Development of an efficient electroporation method for rhizobacterial Bacillus mycoides strains

Yanglei Yi; Oscar P Kuipers

doi:10.1016/j.mimet.2016.12.022

Development of an efficient electroporation method for rhizobacterial Bacillus mycoides strains

J Microbiol Methods. 2017 Feb:133:82-86. doi: 10.1016/j.mimet.2016.12.022. Epub 2016 Dec 30.

Authors

Yanglei Yi¹, Oscar P Kuipers²

Affiliations

¹ Molecular Genetics, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, The Netherlands.
² Molecular Genetics, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, The Netherlands. Electronic address: o.p.kuipers@rug.nl.

PMID: 28042055
DOI: 10.1016/j.mimet.2016.12.022

Abstract

In order to develop a method for electroporation of environmental Bacillus mycoides strains, we optimized several conditions that affect the electroporation efficiency of this bacterium. By combining the optimized conditions, the electroporation efficiency of strain EC18 was improved to (1.3±0.6)×10⁵cfu/μg DNA, which is about 10³-fold increase in comparison with a previously reported value. The method was further validated on various B. mycoides strains, yielding reasonable transformation efficiencies. Furthermore, we confirmed that restriction/modification is the main barrier for electroporation of this bacterium. To the best of our knowledge, this is the first systematic investigation of various parameters of electroporation of B. mycoides. The electroporation method reported will allow for efficient genetic manipulation of this bacterium.

Keywords: Bacillus mycoides; Electroporation; Optimization; Transformation.

MeSH terms

Bacillus / genetics*
Bacillus / growth & development
Culture Media / chemistry
DNA, Bacterial / genetics
Electroporation / methods*
Genetic Techniques
Plasmids / genetics

Substances

Culture Media
DNA, Bacterial