Mahony D E, Mader J A, Dubel J R
Department of Microbiology, Faculty of Medicine, Dalhousie University, Halifax, Nova Scotia, Canada.
Appl Environ Microbiol. 1988 Jan;54(1):264-7. doi: 10.1128/aem.54.1.264-267.1988.
L-form (L-phase) cultures of Clostridium perfringens were tested for their transformability with plasmid DNA. Three L-form strains were transformable, but one, strain L-13, was superior to the others. This strain was easily and reproducibly transformed with previously described shuttle vectors which were derived from either C. perfringens or Escherichia coli. Strain L-13 was transformable by a variety of methods, and a new micromethod worked well under both aerobic and anaerobic conditions. The maximal number of transformants was attained after strain L-13 was exposed for 4 h to the transforming DNA and polyethylene glycol. Viable counts determined in tubes of semisolid brain heart infusion medium containing 10% sucrose, with or without 2 micrograms of tetracycline per ml, showed a transformation rate of 3.9 X 10(-5) (transformants per viable cells).
对产气荚膜梭菌的L型(L相)培养物进行了质粒DNA转化能力测试。三株L型菌株具有可转化性,但其中一株L-13菌株比其他菌株更具优势。该菌株很容易且可重复地被先前描述的穿梭载体转化,这些穿梭载体来源于产气荚膜梭菌或大肠杆菌。L-13菌株可通过多种方法进行转化,一种新的微量方法在需氧和厌氧条件下均效果良好。L-13菌株在暴露于转化DNA和聚乙二醇4小时后获得了最大数量的转化子。在含有10%蔗糖、每毫升含或不含2微克四环素的半固体脑心浸液培养基管中测定的活菌数显示,转化率为3.9×10^(-5)(每活细胞的转化子数)。