Stable transformation of Chromobacterium violaceum with a broad-host-range plasmid |
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Authors: | L Broetto R Cecagno F H Sant'Anna S Weber I S Schrank |
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Institution: | 1. Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Av. Bento Gon?alves, 9500-Prédio 43421, C.P. 15005-CEP: 91501-970, Porto Alegre, RS, Brazil 2. Depto. de Biologia Molecular e Biotecnologia, UFRGS, Porto Alegre, Brazil
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Abstract: | 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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