Optimization of Electroporation Processing of Lactobacilli

Abstract

Abstract: One of the genetic manipulation restriction factors in Lactobacillus strains is the efficient and reproducible transformation method. To optimize the conditions for electroporation of Lactobacillus plantarum B0080 cells, a shuttle vector for Escherichia coli and Bacillus subtilis, pHY300PLK, were used. Factors including growth stage, plasmid concentration and composition of the recovery medium were investigated. Response surface methodology was employed to optimize the growth medium and pulse strength. Results showed that the optimal values for concentration of glycine and of sucrose and the pulse strength are 5.13% (W/V), 0.54 mol/L and 7.41 kV/cm, respectively. Best electroporation efficiency is 3.6×104 CFU/μg DNA obtained after incubation in 0.3 mol/L sucrose containing MRS broth for another 2 h. Several strains of Lactobacillus, such as L.amylovorus B0112 and L.acidophilus B0068, are transformed with different kinds of plasmids with the optimized processing.

Key words: Lactobacillus plantarum, Lactobacillus amylovorus, pHY300PLK, electroporation

XIA Zi-Fang, SHI Gui-Yang, ZHANG Liang, CHEN Wei, WANG Zheng-Xiang. Optimization of Electroporation Processing of Lactobacilli[J]. FOOD SCIENCE, 2008, 29(2): 205-209.

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Optimization of Electroporation Processing of Lactobacilli

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