Broetto L, Cecagno R, Sant'anna F H, Weber S, Schrank I S
Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves, 9500-Prédio 43421, C.P. 15005-CEP: 91501-970, Porto Alegre, RS, Brazil.
Appl Microbiol Biotechnol. 2006 Jul;71(4):450-4. doi: 10.1007/s00253-005-0140-5. Epub 2005 Sep 20.
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 10(5) transformants/microg 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 microg/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.
通过用一个7千碱基的二元质粒进行高压电穿孔获得了紫色色杆菌的稳定转化体。该技术可靠、可重复且简单，质粒DNA的转化效率为10(5)个转化体/微克。导致最高效率的电参数是短脉冲长度（4.4 - 4.5毫秒）和高电压（12.5千伏/厘米）。在生长指数期（光密度变化）转化体数量几乎相同，在DNA浓度为250皮克/毫升时效率最高。DNA浓度为4微克/毫升时似乎开始出现饱和。这种紫色色杆菌转化方法通过提供一种有价值且广泛应用的将DNA导入该细菌的程序，应能加强遗传学和生物技术研究。
Zotero 插件