植物乳杆菌
电穿孔
质粒
大肠杆菌
转化(遗传学)
DNA
微生物学
细菌
生物
转化效率
乳酸菌
分子生物学
化学
乳酸
生物化学
基因
遗传学
农杆菌
作者
M. Teresa Alegre,M. Carmen RodrÃguez,Juan M. Mesas
标识
DOI:10.1016/j.femsle.2004.10.006
摘要
An improved method for the electrotransformation of Lactobacillus plantarum CECT 220 (ATCC 8014) with plasmid DNA isolated from Escherichia coli is described. The two main modifications with respect to existing methods are: (i) isolation of plasmid DNA from E. coli JM110 grown in minimal medium and (ii) in vitro modification of the DNA by cell-free extracts of the host L. plantarum. Optimal electrotransformation was obtained with exponentially growing cells of L. plantarum concentrated to 6x10(9) cfu ml-1, with electric pulses of 13 kV cm-1 in cuvettes with 1 mm inter-electrode distance. We consider that this method constitutes a useful tool for routine manipulation of L. plantarum, and can probably be extended to other lactic acid bacteria.
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