Review

Transformation of Saccharomyces cerevisiae and other fungi: Methods and possible underlying mechanism

Volume 1, Issue 6   November/December 2010
Pages 395 - 403
http://dx.doi.org/10.4161/bbug.1.6.13257
Authors: Shigeyuki Kawai, Wataru Hashimoto and Kousaku Murata

View affiliations

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.

Received: September 30, 2010; Accepted: August 5, 2010

Preview:



Comments

Please Log In to comment on this article.
If you do not have an account Create One Here.
blog comments powered by Disqus
Advertisements