Bacterial transformation is a crucial step in the genetic manipulation of a bacterium. However, Gram-positive bacteria are difficult to transform and consequently many different methodologies have been developed. Here, we examined the transformation efficiencies of an electroporation protocol by varying three main factors: the composition of the electroporation buffer, the strength of the electric pulse, and the composition of the recovery media. Overall, transformation efficiency was enhanced when we prepared the electrocompetent cells at room temperature instead of an ice-cold temperature. The protocol detailed in this work was demonstrated to be applicable to another B. cereus strain and two other Bacillus species, and has the potential to be applied to other undomesticated Gram-positive and/or rhizospheric bacterial strains that are difficult to transform using current methodologies.

中文翻译：

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使用室温电感受态细胞制备方案转化根际芽孢杆菌B25菌株。

细菌转化是细菌基因操作中的关键步骤。然而，革兰氏阳性细菌难以转化，因此已经开发出许多不同的方法。在这里，我们通过改变三个主要因素来检查电穿孔方案的转化效率：电穿孔缓冲液的组成，电脉冲的强度和回收介质的组成。总的来说，当我们在室温而不是冰冷的温度下制备电感受态电池时，转化效率得到了提高。这项工作中详述的方案已证明适用于另一种蜡状芽孢杆菌菌株和另外两种芽孢杆菌属物种，