Luchansky J B, Muriana P M, Klaenhammer T R
Department of Food Science, North Carolina State University, Raleigh 27695-7624.
Mol Microbiol. 1988 Sep;2(5):637-46. doi: 10.1111/j.1365-2958.1988.tb00072.x.
Plasmid DNA was introduced by electroporation into Bacillus, Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Listeria, Pediococcus, Propionibacterium and Staphylococcus as an alternative to competent-cell or protoplast transformation. Plasmid-containing transformants were recovered in these recipients at frequencies ranging from 10(1) to 10(5) transformants micrograms-1 of pGK12. Several parameters of the protocol, including DNA concentration, voltage, plating regimen and electroporation buffers were evaluated to determine conditions that improved transformation frequencies for Lactobacillus acidophilus. Using optimized conditions, the following plasmids were introduced into L. acidophilus: pAMB1, pC194, pGB354, pGKV1, pSA3, pTRK13, pTV1 and pVA797. The ability to transfer plasmid DNA via eletroporation will greatly facilitate the application of recombinant DNA methodology and transposon technology to Gram-positive bacteria for cloning and analysis of significant genes.
通过电穿孔将质粒DNA导入芽孢杆菌属、肠球菌属、乳杆菌属、乳球菌属、明串珠菌属、李斯特菌属、片球菌属、丙酸杆菌属和葡萄球菌属，作为感受态细胞或原生质体转化的替代方法。在这些受体中回收了含质粒的转化体，频率范围为每微克pGK12有10(1)至10(5)个转化体。评估了该方案的几个参数，包括DNA浓度、电压、平板接种方案和电穿孔缓冲液，以确定提高嗜酸乳杆菌转化频率的条件。使用优化条件，将以下质粒导入嗜酸乳杆菌：pAMB1、pC194、pGB354、pGKV1、pSA3、pTRK13、pTV1和pVA797。通过电穿孔转移质粒DNA的能力将极大地促进重组DNA方法和转座子技术在革兰氏阳性菌中的应用，用于重要基因的克隆和分析。
