Transformation of Saccharomyces Cerevisiae and Other Fungi: Methods and Possible Underlying Mechanism

Overview

Journal Bioeng Bugs

Specialties Biomedical Engineering
Biotechnology
Microbiology

Date 2011 Apr 7

PMID 21468206

Citations 83

Authors

Shigeyuki Kawai

Wataru Hashimoto

Kousaku Murata

Affiliations

Soon will be listed here.

Abstract

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.

Citing Articles

Mining yeast diversity unveils novel targets for improved heterologous laccase production in Saccharomyces cerevisiae.

Wong R, Foo M, Lay J, Wai T, Moore J, Dutreux F Microb Cell Fact. 2025; 24(1):60.

PMID: 40059166 PMC: 11892151. DOI: 10.1186/s12934-025-02677-1.

Protein targeting to Starch 2 and the plastidial phosphorylase 1 revealed protein-protein interactions with photosynthesis proteins in yeast two-hybrid screenings.

Muntaha S, Fettke J Plant Signal Behav. 2025; 20(1):2470775.

PMID: 40008471 PMC: 11866963. DOI: 10.1080/15592324.2025.2470775.

Development of Green Fluorescent Protein-Tagged Strains of via PEG-Mediated Genetic Transformation.

Ju F, Qi Z, Tan J, Dai T Microorganisms. 2025; 12(12.

PMID: 39770630 PMC: 11678020. DOI: 10.3390/microorganisms12122427.

Overexpression of NAC transcription factors from Eremopyrum triticeum promoted abiotic stress tolerance.

Zhong X, Peng J, Wang M, Yang X, Sun L Transgenic Res. 2024; 34(1):3.

PMID: 39738759 DOI: 10.1007/s11248-024-00428-3.

Phenotyping of a new yeast mapping population reveals differences in the activation of the TORC1 signalling pathway between wild and domesticated yeast strains.

Rocha G, Gomez M, Baeza C, Salinas F, Martinez C, Kessi-Perez E Biol Res. 2024; 57(1):82.

PMID: 39511644 PMC: 11545388. DOI: 10.1186/s40659-024-00563-5.

References

Ramon A, Fonzi W . Genetic transformation of Candida albicans. Methods Mol Biol. 2009; 499:169-74. DOI: 10.1007/978-1-60327-151-6_16. View

Tomlin G, Hamilton G, Gardner D, Walmsley R, Stateva L, Oliver S . Suppression of sorbitol dependence in a strain bearing a mutation in the SRB1/PSA1/VIG9 gene encoding GDP-mannose pyrophosphorylase by PDE2 overexpression suggests a role for the Ras/cAMP signal-transduction pathway in the control of yeast cell-wall.... Microbiology (Reading). 2000; 146 ( Pt 9):2133-2146. DOI: 10.1099/00221287-146-9-2133. View

Zheng H, Liu H, Chen S, Lu Z, Zhang Z, Pang D . Yeast transformation process studied by fluorescence labeling technique. Bioconjug Chem. 2005; 16(2):250-4. DOI: 10.1021/bc049833v. View

Engqvist-Goldstein A, Drubin D . Actin assembly and endocytosis: from yeast to mammals. Annu Rev Cell Dev Biol. 2003; 19:287-332. DOI: 10.1146/annurev.cellbio.19.111401.093127. View

Chen P, Liu H, Cui R, Zhang Z, Pang D, Xie Z . Visualized investigation of yeast transformation induced with Li+ and polyethylene glycol. Talanta. 2008; 77(1):262-8. DOI: 10.1016/j.talanta.2008.06.018. View

Tilburn J, Scazzocchio C, Taylor G, Lockington R, Davies R . Transformation by integration in Aspergillus nidulans. Gene. 1983; 26(2-3):205-21. DOI: 10.1016/0378-1119(83)90191-9. View

Becker D, Guarente L . High-efficiency transformation of yeast by electroporation. Methods Enzymol. 1991; 194:182-7. DOI: 10.1016/0076-6879(91)94015-5. View

Burgers P, Percival K . Transformation of yeast spheroplasts without cell fusion. Anal Biochem. 1987; 163(2):391-7. DOI: 10.1016/0003-2697(87)90240-5. View

Johnston S, Anziano P, Shark K, Sanford J, Butow R . Mitochondrial transformation in yeast by bombardment with microprojectiles. Science. 1988; 240(4858):1538-41. DOI: 10.1126/science.2836954. View

10.

Moreno S, Klar A, Nurse P . Molecular genetic analysis of fission yeast Schizosaccharomyces pombe. Methods Enzymol. 1991; 194:795-823. DOI: 10.1016/0076-6879(91)94059-l. View

11.

Cregg J, Barringer K, Hessler A, Madden K . Pichia pastoris as a host system for transformations. Mol Cell Biol. 1985; 5(12):3376-85. PMC: 369166. DOI: 10.1128/mcb.5.12.3376-3385.1985. View

12.

GURRIERI S, Wells K, Johnson I, Bustamante C . 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. 1997; 249(1):44-53. DOI: 10.1006/abio.1997.2102. View

13.

Klein R, Wolf E, Wu R, Sanford J . High-velocity microprojectiles for delivering nucleic acids into living cells. 1987. Biotechnology. 1992; 24:384-6. View

14.

Suga M, Hatakeyama T . A rapid and simple procedure for high-efficiency lithium acetate transformation of cryopreserved Schizosaccharomyces pombe cells. Yeast. 2005; 22(10):799-804. DOI: 10.1002/yea.1247. View

15.

Thompson J, Register E, Curotto J, Kurtz M, Kelly R . An improved protocol for the preparation of yeast cells for transformation by electroporation. Yeast. 1998; 14(6):565-71. DOI: 10.1002/(SICI)1097-0061(19980430)14:6<565::AID-YEA251>3.0.CO;2-B. View

16.

Okazaki K, Okazaki N, Kume K, Jinno S, Tanaka K, Okayama H . High-frequency transformation method and library transducing vectors for cloning mammalian cDNAs by trans-complementation of Schizosaccharomyces pombe. Nucleic Acids Res. 1990; 18(22):6485-9. PMC: 332599. DOI: 10.1093/nar/18.22.6485. View

17.

Costanzo M, Fox T . Transformation of yeast by agitation with glass beads. Genetics. 1988; 120(3):667-70. PMC: 1203545. DOI: 10.1093/genetics/120.3.667. View

18.

Boni L, Stewart T, Alderfer J, Hui S . Lipid-polyethylene glycol interactions: II. Formation of defects in bilayers. J Membr Biol. 1981; 62(1-2):71-7. DOI: 10.1007/BF01870201. View

19.

von Gersdorff K, Sanders N, Vandenbroucke R, De Smedt S, Wagner E, Ogris M . The internalization route resulting in successful gene expression depends on both cell line and polyethylenimine polyplex type. Mol Ther. 2006; 14(5):745-53. DOI: 10.1016/j.ymthe.2006.07.006. View

20.

Dawe A, Willins D, Morris N . Increased transformation efficiency of Aspergillus nidulans protoplasts in the presence of dithiothreitol. Anal Biochem. 2000; 283(1):111-2. DOI: 10.1006/abio.2000.4658. View