Literature DB >> 16233287

Extremely simple, rapid and highly efficient transformation method for the yeast Saccharomyces cerevisiae using glutathione and early log phase cells.

Yoshiyuki Hayama1, Yasuki Fukuda, Shigeyuki Kawai, Wataru Hashimoto, Kousaku Murata.   

Abstract

In the presence of polyethylene glycol (PEG), budding cells of Saccharomyces cerevisiae in the early log phase were transformed by exogenous plasmid DNA without additional specific chemical or physical treatments. This capacity of the yeast cells to become competent was strictly dependent on the growth phase, being induced in the early log phase, becoming maximum between the early and mid log phases and then disappearing rapidly in the mid log phase. The transformation was most efficient at pH 6 and the frequency increased with increasing DNA and cell concentrations. PEGs with average molecular sizes between 1000 and 3500 showed almost the same effects and were used most efficiently at 35%. The transformation frequency of S. cerevisiae was markedly enhanced when the oxidized form of glutathione (GSSG), but not the reduced form, was included in the mixture comprising early log phase cells, plasmid DNA, and PEG, and the transformation system with GSSG could be used as a convenient transformation method for the yeast S. cerevisiae.

Entities:  

Year:  2002        PMID: 16233287     DOI: 10.1263/jbb.94.166

Source DB:  PubMed          Journal:  J Biosci Bioeng        ISSN: 1347-4421            Impact factor:   2.894


  6 in total

1.  The role of cell wall revealed by the visualization of Saccharomyces cerevisiae transformation.

Authors:  Tuan Anh Pham; Shigeyuki Kawai; Emi Kono; Kousaku Murata
Journal:  Curr Microbiol       Date:  2010-11-16       Impact factor: 2.188

2.  Visualization of the synergistic effect of lithium acetate and single-stranded carrier DNA on Saccharomyces cerevisiae transformation.

Authors:  Tuan Anh Pham; Shigeyuki Kawai; Kousaku Murata
Journal:  Curr Genet       Date:  2011-05-12       Impact factor: 3.886

Review 3.  Ecologically driven competence for exogenous DNA uptake in yeast.

Authors:  Petar Tomev Mitrikeski
Journal:  Curr Microbiol       Date:  2015-04-01       Impact factor: 2.188

Review 4.  Transformation of Saccharomyces cerevisiae and other fungi: methods and possible underlying mechanism.

Authors:  Shigeyuki Kawai; Wataru Hashimoto; Kousaku Murata
Journal:  Bioeng Bugs       Date:  2010 Nov-Dec

5.  Enhancement of plasmid DNA transformation efficiencies in early stationary-phase yeast cell cultures.

Authors:  Jennifer DeMars Tripp; Jennifer L Lilley; Whitney N Wood; L Kevin Lewis
Journal:  Yeast       Date:  2013-04-12       Impact factor: 3.239

6.  CRISPR/Cas9 System as a Valuable Genome Editing Tool for Wine Yeasts with Application to Decrease Urea Production.

Authors:  Ileana Vigentini; Marinella Gebbia; Alessandra Belotti; Roberto Foschino; Frederick P Roth
Journal:  Front Microbiol       Date:  2017-11-09       Impact factor: 5.640

  6 in total

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