Fate of free DNA and transformation of the oral bacterium Streptococcus gordonii DL1 by plasmid DNA in human saliva.
Competitive PCR was used to monitor the survival of a 520-bp DNA target sequence from a recombinant plasmid, pVACMC1, after admixture of the plasmid with freshly sampled human saliva. The fraction of the target remaining amplifiable ranged from 40 to 65% after 10 min of exposure to saliva samples from five subjects and from 6 to 25% after 60 min of exposure. pVACMC1 plasmid DNA that had been exposed to degradation by fresh saliva was capable of transforming naturally competent Streptococcus gordonii DL1 to erythromycin resistance, although transforming activity decreased rapidly, with a half-life of approximately 50 s. S. gordonii DL1 transformants were obtained in the presence of filter-sterilized saliva and a 1-microg/ml final concentration of pVACMC1 DNA. Addition of filter-sterilized saliva instead of heat-inactivated horse serum to S. gordonii DL1 cells induced competence, although with slightly lower efficiency. These findings indicate that DNA released from bacteria or food sources within the mouth has the potential to transform naturally competent oral bacteria. However, further investigations are needed to establish whether transformation of oral bacteria can occur at significant frequencies in vivo.
Mercer
D K
DK
Rowett Research Institute, Bucksburn, Aberdeen, Great Britain. d.mercer@rri.sari.ac.uk
Scott
K P
KP
Bruce-Johnson
W A
WA
Glover
L A
LA
Flint
H J
HJ
D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics*; D018014:Gene Transfer Techniques; D006801:Humans; D066298:In Vitro Techniques; D009055:Mouth / Q000382:microbiology; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012463:Saliva / Q000382:microbiology*; D013291:Streptococcus / Q000235:genetics*; D014170:Transformation, Genetic
D017931:DNA Primers; D004269:DNA, Bacterial