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Applied Microbiology and Biotechnology

, Volume 101, Issue 2, pp 735–747 | Cite as

Magnesium aminoclay-based transformation of Paenibacillus riograndensis and Paenibacillus polymyxa and development of tools for gene expression

  • Luciana Fernandes Brito
  • Marta Irla
  • Tatjana Walter
  • Volker F. Wendisch
Methods and protocols

Abstract

Members of the genus Paenibacillus are widespread facultative anaerobic, endospore-forming bacteria. Some species such as Paenibacillus riograndensis or Paenibacillus polymyxa fix nitrogen and may play an important role in agriculture to reduce mineral nitrogen fertilization in particular for non-legume plants. The genetic manipulation of Paenibacillus is an imperative for the functional characterization, e.g., of its plant growth-promoting activities and metabolism. This study showed that P. riograndensis and P. polymyxa can be readily transformed using physical permeation by magnesium aminoclays. By means of the fluorescent reporter genes gfpUV, mcherry, and crimson, a two-plasmid system consisting of a theta-replicating plasmid and a rolling circle-replicating plasmid was shown to operate in both species. Xylose-inducible and mannitol-inducible fluorescent reporter gene expression was demonstrated in the compatible two-plasmid system by fluorescence-activated cell scanning. As a metabolic engineering application, the biotin requiring P. riograndensis was converted to a biotin-prototrophic strain based on mannitol-inducible expression of the biotin biosynthesis operon bioWAFDBI from Bacillus subtilis.

Keywords

Paenibacillus Theta-replicating plasmid Magnesium aminoclays Transformation Biotin prototrophy 

Notes

Acknowledgements

L.F.B. acknowledges financial support from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Science without Borders program), Brazil. M.I. acknowledges support from the CLIB Graduate Cluster Industrial Biotechnology at Bielefeld University, Germany which is financed by a grant from the Federal Ministry of Innovation, Science and Research (MIWF) of the federal state North Rhine-Westphalia, Germany.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

The research performed did not involve human participants and/or animals.

Supplementary material

253_2016_7999_MOESM1_ESM.pdf (303 kb)
ESM 1 (PDF 303 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Luciana Fernandes Brito
    • 1
  • Marta Irla
    • 1
  • Tatjana Walter
    • 1
  • Volker F. Wendisch
    • 1
  1. 1.Genetics of Prokaryotes, Faculty of Biology and Center for Biotechnology (CeBiTec)Bielefeld UniversityBielefeldGermany

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