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Generation of readily transformable Bacillus licheniformis mutants

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Abstract

A set of mutants was generated by targeted deletion of the hsdR loci of two type I restriction modification systems (RMS) identified in Bacillus licheniformis DSM13. Single as well as double knock-outs resulted in strains being readily transformable with plasmids isolated from Bacilli. Introduction of shuttle plasmids isolated from Escherichia coli was routinely possible when the double mutant B. licheniformis MW3 (ΔhsdR1, ΔhsdR2) was used in transformation experiments. Growth and secretion of extracellular enzymes were not affected in any of the mutants. Thus, along with an optimized transformation protocol, this study makes available an urgently needed transformation system for this industrially exploited species.

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Acknowledgement

We gratefully acknowledge encouraging comments from a number of colleagues, which already used successfully strain MW3 in their transformation experiments. The work was supported by the Federal Ministry of Education and Research (BMBF, Bonn-Bad Godesberg, Germany) grant no. 0313645.

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Authors and Affiliations

  1. Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität Münster, Corrensstrasse 3, 48149, Münster, Germany

    Bianca Waschkau, Jens Waldeck & Friedhelm Meinhardt

  2. Henkel KgaA, VTB-Enzymtechnologie, Henkelstrasse 67, 40191, Düsseldorf, Germany

    Susanne Wieland & Renèe Eichstädt

Authors
  1. Bianca Waschkau
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  2. Jens Waldeck
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  3. Susanne Wieland
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  4. Renèe Eichstädt
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  5. Friedhelm Meinhardt
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Corresponding author

Correspondence to Friedhelm Meinhardt.

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This work is dedicated to Prof. Dr. Hans Jürgen Rehm on the occasion of his 80th birthday.

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Waschkau, B., Waldeck, J., Wieland, S. et al. Generation of readily transformable Bacillus licheniformis mutants. Appl Microbiol Biotechnol 78, 181–188 (2008). https://doi.org/10.1007/s00253-007-1278-0

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  • DOI: https://doi.org/10.1007/s00253-007-1278-0

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