Abstract
We have compared a number of procedures for the transformation of whole cells of the yeast Saccharomyces cerevisiae and assessed the effects of dimethylsulphoxide (DMSO) or ethanol, both of which have been reported to enhance transformation efficiency. We find that simplified methods benefit from the addition of one of these compounds, and although differences are observed between strains as to the more beneficial reagent, peak transformation efficiency is, in general obtained with 10% DMSO or 10% EtOH. Increases of between six- and 50-fold are observed, despite a reduction in cell viability, and at this concentration the two compounds are not additive in their effects. The optimum level appears to depend on a balance between improved DNA uptake and reduced cell viability. As a result of this work we present a straightforward and rapid transformation procedure.
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References
Altherr MR, Quinn LA, Kado CI, Rodroguez RL (1983) NATO Adv Sci Inst Series A 61:33–36
Baldari C, Murray JAH, Ghiara P, Cesareni G, Galeotti CL (1987) EMBO J 6:229–234
Beggs JD (1978) Nature 275:104–109
Brzobohaty B, Kovac L (1986) J Gen Microbiol 132:3089–3093
Chen D-C, Yang B-C, Kuo T-T (1992) Curr Genet 21:83–84
Chung CT, Miller RH (1988) Nucleic Acids Res 16:3580
De Nobel JG, Dijkers C, Hooijberg E, Klis FM (1989) J Gen Microbiol 135:2077–2084
Delorme E (1989) Appl Environ Microbiol 55:2242–2246
Elble R (1992) Bio Techniques 13:18–20
Gietz RD, Schiestl RH (1991) Yeast 7:253–263
Gietz RD, St. Jean A, Woods RA, Schiestl RH (1992) Nucleic Acids Res 20:1425
Hasimoto H, Morikawa H, Yamada Y, Kimura A (1985) Appl Microbiol Biotechnol 21:336–339
Hill DE (1989) Nucleic Acids Res 17:8011
Hill J, Jan KA, Donald G, Griffiths DE (1991) Nucleic Acids Res 19:5791
Hinnen A, Hicks JB, Fink GR (1978) Proc Natl Acad Sci USA 75:1929–1933
Ingram LO, Buttke TM (1984) Adv Microb Physiol 25:253–300
Ito H, Fukuda Y, Murata K, Kimura A (1983a) J Bacteriol 153:163–168
Ito H, Murata K, Kimura A (1983b) Agric Biol Chem 47:1691–1692
Ito H, Murata K, Kimura A (1984) Agric Biol Chem 48:341–347
Jones RP, Greenfield PF (1987) Yeast 3:223–232
Keszenman-Pereyra D, Hieda K (1988) Curr Genet 13:21–23
Keszenman-Pereyra D, Hieda K (1989) Agric Biol Chem 53:2535–2536
Lauermann V (1991) Curr Genet 20:1–3
Lopata MA, Cleveland DW, Sollner-Webb B (1984) Nucleic Acids Res 12:5707–5717
Meilhoc E, Masson J-M (1990) Biotechnology 8:223–227
Minet M, Dufour M-E, Lacroute F (1992) Plant J 2:417–422
Murray JAH, Scarpa M, Rossi N, Cesareni G (1987) EMBO J 6:4205–4212
Ogas J, Andrews BJ, Herskowitz I (1991) Cell 66:1015–1026
Schiestl RH, Gietz RD (1989) Curr Genet 16:339–346
Xiong Y, Connolly T, Futcher B, Beach D (1991) Cell 65:691–699
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Communicated by A. Hinnen
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Soni, R., Carmichael, J.P. & Murray, J.A.H. Parameters affecting lithium acetate-mediated transformation of Saccharomyces cerevisiae and development of a rapid and simplified procedure. Curr Genet 24, 455–459 (1993). https://doi.org/10.1007/BF00351857
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DOI: https://doi.org/10.1007/BF00351857