通过电穿孔法用广宿主范围质粒转化甲基营养型细菌——扭脱甲基杆菌。
Transformation of a methylotrophic bacterium, Methylobacterium extorquens, with a broad-host-range plasmid by electroporation.
作者信息
Ueda S, Matsumoto S, Shimizu S, Yamane T
机构信息
Department of Food Science and Technology, School of Agriculture, Nagoya University, Japan.
出版信息
Ann N Y Acad Sci. 1991 Dec 27;646:99-105. doi: 10.1111/j.1749-6632.1991.tb18568.x.
Electroporation was used to transform the methylotrophic bacterium Methylobacterium extorquens with broad-host-range plasmid pLA2917, which contains a gene specifying resistance to kanamycin. Plasmid DNA was introduced into M. extorquens in the presence of an electric pulse, and kanamycin-resistant transformants were obtained. These transformants harbored plasmid DNA that was identical to plasmid pLA2917. We examined several factors independently and found up to 8 x 10(3) transformants per microgram of DNA using 10 pulses with a duration of 300 microseconds at a field strength of 10 kV/cm.
采用电穿孔法，用含有卡那霉素抗性基因的广宿主质粒pLA2917转化甲基营养型细菌扭脱甲基杆菌。在电脉冲存在的情况下，将质粒DNA导入扭脱甲基杆菌，获得了卡那霉素抗性转化体。这些转化体携带的质粒DNA与质粒pLA2917相同。我们分别研究了几个因素，发现在场强为10 kV/cm、脉冲持续时间为300微秒、施加10个脉冲的条件下，每微克DNA可获得高达8×10³个转化体。
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