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Comparison between polyethylene glycol- and polyethylenimine-mediated transformation of Aspergillus nidulans

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Abstract

Genetic transformation of many filamentous fungi is carried out by a protocol that utilizes polyethylene glycol (PEG) and calcium ion (Ca2+). This method has remained practically unchanged for more than 20 years, but the roles these molecules play are not definitively understood. To gain a better understanding, we have compared PEG transformation to a protocol using polyethylenimine (PEI) that is the basis for non-viral transfection in mammals and which has a well established molecular model for assisting DNA uptake. Protoplasts of Aspergillus nidulans could be transformed in the presence of Ca2+ with a relatively high ratio of PEI to DNA molecules. By comparing PEI and PEG in terms of interaction with DNA, fungal protoplasts, and response to different transformation conditions, we propose that the role of PEG is most likely to function after transforming DNA is incorporated into protoplasts, rather than the accepted view that it functions outside of the cell. Confirmation that protoplast fusion was not involved in DNA uptake is consistent with this hypothesis.

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Acknowledgments

We are grateful to Dr. Austen Ganley at National Institute of Genetics, Japan, and Dr. Richard Johnson at AgResearch Grasslands, New Zealand for critical readings of the manuscript. Our appreciation is also forwarded to Dr. Mikihiro Yamamoto in Okayama University and Dr. Motoichiro Kodama in Tottori University, Japan, for their critical help in the course of this study.

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Author notes

  1. Tetsuya Kuwano

    Present address: Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Minato-ku, Tokyo, 108-8639, Japan

Authors and Affiliations

  1. Department of Biological Sciences, Faculty of Science, Shinshu University, Matsumoto, Nagano, 390-8621, Japan

    Tetsuya Kuwano & Chikako Shirataki

  2. Division of Science and Technology, Graduate School of Shinshu University, Matsumoto, Nagano, 390-8621, Japan

    Yasuo Itoh

Authors
  1. Tetsuya Kuwano
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  2. Chikako Shirataki
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  3. Yasuo Itoh
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Correspondence to Yasuo Itoh.

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Communicated by Gerhard Braus.

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Kuwano, T., Shirataki, C. & Itoh, Y. Comparison between polyethylene glycol- and polyethylenimine-mediated transformation of Aspergillus nidulans . Curr Genet 54, 95–103 (2008). https://doi.org/10.1007/s00294-008-0204-z

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