Literature DB >> 29423856

High-Frequency Lithium Acetate Transformation of Schizosaccharomyces pombe.

Sudhir Kumar Rai1,2, Angela Atwood-Moore1, Henry L Levin3.   

Abstract

The introduction of ectopic DNA, such as plasmids, into yeast cells has for decades been a critical protocol for the study of this eukaryotic model system. We describe here an efficient transformation procedure for use in the fission yeast Schizosaccharomyces pombe. This method relies on chemical agents (lithium acetate, and polyethylene glycol) and temperature stresses, which ultimately facilitate transfer of the genetic material through the cell wall and plasma membrane without significant impact on the transferred DNA or the recipient cell. Using this protocol, we consistently see transformation efficiencies between 1.0 × 103 and 1.0 × 104 transformants per microgram of the plasmid with 108 S. pombe cells. The principal benefits and advantages of this method are its simplicity, efficiency, and relative speed of completion.

Entities:  

Keywords:  Fission yeast; Lithium acetate; Polyethylene glycol; Schizosaccharomyces pombe; Transformation; pFL20

Mesh:

Substances:

Year:  2018        PMID: 29423856      PMCID: PMC6388634          DOI: 10.1007/978-1-4939-7546-4_15

Source DB:  PubMed          Journal:  Methods Mol Biol        ISSN: 1064-3745


  4 in total

1.  The PPIP5K Family Member Asp1 Controls Inorganic Polyphosphate Metabolism in S. pombe.

Authors:  Marina Pascual-Ortiz; Eva Walla; Ursula Fleig; Adolfo Saiardi
Journal:  J Fungi (Basel)       Date:  2021-07-31

2.  Fission Yeast Schizosaccharomyces pombe: A Unicellular "Micromammal" Model Organism.

Authors:  Aditi Vyas; Anna V Freitas; Zachary A Ralston; Zhaohua Tang
Journal:  Curr Protoc       Date:  2021-06

3.  A protocol for transposon insertion sequencing in Schizosaccharomyces pombe to identify factors that maintain heterochromatin.

Authors:  Feng Li; Stevephen Hung; Caroline Esnault; Henry L Levin
Journal:  STAR Protoc       Date:  2021-03-23

4.  Dense Transposon Integration Reveals Essential Cleavage and Polyadenylation Factors Promote Heterochromatin Formation.

Authors:  Si Young Lee; Stevephen Hung; Caroline Esnault; Rakesh Pathak; Kory R Johnson; Oluwadamilola Bankole; Akira Yamashita; Hongen Zhang; Henry L Levin
Journal:  Cell Rep       Date:  2020-02-25       Impact factor: 9.995

  4 in total

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