Treponema denticola is strongly implicated in the etiology of periodontal diseases. However, genetic transformation of this organism has not been reported. We now demonstrate a gene transfer system in T. denticola by electroporation using a broad host plasmid pKT210 as a shuttle vector. Plasmid extraction, Southern blot hybridization as well as the polymerase chain reaction indicated the presence of the plasmid in T. denticola transformants. The restriction patterns of plasmid pKT210 rescued from the T. denticola transformants in Escherichea coli suggested that some of the rescued plasmids were identical to the original pKT210, but some of them had been modified. This transformation system could be a potentially useful tool for genetic manipulation of oral spirochetes.