多杀性巴氏杆菌C48-16电转化条件的研究

Study on Optional Conditions of Electroporation in Pasteurella Multocida C48-16

将C48-16感受态细胞和带有标记基因的质粒DNA混和后进行电转化,通过改变脉冲电压、脉冲次数和质粒DNA浓度等电转化的重要参数,探讨了巴氏杆菌(Pasteurella multocida)C48-16的电转化条件。结果显示:最佳脉冲电压为2.4～2.5kV,最佳脉冲次数为1～2。质粒DNA浓度为58～1.16×102μg/mL转化子最多。所有转化子经标记基因PCR鉴定为阳性克隆子。此方法与常规化学转化法相比操作简单、快速且转化成功率更高。

To approach the conditions of electroporation of Pasteurella multocida C48-16,the mixture of C48-16 competent cells and plasmid DNA with label gene were electrotransfected,by changing some important effecting factors,such as voltage,pulse plength,concentration of plasmid and so on.The results showed that many transformants were obtained in the best screen plate when the voltage was set from 2.4 to 2.5 kV,pulses was 1 to 2.And the optimum concentration of plasmid DNA was 58～1.16×102 μg/mL. By the identification,all transformants were positive clones.Compared with ordinary chemical transfecting method,electroporation is more easy and rapid in manipulation,furthermore has higher transfecting efficiency.