地衣芽孢杆菌感受态细胞的形成及高效电转化

Formation of competent Bacillus licheniformis cell and high efficiency electrotrans formation for vectors

芽孢杆菌在营养缺乏的饥饿状态下 ,细胞易产生感受态因子 ,处于生长芽孢时期的芽孢杆菌更容易产生感受态。基于此原则利用芽孢杆菌极限营养培养基通过体外处理诱导使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浓度为 1 .5μg/ml、转化时电压为 1 750V的时候 ,可以得到 2 6 1个转化子 ,经鉴定均为阳性克隆子。而常规电转化的最高仅为 2 0个转化子。为以芽孢杆菌为宿主进行高效电转化提供了模型 。

Bacillus licheniformis was induced to form competent cell in vitro , and by changing parameters, the method of high efficiency electrotransformation for plasmid pAPR was constructed. When DNA concentration was 1.5 μg/ml and the voltage was 1750 V, 261 transformats were obtained in screen plate.By the identification, all of transformants were positive clones.With ordinary electrotransformation method, 20 transformants were obtaind in screen plate. The experiment 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.