Sekizaki T, Tanoue T, Osaki M, Shimoji Y, Tsubaki S, Takai S
National Institute of Animal Health, Tsukuba, Ibaraki, Japan.
J Vet Med Sci. 1998 Feb;60(2):277-9. doi: 10.1292/jvms.60.277.
The condition of an electroporation method was re-evaluated for the introduction of foreign plasmid DNA into Rhodococcus equi. The method is based on an electroporation of the bacteria made competent by culturing in a broth containing glycine and by heat shock at 50 degrees C. Transformation of R. equi could be achieved with a chloramphenicol-resistant shuttle vector originating from Rhodococcus fascians at an efficiency of about 10(4) transformants/microgram DNA. The bacteria were also shown to become competent when they were incubated with a chemical transformation buffer prior to washing with an electroporation buffer.
重新评估了电穿孔法将外源质粒DNA导入马红球菌的条件。该方法基于在含有甘氨酸的肉汤中培养并经50℃热休克使细菌变得有感受态后进行电穿孔。用源自筋膜红球菌的氯霉素抗性穿梭载体可实现马红球菌的转化，转化效率约为10⁴个转化子/微克DNA。还表明，在用电穿孔缓冲液洗涤之前，将细菌与化学转化缓冲液一起孵育时，它们会变得有感受态。
