Fate of free DNA and transformation of the oral bacterium Streptococcus gordonii DL1 by plasmid DNA in human saliva

D K Mercer, K P Scott, W A Bruce-Johnson, L A Glover, Harry James Flint

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

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-mu g/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.

Original languageEnglish
Pages (from-to)610
Number of pages5
JournalApplied and Environmental Microbiology
Volume65
Publication statusPublished - 1999

Keywords

  • NATURAL GENETIC-TRANSFORMATION
  • MOLECULAR-CLONING
  • BACILLUS-SUBTILIS
  • ESCHERICHIA-COLI
  • SANGUIS
  • MUTANS
  • TRACT
  • MICE
  • PLAQUES
  • LACTIS

Cite this

Fate of free DNA and transformation of the oral bacterium Streptococcus gordonii DL1 by plasmid DNA in human saliva. / Mercer, D K ; Scott, K P ; Bruce-Johnson, W A ; Glover, L A ; Flint, Harry James.

In: Applied and Environmental Microbiology, Vol. 65, 1999, p. 610.

Research output: Contribution to journalArticle

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abstract = "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-mu g/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.",
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AU - Scott, K P

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AU - Flint, Harry James

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N2 - 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-mu g/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.

AB - 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-mu g/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.

KW - NATURAL GENETIC-TRANSFORMATION

KW - MOLECULAR-CLONING

KW - BACILLUS-SUBTILIS

KW - ESCHERICHIA-COLI

KW - SANGUIS

KW - MUTANS

KW - TRACT

KW - MICE

KW - PLAQUES

KW - LACTIS

M3 - Article

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JO - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

SN - 0099-2240

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