APPLICATION OF ELECTROPORATION FOR TRANSFER OF PLASMID DNA TO LACTOBACILLUS, LACTOCOCCUS, LEUCONOSTOC, LISTERIA, PEDIOCOCCUS, BACILLUS, STAPHYLOCOCCUS, ENTEROCOCCUS AND PROPIONIBACTERIUM
APPLICATION OF ELECTROPORATION FOR TRANSFER OF PLASMID DNA TO LACTOBACILLUS, LACTOCOCCUS, LEUCONOSTOC, LISTERIA, PEDIOCOCCUS, BACILLUS, STAPHYLOCOCCUS, ENTEROCOCCUS AND PROPIONIBACTERIUM
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DOI：
10.1111/j.1365-2958.1988.tb00072.x
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发表时间：
1988-09-01
影响因子：
3.6
通讯作者：
KLAENHAMMER, TR
中科院分区：
文献类型：
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作者：
LUCHANSKY, JB;MURIANA, PM;KLAENHAMMER, TR
用电穿孔的方法将质粒DNA导入芽孢杆菌、肠球菌、乳杆菌、乳杆菌、明串珠菌、李斯特菌、小球杆菌、丙酸杆菌和葡萄球菌，以替代感受态细胞或原生质体的转化。在这些受体中以pGK12的101到105个转化子·g-1的频率恢复了含有质粒的转化子。考察了DNA浓度、电镀工艺和电穿孔缓冲液等参数对嗜酸乳杆菌转化效率的影响。利用优化的条件，将pAMB1、pC194、pGB354、pGKV1、pSA3、pTRK13、pTV1和pVA797质粒导入嗜酸乳杆菌。通过电穿孔转移质粒DNA的能力将极大地促进重组DNA方法学和转座子技术在革兰氏阳性菌中的应用，以克隆和分析重要基因。
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 101 to 105 transformants .mu.g-1 of pGK12. Several parameters of the protocol, including DNA concentrations, 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 electroporation will greatly facilitate the application of recombinant DNA methodology and transposon technology to Gram-positive bacteria for cloning and analysis of significant genes.