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Agrobacterium tumefaciens-mediated transformation of antifungal lipopeptide producing fungus Coleophoma empetri F-11899

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Abstract

The filamentous fungus Coleophoma empetri F-11899 produces an echinocandin-like compound FR901379, the original source for micafungin which is prescribed to treat deep-seated mycoses. Despite its industrial importance, no genetic information on C. empetri F-11899 is currently available. To characterize FR901379 biosynthetic genes by insertional mutagenesis and to improve the compound production genetically, Agrobacterium tumefaciens-mediated transformation (ATMT) was attempted to make genetic manipulation possible in this strain. The optimum conditions for ATMT of C. empetri were determined for the cell density of bacteria, time period of co-cultivation and types of filters in co-cultivation. Using the established ATMT method, the hygromycin B resistant gene was successfully transferred into the genome of C. empetri F-11899 and stably maintained even after a serial passage. Some of these results will be applicable for ATMT of various filamentous fungi.

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Acknowledgments

We are grateful to Dr. Kang (Pennsylvania State University, PA, USA), for providing the pBHt2 plasmid. This research was supported by a Grant-in-Aid for Young Scientists (B), (21780079) from The Ministry of Education, Culture, Sports, Science and Technology in Japan (MEXT).

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Authors and Affiliations

  1. Department of Biotechnology, Faculty of Engineering, Toyama Prefectural University, 5180 Kurokawa, Imizu, Toyama, 939-0398, Japan

    Masato Yamada, Kazunobu Yawata, Yohsuke Orino, Yasuhiro Isogai & Seiji Hashimoto

  2. Fermentation and Biotechnology Laboratories, Astellas Pharm Inc., 156 Nakagawara, Kiyosu, Aichi, 452-0915, Japan

    Satoshi Ueda

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

    Goro Taguchi & Makoto Shimosaka

  4. Biotechnology Research Center, Toyama Prefectural University, 5180 Kurokawa, Imizu, Toyama, 939-0398, Japan

    Seiji Hashimoto

Authors
  1. Masato Yamada
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  2. Kazunobu Yawata
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  3. Yohsuke Orino
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  4. Satoshi Ueda
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  5. Yasuhiro Isogai
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  6. Goro Taguchi
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  7. Makoto Shimosaka
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  8. Seiji Hashimoto
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Corresponding authors

Correspondence to Masato Yamada or Seiji Hashimoto.

Additional information

Communicated by J. Heitman.

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Yamada, M., Yawata, K., Orino, Y. et al. Agrobacterium tumefaciens-mediated transformation of antifungal lipopeptide producing fungus Coleophoma empetri F-11899. Curr Genet 55, 623–630 (2009). https://doi.org/10.1007/s00294-009-0275-5

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  • DOI: https://doi.org/10.1007/s00294-009-0275-5

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