地衣芽孢杆菌感受态细胞的诱导形成及其高效电转化方法

Formation of Competent Bacillus licheniformis Cell and High Efficiency Electrotransformation for Vectors

通过体外处理诱导地衣芽孢杆菌产生感受态性能 ,同时调整参数 ,建立了感受态细胞对质粒 pAPR的高效电转化方法 .当质粒DNA为 1.5 μg/mL、转化时电压为175 0V时 ,可得到 2 6 1个转化子 ,经鉴定均为阳性克隆子 .此为以芽孢杆菌为宿主进行高效电转化及为建立适合工业应用的分泌型表达载体提供参考 .

Bacillus licheniformis was induced to form competent cells in vitro, and by changing parameters, a method of high efficiency electrotransformation for plasmid pAPR was established. When DNA concentration was 1.5 μg/mL and the voltage was 1 750 V, 261 transformants were obtained on screen plates. By the identification, all of transformants were positive clones. With ordinary electrotransformation method, 20 transformants were obtained on screen plates. The experiments provided a model for high efficiency electrotransformation with Bacillus strains as host cells, and constructed a foundation for looking for adaptable secretory expression vectors in industry.