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pFungiway: a series of plasmid vectors used for gene manipulation in fungi

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Abstract

A series of plasmid vectors, named pFungiway, was constructed for the purpose of simple and versatile gene manipulation in a wide range of fungi. The backbone of these vectors consists of the binary plasmid pCAMBIA2200, which can be introduced into fungal cells by Agrobacterium tumefaciens-mediated transformation. Objective genes can be easily inserted into the vector by the use of Gateway technology, without relying on any restriction cleavage sites. The pFungiway vectors are composed of two types: expression vectors, in which the expression of the objective genes is driven by a constitutive promoter; and repression vectors, in which the expression of the endogenous target gene is repressed by RNA interference. Either of two genes conferring resistance to hygromycin B or G418 can be used as a selective marker. The availability of pFungiway vectors was confirmed by the use of fluorescent reporter genes in the basidiomycete Flammulina velutipes and in the plant pathogenic fungus Fusarium oxysporum.

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Acknowledgments

This work was supported by a Grant-in-Aid for Scientific Research (C) (19580086) from the Japan Society for the Promotion of Science (JSPS). We are indebted to the Division of Gene Research, Research Center for Human and Environmental Sciences, Shinshu University, for providing facilities.

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Authors and Affiliations

  1. Division of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, 3-15-1 Tokida, Ueda, Nagano, 386-8567, Japan

    Ryohei Nishikawa, Masumi Yoshida, Tomotsugu Noda, Toru Okuhara, Goro Taguchi & Makoto Shimosaka

  2. Mushroom Research Laboratory, Hokuto Corporation, 800-8 Shimokomazawa, Nagano, 381-0008, Japan

    Satoshi Inatomi

Authors
  1. Ryohei Nishikawa
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  2. Masumi Yoshida
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  3. Tomotsugu Noda
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  4. Toru Okuhara
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  5. Goro Taguchi
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  6. Satoshi Inatomi
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  7. Makoto Shimosaka
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Corresponding author

Correspondence to Makoto Shimosaka.

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Nishikawa, R., Yoshida, M., Noda, T. et al. pFungiway: a series of plasmid vectors used for gene manipulation in fungi. Ann Microbiol 66, 825–832 (2016). https://doi.org/10.1007/s13213-015-1166-2

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  • DOI: https://doi.org/10.1007/s13213-015-1166-2

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