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

Abstract

Transformation (i.e., genetic modification of a cell by the incorporation of exogenous DNA) is indispensable for manipulating fungi. Here, we review the transformation methods for Saccharomyces cerevisiae, Schizosaccharomyces pombe, Candida albicans, Pichia pastoris and Aspergillus species and discuss some common modifications to improve transformation efficiency. We also present a model of the mechanism underlying S. cerevisiae transformation, based on recent reports and the mechanism of transfection in mammalian systems. This model predicts that DNA attaches to the cell wall and enters the cell via endocytotic membrane invagination, although how DNA reaches the nucleus is unknown. Polyethylene glycol is indispensable for successful transformation of intact cells and the attachment of DNA and also possibly acts on the membrane to increase the transformation efficiency. Both lithium acetate and heat shock, which enhance the transformation efficiency of intact cells but not that of spheroplasts, probably help DNA to pass through the cell wall.

• Authors: Shigeyuki Kawai, Wataru Hashimoto, Kousaku Murata
• Journal: Bioengineered bugs (Bioeng Bugs) Vol. 1 Issue 6 Pg. 395-403 ( 2010) ISSN: 1949-1026 [Electronic] United States
• PMID: 21468206 (Publication Type: Journal Article, Review)
• Copyright: © 2010 Landes Bioscience

Chemical References

• Acetates
• DNA, Fungal
• Polyethylene Glycols
• lithium acetate

Topics

• Acetates (metabolism)
• Cell Wall (metabolism)
• DNA, Fungal (genetics)
• Electroporation
• Endocytosis
• Fungi (classification, genetics, metabolism)
• Polyethylene Glycols (metabolism)
• Saccharomyces cerevisiae (genetics, metabolism)
• Spheroplasts (genetics, metabolism)
• Transformation, Genetic