Morikawa M, Daido H, Pongpobpibool S, Imanaka T
Department of Biotechnology, Faculty of Engineering, Osaka University, Japan.
Appl Microbiol Biotechnol. 1994 Nov;42(2-3):300-3. doi: 10.1007/BF00902732.
Arthrobacter sp. strain MIS38 was transformed with a shuttle vector containing the kanamycin resistant gene kan (derived from Tn5) by an electroporation method. This shuttle vector is from Brevibacterium lactofermentum and Escherichia coli, pULRS8. The following optimal condition of electroporation was determined. A square wave pulse of 1 kV/cm electric field strength for 0.5 ms duration yielded 3 x 10(5) transformants/micrograms plasmid DNA. The number of transformants increased with the amount of DNA over the range 0.01-5 micrograms. This host-vector system was then used successfully to clone and express a lipase gene from Arthrobacter sp. strain MIS38 into both Arthrobacter sp. MIS38 and E. coli JM109.
通过电穿孔法，用含有卡那霉素抗性基因kan（源自Tn5）的穿梭载体pULRS8（来自乳酸发酵短杆菌和大肠杆菌）转化节杆菌属菌株MIS38。确定了以下电穿孔的最佳条件。1 kV/cm电场强度、持续时间为0.5 ms的方波脉冲可产生3×10⁵个转化子/微克质粒DNA。在0.01 - 5微克的范围内，转化子数量随DNA量的增加而增加。然后成功地利用这个宿主 - 载体系统将节杆菌属菌株MIS38的脂肪酶基因克隆并表达至节杆菌属MIS38和大肠杆菌JM109中。
