酿酒酵母及其他真菌的转化：方法及可能的潜在机制
Transformation of Saccharomyces cerevisiae and other fungi: methods and possible underlying mechanism.
作者信息
Kawai Shigeyuki, Hashimoto Wataru, Murata Kousaku
机构信息
Laboratory of Basic and Applied Molecular Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
出版信息
Bioeng Bugs. 2010 Nov-Dec;1(6):395-403. doi: 10.4161/bbug.1.6.13257.
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.
转化（即通过掺入外源DNA对细胞进行基因改造）对于操纵真菌而言必不可少。在此，我们综述了酿酒酵母、粟酒裂殖酵母、白色念珠菌、巴斯德毕赤酵母和曲霉属的转化方法，并讨论了一些用于提高转化效率的常见改良方法。我们还基于近期报道及哺乳动物系统中的转染机制，提出了酿酒酵母转化潜在机制的模型。该模型预测DNA附着于细胞壁并通过内吞性膜内陷进入细胞，尽管DNA如何到达细胞核尚不清楚。聚乙二醇对于完整细胞的成功转化以及DNA的附着必不可少，并且可能还作用于细胞膜以提高转化效率。醋酸锂和热休克均可提高完整细胞的转化效率，但对原生质体无效，它们可能有助于DNA穿过细胞壁。
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