Electrotransformation of thermophilic bacterium Caldimonas manganoxidans
| Autor: | Takamitsu Arai, Kumar Sudesh, Toshiaki Kondo, Shimpei Aikawa, Akihiko Kosugi |
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| Rok vydání: | 2021 |
| Předmět: |
Microbiology (medical)
DNA Bacterial Chemistry Electroporation Thermophile Bacterial growth medicine.disease_cause Microbiology Comamonadaceae Transformation (genetics) chemistry.chemical_compound Plasmid Biochemistry medicine Transformation Bacterial Molecular Biology Escherichia coli DNA Transformation efficiency Plasmids |
| Zdroj: | Journal of microbiological methods. 192 |
| ISSN: | 1872-8359 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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