Bonnassie S, Burini J F, Oreglia J, Trautwetter A, Patte J C, Sicard A M
Centre de Recherche de Biochimie et de Génétique Cellulaires du CNRS, Toulouse, France.
J Gen Microbiol. 1990 Oct;136(10):2107-12. doi: 10.1099/00221287-136-10-2107.
The Escherichia coli-Brevibacterium lactofermentum shuttle vector pBLA was introduced into intact cells of B. lactofermentum by electrotransformation. Several parameters of this procedure such as voltage and cell concentration were analysed. Optimal conditions gave an efficiency of 10(6) transformants per microgram of DNA. Two recalcitrant strains could be electrotransformed when an ampicillin pretreatment step was used. Electrotransformation experiments using DNAase or different structural forms of plasmid DNA showed that the electrotransformation process is quite different from natural transformation involving competence development. Restriction-modification-proficient B. lactofermentum could be efficiently electrotransformed with pBLA DNA isolated from E. coli. This restriction-modification system therefore seems to be overcome by electrotransformation. Thus electrotransformation may efficiently replace the protoplast bacterial transformation method.
通过电转化将大肠杆菌-乳酸发酵短杆菌穿梭载体pBLA导入乳酸发酵短杆菌的完整细胞中。分析了该过程的几个参数，如电压和细胞浓度。最佳条件下每微克DNA的转化效率为10(6)个转化体。当使用氨苄青霉素预处理步骤时，两种难转化的菌株可以被电转化。使用DNA酶或不同结构形式的质粒DNA进行的电转化实验表明，电转化过程与涉及感受态发育的自然转化有很大不同。从大肠杆菌中分离得到的pBLA DNA可以有效地电转化具有限制-修饰功能的乳酸发酵短杆菌。因此，这种限制-修饰系统似乎可以通过电转化被克服。因此，电转化可能有效地替代原生质体细菌转化方法。
