Transformation ofSchizosaccharomyces pombewith DNA requires the conditioning of cells to promote DNA uptake followed by cell growth under conditions that select and maintain the plasmid or integration event. The three main methodologies are electroporation, treatment with lithium cations, and transformation of protoplasts. The lithium acetate method described here is widely used because it is simple and reliable.
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http://dx.doi.org/10.1101/pdb.prot090969 | DOI Listing |
Methods Mol Biol
November 2024
Laboratory of Molecular and Chemical Cell Biology, Graduate School of Integrated Sciences for Life, Hiroshima University, Hiroshima, Japan.
Kinesin-5 motor proteins are essential for mitotic spindle formation and maintenance, ensuring accurate chromosome segregation. Human kinesin-5 is highly expressed in various cancer cells but not in nonproliferative tissues; therefore, it is expected to be an attractive target for cancer chemotherapy, with fewer adverse side effects. Many inhibitors have been developed and subjected to clinical trials; however, they have not yet been commercially distributed because of their poor efficacy and frequent drug resistance.
View Article and Find Full Text PDFMethods Mol Biol
November 2024
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, USA.
We present a protocol to perform CRISPR/Cas9-mediated genome editing in the fission yeast Schizosaccharomyces pombe that does not require cloning and uses the fluoride exporter channel Fex1 as the selection marker. Transformation is typically carried out on the same day of PCR primer arrival and successfully edited strains are selected 5 days after transformation. We expect the adoption of this protocol to further accelerate the throughput of genome editing in S.
View Article and Find Full Text PDFMethods Mol Biol
November 2024
Molecular Cellular and Developmental Biology Program, Division of Biology, Kansas State University, Manhattan, KS, USA.
Proc Natl Acad Sci U S A
November 2024
National Institute of Biological Sciences, Beijing 102206, China.
Biomolecules
August 2024
Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, Little Rock, AR 72205-7199, USA.
Recombination hotspot-activating DNA sites (e.g., , , ) and their binding proteins (e.
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