Cloning of pediocin PA-1 and its immunity genes from Pediococcus acidilactici K7 using pAMJ shuttle vector into Lactococcus lactis MG1363

The matured pediocin PA-1 encoding gene, pedA and its
immunity counterpart, pedB from Pediococcus acidilactici K7
were cloned by PCR technique and ligated into the E. coli-lactic
shuttle, protein expression vector pAMJ2OO8. The recombinant
pAML45 was constructed and sequenced to confirm the intactness
of reading frame of the deduced pediocin PA-1 fusion protein.
Lactococcus lactis MG1363 was electroporated with the plasmids
and the transformation efficiency of ~ 2-3 × 103 μg-1 was
observed. This was further confirmed by plasmid DNA isolation
and analysis.