An improved method for the electrotransformation of lactic acid bacteria: A comparative survey

Genetic engineering of Geobacillus spp.

“…Quite efficient protocols have also been elaborated for several Bacillus and Lactobacillus strains (Landete et al, 2014;Vojcic et al, 2012). However, thermophilic Gram-positive bacteria are usually highly recalcitrant to genetic transformation.…”

Coproduction of colicin V and lactic acid bacteria bacteriocins in lactococci and enterococci strains of biotechnological interest

“…Preparation of competent cells and procedure of transformation of Lactococcus and Enterococcus were performed according to Landete et al . (). After 48–72 h of incubation, antibiotic‐resistant clones were screened for the presence of the insert of interest by PCR, using F‐pNZ8048/R‐Colicin.pNZ.…”

“…The PCR product was digested with the two restriction enzymes and ligated into the corresponding restriction sites of vector pT1NX vector digested with both of restriction endonucleases PstI and XbaI. The ligation mixture was used to transform L. lactis MG1363 by electroporation (Landete et al 2014).…”

“…The PCR product was digested with the two restriction enzymes and ligated into the corresponding restriction sites of vector pNZ:LR. The ligation mixture was used to transform L. lactis MG1363 by electroporation (Landete et al 2014). The resulting plasmid, named pNZ:LR-colV, was subsequently introduced into L. lactis MG1363 and transformants were checked by PCR with primers F-pNZ8048/R-Colicin.pNZ and sequencing of the inserted fragment.…”

“…Subsequently, plasmid pNZ:LR-colV was used to transform L. lactis ESI515, L. lactis TAB 26, L. lactis 639, L. lactis TAB 50 and Enterococcus faecium TAB 7 strains showed in Table 1. Preparation of competent cells and procedure of transformation of Lactococcus and Enterococcus were performed according to Landete et al (2014). After 48-72 h of incubation, antibiotic-resistant clones were screened for the presence of the insert of interest by PCR, using F-pNZ8048/R-Colicin.pNZ.…”