Abstract
Electroporation is a method for the introduction of molecules (usually nucleic acids) into a cell, consisting of submitting the cells to high-voltage and short electric pulses in the presence of the exogenous DNA/molecule. It is a versatile method, adaptable to different types of cells, from bacteria to cultured cells to higher eukaryotes, and thus has applications in many diverse fields, such as environmental biology, biotechnology, genetic engineering, and medicine. Electroporation has some advantages over other genetic transformation strategies, including the simplicity of the method, a wide range of adjustable parameters (possibility of optimization), high reproducibility and avoidance of the use of chemicals toxic to cells. Here we describe an optimized electroporation procedure for the industrially important fungus Acremonium chrysogenum, using germinated conidia and fragmented young mycelium. In both cases, the transformation efficiency was higher compared to the conventional polyethylene glycol (PEG)-mediated transformation of protoplasts.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Cohen SN, Chang AC, Hsu L (1972) Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A 69:2110–2114
Ruiz-Diez B (2002) Strategies for the transformation of filamentous fungi. J Appl Microbiol 92:189–195
Chakraborty BN (2014) Electroporation mediated DNA transformation of filamentous fungi. In: van den Berg MA, Maruthachalam K (eds) Genetic transformation systems in fungi, vol 1. Springer International Publishing, Cham, pp 67–79
Hashimoto H, Morikawa H, Yamada Y, Kimura A (1985) A novel method for transformation of intact yeast cells by electroinjection of plasmid DNA. Appl Microbiol Biotechnol 21:336–339
Delorme E (1989) Transformation of Saccharomyces cerevisiae by electroporation. Appl Environ Microbiol 55:2242–2246
Richey MG, Marek ET, Schardl CL, Smith DA (1989) Transformation of filamentous fungi with plasmid DNA by electroporation. Phytopathology 79:844–847
Tsai HF, Siegel MR, Schardl CL (1992) Transformation of Acremonium coenophialum, a protective fungal symbiont of the grass Festuca arundinacea. Curr Genet 22:399–406
Chakraborty BN, Patterson NA, Kapoor M (1991) An electroporation-based system for high-efficiency transformation of germinated conidia of filamentous fungi. Can J Microbiol 37:858–863
Sánchez O, Aguirre J (1996) Efficient transformation of Aspergillus nidulans by electroporation of germinated conidia. Fungal Genet Newslett 43:48–51
Robinson M, Sharon A (1999) Transformation of the bioherbicide Colletotrichum gloeosporioides f. sp. aeschynomene by electroporation of germinated conidia. Curr Genet 36:98–104
Xu S, Zhou Z, Du G, Zhou J, Chen J (2014) Efficient transformation of Rhizopus delemar by electroporation of germinated spores. J Microbiol Methods 103:58–63
Kuo CY, Huang CT (2008) A reliable transformation method and heterologous expression of beta-glucuronidase in Lentinula edodes. J Microbiol Methods 72:111–115
Pigac J, Schrempf H (1995) A simple and rapid method of transformation of Streptomyces rimosus R6 and other Streptomycetes by electroporation. Appl Environ Microbiol 61:352–356
Jiang Q, Ying SH, Feng MG (2007) Enhanced frequency of Beauveria bassiana blastospore transformation by restriction enzyme mediated integration and electroporation. J Microbiol Methods 69:512–517
Miklenić M, Štafa A, Bajić A, Žunar B, Lisnić B, Svetec IK (2013) Genetic transformation of the yeast Dekkera/Brettanomyces bruxellensis with non-homologous DNA. J Microbiol Biotechnol 23:674–680
Díaz A, Villanueva P, Oliva V, Gil-Duran C, Fierro F, Chávez R, Vaca I (2019) Genetic transformation of the filamentous fungus Pseudogymnoascus verrucosus of Antarctic origin. Front Microbiol 10:2675
Kawai S, Hashimoto W, Murata K (2010) Transformation of Saccharomyces cerevisiae and other fungi: methods and possible underlying mechanism. Bioeng Bugs 1:395–403
Cruz-Ramón J, Fernández FJ, Mejía A, Fierro F (2018) Electroporation of germinated conidia and young mycelium as an efficient transformation system for Acremonium chrysogenum. Folia Microbiol 64:33–39
Nash CH, Pieper RL (1974) Physiology of spore germination in Cephalosporium acremonium. Mycopathol Mycol Appl 54:369–375
Chakraborty BN, Kapoor M (1990) Transformation of filamentous fungi by electroporation. Nucleic Acids Res 18:6737
Skatrud PL, Queener SW, Carr LG, Fisher DL (1987) Efficient integrative transformation of Cephalosporium acremonium. Curr Genet 12:337–348
Acknowledgments
This work was funded by the CONACyT (México) through the Research Project CB-2008-01 105527. Jessica Cruz-Ramón received a Scholarship Grant from the CONACyT (No. 203440).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Cruz-Ramón, J., Fernández, F.J., Fierro, F. (2021). High-Efficiency Electroporation for Genetic Improvement of Fungal Strains. In: Barreiro, C., Barredo, JL. (eds) Antimicrobial Therapies. Methods in Molecular Biology, vol 2296. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1358-0_10
Download citation
DOI: https://doi.org/10.1007/978-1-0716-1358-0_10
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1357-3
Online ISBN: 978-1-0716-1358-0
eBook Packages: Springer Protocols