New method for electroporation of Lactobacillus species grown in high salt |
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Authors: | Maria Mercedes Palomino |
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Institution: | Universidad de Buenos Aires, FCEyN, Dpto. de Química Biológica, Bs As, Argentina |
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Abstract: | We here describe a new method for electroporation of Lactobacillus species, obligately homofermentative and facultatively heterofermentative, based on the cell-wall weakening resulting from growth in high-salt media. For L. casei, optimum transformation efficiency of up to 105 transformants per microgram of plasmid DNA was achieved following growth in the presence of 0.9 M NaCl. Plasmids of different sizes and replication origins were also similarly transformed. These competent cells could be used either directly or stored frozen, up to 1 month, for future use, with similar efficiency. This protocol was assayed with different Lactobacillus species: L. delbrueckii subsp. lactis, L. paracasei, L. plantarum and L. acidophilus, and it was found that they were transformed with similar efficiency. |
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Keywords: | Lactobacillus Electroporation High salt growth |
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