Stable transformants of Chromobacterium violaceum were obtained by high-voltage electroporation with a 7-kilobase binary plasmid. The technique was reliable, reproducible, and simple, with efficiencies of 105 transformants/μg of plasmid DNA. The electrical conditions that resulted in the highest efficiencies were short pulse length (4.4–4.5 ms) and high voltage (12.5 kV/cm). The numbers of transformants were almost the same during the growth exponential phase (variation at optical density) and resulted in the highest efficiencies at DNA concentration of 250 pg/ml. Saturation appeared to begin at 4 μg/ml of DNA. This method of C. violaceum transformation should enhance the genetic and biotechnological research by providing a valuable, widely used procedure of introducing DNA into this bacterium.
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We thank S.C. da Silva and A. Schrank for critically reading this manuscript. This work was supported by grants from Brazilian National Research Council-CNPq/MCT-Proc 680220/00-5 and Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS). L.B, R.C., and S.W. are recipients of scholarships from CNPq (Functional Genome) and F.H.S. from FAPERGS.
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Broetto, L., Cecagno, R., Sant'Anna, F.H. et al. Stable transformation of Chromobacterium violaceum with a broad-host-range plasmid. Appl Microbiol Biotechnol 71, 450–454 (2006). https://doi.org/10.1007/s00253-005-0140-5
- Transformation Efficiency
- Pulse Length
- High Transformation Efficiency
- Chromobacterium Violaceum