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Transformation of intact yeast cells treated with alkali cations
Open accessAbstract
Intact yeast cells treated with alkali cations took up plasmid DNA. Li+, Cs+, Rb+, K+, and Na+ were effective in inducing competence. Conditions for the transformation of Saccharomyces cerevisiae D13-1A with plasmid YRp7 were studied in detail with CsCl. The optimum incubation time was 1 h, and the optimum cell concentration was 5 x 10(7) cells per ml. The optimum concentration of Cs+ was 1.0 M. Transformation efficiency increased with increasing concentrations of plasmid DNA. Polyethylene glycol was absolutely required. Heat pulse and various polyamines or basic proteins stimulated the uptake of plasmid DNA. Besides circular DNA, linear plasmid DNA was also taken up by Cs+-treated yeast cells, although the uptake efficiency was considerably reduced. The transformation efficiency with Cs+ or Li+ was comparable with that of conventional protoplast methods for a plasmid containing ars1, although not for plasmids containing a 2 microns origin replication.
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Cited by
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Uetz, P., Giot, L., Cagney, G., Mansfield, T. A., Judson, R. S., Knight, J. R., Lockshon, D., Narayan, V., Srinivasan, M., Pochart, P., et al. (2000) · Nature · doi:10.1038/35001009 · Cited by 4,706
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Gietz, R. D., & Schiestl, R. H. (2007) · Nature Protocols · doi:10.1038/nprot.2007.13 · Cited by 2,772
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Hill, J. E., Myers, A. M., Koerner, T. J., & Tzagoloff, A. (1986) · Yeast · doi:10.1002/yea.320020304 · Cited by 1,382
Bishop, D. K., Park, D., Xu, L., & Kleckner, N. (1992) · Cell · doi:10.1016/0092-8674(92)90446-j · Cited by 1,144
Alani, E., Cao, L., & Kleckner, N. (1987) · Genetics · doi:10.1534/genetics.112.541.test · Cited by 873
Metadata
- Authors
- Ito, H., Fukuda, Y., Murata, K., & Kimura, A.
- Venue
- Journal of Bacteriology
- Publisher
- American Society for Microbiology
- Published
- 1983-01
- Volume
- 153
- Issue
- 1
- Pages
- 163-168
- ISSN
- 0021-9193 (ISSN-L)
- 1098-5530
- OpenAlex
- W1973782344
- Cited by
- 7,518
- References
- 0
- FWCI
- 54.58
- Type
- journal article
- Language
- en
- Open access
- bronze
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- Ito, H., Fukuda, Y., Murata, K., & Kimura, A. (1983). Transformation of intact yeast cells treated with alkali cations. Journal of Bacteriology. https://doi.org/10.1128/jb.153.1.163-168.1983
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