Affiliations 

  • 1 Japan International Research Center for Agricultural Sciences, 1-1 Ohwashi Tsukuba, Ibaraki 305-8686, Japan
  • 2 School of Biological Sciences, Universiti Sains Malaysia, 11800 Penang, Malaysia
  • 3 Japan International Research Center for Agricultural Sciences, 1-1 Ohwashi Tsukuba, Ibaraki 305-8686, Japan. Electronic address: akosugi@affrc.go.jp
J Microbiol Methods, 2022 01;192:106375.
PMID: 34793853 DOI: 10.1016/j.mimet.2021.106375

Abstract

Caldimonas manganoxidans is a Gram-negative, thermophilic, bioplastic-producing bacterium that is a promising strain to overcome the drawbacks of existing bioplastic manufacturing methods. However, genetic manipulation of this species has not previously been studied. Here, we developed an optimized electrotransformation protocol for C. manganoxidans by screening conditions, including the bacterial growth phase, electroporation buffer, pulse strength, and recovery time. The optimized transformation protocol obtained (3.1 ± 0.78) × 108 colony-forming units/μg DNA of plasmid pBBR1MCS-2. High transformation efficiency was observed when using plasmid DNA isolated from C. manganoxidans. The DNA methylases of Escherichia coli did not affect the transformation efficiency of C. manganoxidans. The electrotransformation technique proposed here will be beneficial for the genetic manipulation of thermophilic Caldimonas species.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.