Abstract
There are several ways to introduce non-native DNA into yeast cells, including chemical transformation and electroporation. Methods for both of these procedures are outlined in this chapter. Both methods permit the uptake of DNA from the environment through yeast cell membranes and this DNA can be episomally maintained or integrated into the host genome. However, yeast cells must first be made competent to permit passive entry of the DNA and various methods are outlined in this chapter to facilitate this. All of the described methods can be applied in combination with antibiotic or auxotrophic selection pressure.
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Baron M, Reynes JP, Stassi D, Tiraby G (1992) A selectable bifunctional beta-galactosidase::phleomycin-resistance fusion protein as a potential marker for eukaryotic cells. Gene 114:239–243
Drocourt D, Calmels T, Reynes JP, Baron M, Tiraby G (1990) Cassettes of the Streptoalloteichus hindustanus ble gene for transformation of lower and higher eukaryotes to phleomycin resistance. Nucleic Acids Res 18:4009
Mulsant P, Gatignol A, Dalens M, Tiraby G (1988) Phleomycin resistance as a dominant selectable marker in CHO cells. Somat Cell Mol Genet 14:243–252
Perez P, Tiraby G, Kallerhoff J, Perret J (1989) Phleomycin resistance as a dominant selectable marker for plant-cell transformation. Plant Mol Biol 13:365–373
Lin-Cereghino J, Wong WW, Xiong S, Giang W, Luong LT, Vu J, Johnson SD, Lin-Cereghino GP (2005) Condensed protocol for competent cell preparation and transformation of the methylotrophic yeast Pichia pastoris. Biotechniques 38:44, 46, 48
Weiss HM, Haase W, Michel H, Reilander H (1998) Comparative biochemical and pharmacological characterization of the mouse 5HT(5A) 5-hydroxytryptamine receptor and the human beta(2)-adrenergic receptor produced in the methylotrophic yeast Pichia pastoris. Biochem J 330:1137–1147
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Jamshad, M., Darby, R.A.J. (2012). Yeast Transformation to Generate High-Yielding Clones. In: Bill, R. (eds) Recombinant Protein Production in Yeast. Methods in Molecular Biology, vol 866. Humana Press. https://doi.org/10.1007/978-1-61779-770-5_6
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DOI: https://doi.org/10.1007/978-1-61779-770-5_6
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Online ISBN: 978-1-61779-770-5
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