Genetic transformation of Porphyromonas gingivalis by electroporation

Abstract

Porphyromonas gingivalis was transformed by electroporation using the DNA of plasmid pE5-2, or its derivative, pYT7. Prior to transformation, pE5-2 was transferred from Escherichia coli to P. gingivalis strains by conjugation (mobilization with R751), and the plasmid DNA was purified from the P. gingivalis transconjugants. Transformation occurred when the recipient strain and the donor strain from which the plasmid DNA was purified were homologous. If they were heterologous, transformation did not take place or did so at a very low frequency. This suggested that a restriction-modification system is present in P. gingivalis strains. Plasmid pYT7 was derived by removing an 8.0 kb AvaI fragment from pE5-2 that was purified from P. gingivalis cells. It has several single-cutting restriction sites such as EcoRI, AvaI and ClaI usable for gene cloning, though it was not stable enough in P. gingivalis cells, probably because the rep gene was derived from a relatively distant species, Bacteroides eggerthii.