisa Masuda³, Isabelle Jourdain⁴, Masashi Yukawa⁵, Takashi Toda⁵

Affiliations

¹ a Section of Neurobiology, Division of Biological Sciences, University of California , San Diego , CA , USA.
² b Cell Biology Program, Memorial Sloan Kettering Cancer Center , New York , NY , USA.
³ c Telomere Biology Section, Laboratory of Biochemistry and Molecular Biology, National Cancer Institute, National Institutes of Health , Bethesda , MD , USA.
⁴ d Biosciences, College of Life and Environmental Sciences, University of Exeter , Exeter , UK.
⁵ e Hiroshima Research Center for Healthy Aging (HiHA), Division of Biological and Life Sciences, Graduate School of Integrated Sciences for Life , Higashi-Hiroshima , Japan.

PMID: 31042107
DOI: 10.1080/09168451.2019.1611414

Abstract

Temperature-sensitive (ts) mutants provide powerful tools for investigation of cellular functions of essential genes. We report here asimple procedure to generate ts mutations using error-prone PCR within pcp1 that encodes aspindle pole body (SPB) component in Schizosaccharomyces pombe. This manipulation is not restricted to pcp1, and can be suited to any essential genes involved in other processes.

Keywords: Error-prone PCR; Pcp1; fission yeast; temperature sensitive mutants; the spindle pole body.

MeSH terms

Cell Cycle Proteins
Genes, Fungal*
Mutation*
Nuclear Proteins / genetics
Polymerase Chain Reaction / methods*
Schizosaccharomyces / genetics*
Schizosaccharomyces pombe Proteins / genetics
Spindle Pole Bodies / metabolism*
Temperature*

Substances

Cell Cycle Proteins
Nuclear Proteins
Schizosaccharomyces pombe Proteins
pcp1 protein, S pombe