Abstract
Since it was initially established, the lithium method for transforming Saccharomyces cerevisiae intact cells has been modified, and a possible underlying mechanism has been elucidated. This method requires polyethylene glycol (PEG) and transformation efficiency is enhanced by lithium and single-stranded carrier DNA (ssDNA). Here, we describe the original lithium method, the modified method, and the possible underlying mechanism, in which plasmid DNA that was absorbed onto the cell wall enters into the cells via endocytotic membrane invagination and lithium and ssDNA synergistically alter cell wall structure and enhance transformation efficiency.
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References
Gietz RD, Woods RA (2002) Transformation of yeast by lithium acetate/single-stranded carrier DNA/polyethylene glycol method. Methods Enzymol 350:87–96
Gietz D, St Jean A, Woods RA, Schiestl RH (1992) Improved method for high efficiency transformation of intact yeast cells. Nucleic Acids Res 20:1425
Gietz RD, Schiestl RH, Willems AR, Woods RA (1995) Studies on the transformation of intact yeast cells by the LiAc/SS-DNA/PEG procedure. Yeast 11:355–360
Gurrieri S, Wells KS, Johnson ID, Bustamante C (1997) Direct visualization of individual DNA molecules by fluorescence microscopy: characterization of the factors affecting signal/background and optimization of imaging conditions using YOYO. Anal Biochem 249:44–53
Hayama Y, Fukuda Y, Kawai S, Hashimoto W, Murata K (2002) Extremely simple, rapid and highly efficient transformation method for the yeast Saccharomyces cerevisiae using glutathione and early log phase cells. J Biosci Bioeng 94:166–171
Hinnen A, Hicks JB, Fink GR (1978) Transformation of yeast. Proc Natl Acad Sci U S A 75:1929–1933
Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168
Kawai S, Pham TA, Nguyen HT, Nankai H, Utsumi T, Fukuda Y, Murata K (2004) Molecular insights on DNA delivery into Saccharomyces cerevisiae. Biochem Biophys Res Commun 317:100–107
Kawai S, Hashimoto W, Murata K (2010) Transformation of Saccharomyces cerevisiae and other fungi: methods and possible underlying mechanism. Bioeng Bugs 1:395–403
Pham TA, Kawai S, Kono E, Murata K (2011a) The role of cell wall revealed by the visualization of Saccharomyces cerevisiae transformation. Curr Microbiol 62:956–961
Pham TA, Kawai S, Murata K (2011b) Visualization of the synergistic effect of lithium acetate and single-stranded carrier DNA on Saccharomyces cerevisiae transformation. Curr Genet 57:233–239
Schiestl RH, Gietz RD (1989) High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier. Curr Genet 16:339–346
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Kawai, S., Murata, K. (2015). Transformation of Intact Cells of Saccharomyces cerevisiae: Lithium Methods and Possible Underlying Mechanism. In: van den Berg, M., Maruthachalam, K. (eds) Genetic Transformation Systems in Fungi, Volume 1. Fungal Biology. Springer, Cham. https://doi.org/10.1007/978-3-319-10142-2_18
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DOI: https://doi.org/10.1007/978-3-319-10142-2_18
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Online ISBN: 978-3-319-10142-2
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