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Methods for the genetic manipulation of Nonomuraea sp. ATCC 39727

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Journal of Industrial Microbiology & Biotechnology

Abstract

Nonomuraea sp. ATCC 39727 belongs to the Streptosporangiaceae family of filamentous actinomycetes. This microorganism produces the teicoplanin-like glycopeptide A40926, which is the starting material for the synthesis of the second-generation glycopeptide dalbavancin. Notwithstanding the strain’s pharmaceutical relevance, the lack or poor efficiency of genetic tools to manipulate Nonomuraea sp. ATCC 39727 has hampered strain and product improvement. Here we report the development of gene transfer systems based on protoplast transformation and intergeneric conjugation from Escherichia coli. Efficiency of transformation and conjugation, followed by site specific or homologous recombination with the Nonomuraea sp. genome, were determined using the integrative plasmid pSET152 (5.7 kb), and the Supercos1 derivative cosmid A40ΔY (30 kb). To our knowledge, this is the first report of the transformation of protoplasts of Nonomuraea sp. ATCC 39727, even though the improved procedure for intergeneric conjugation makes it the method of choice for introducing large segments of DNA into Nonomuraea sp. ATCC 39727.

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Acknowledgments

This work was supported by FAR 2007–2008–2009 to F.M. and by MIUR fellowship to G.L.M. We thank also the support from Consorzio Interuniversitario per le Biotecnologie (CIB) and Progetto Cariplo: Promuovere Capitale Umano d’Eccellenza, for the grants to G.L.M. and E.B. L.F. and M.B. were funded by grants from the Biotechnology and Biological Sciences Research Council, U.K.

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Authors and Affiliations

  1. Dipartimento di Biotecnologie e Scienze Molecolari, Università degli Studi dell’Insubria, via J. H. Dunant 3, Varese, 21100, Italy

    Giorgia Letizia Marcone, Elisa Binda & Flavia Marinelli

  2. Centro Interuniversitario di Ricerca in Biotecnologie Proteiche “The Protein Factory”, Politecnico di Milano and Università degli Studi dell’Insubria, Varese, Italy

    Giorgia Letizia Marcone, Elisa Binda & Flavia Marinelli

  3. John Innes Centre, Norwich Research Park, Colney Lane, Norwich, NR4 7UH, UK

    Lucy Foulston & Mervyn Bibb

  4. Actygea S.r.l., via R. Lepetit 34, 21040, Gerenzano (VA), Italy

    Fabrizio Beltrametti

Authors
  1. Giorgia Letizia Marcone
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  2. Lucy Foulston
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  3. Elisa Binda
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  4. Flavia Marinelli
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  5. Mervyn Bibb
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  6. Fabrizio Beltrametti
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Corresponding author

Correspondence to Giorgia Letizia Marcone.

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Marcone, G.L., Foulston, L., Binda, E. et al. Methods for the genetic manipulation of Nonomuraea sp. ATCC 39727. J Ind Microbiol Biotechnol 37, 1097–1103 (2010). https://doi.org/10.1007/s10295-010-0807-5

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  • DOI: https://doi.org/10.1007/s10295-010-0807-5

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