title: 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.affiliation: Laboratory of Basic and Applied Molecular Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
authors.forename: Shigeyuki
authors.initials: S
authors.lastname: Kawai
authors.forename: Wataru
authors.initials: W
authors.lastname: Hashimoto
authors.forename: Kousaku
authors.initials: K
authors.lastname: Murata
mesh_terms: D000085:Acetates / Q000378:metabolism; D002473:Cell Wall / Q000378:metabolism; D004271:DNA, Fungal / Q000235:genetics; D018274:Electroporation; D004705:Endocytosis; D005658:Fungi / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D011092:Polyethylene Glycols / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D013104:Spheroplasts / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic
chemical_list: D000085:Acetates; D004271:DNA, Fungal; D011092:Polyethylene Glycols; C488804:lithium acetate