New method for electroporation of Lactobacillus species grown high salt

PALOMINO MARÍA MERCEDES; ALLIEVI MARIANA C; PRADO ACOSTA MARIANO; SANCHEZ RIVAS CARMEN; RUZAL SANDRA M.

JOURNAL OF MICROBIOLOGICAL METHODS

ELSEVIER SCIENCE BV

Año: 2010 vol. 83 p. 164 - 167

0167-7012

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 ef.ciency 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 ef.ciency. 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 ef.ciency.