Transfection of Actinomyces spp. by genomic DNA of bacteriophages from human dental plaque.
Bacteriophages that produced turbid or clear zones of lysis in strains of Actinomyces were isolated from 22 of 124 samples of fresh human dental plaque. All human and nonhuman strains of Actinomyces viscosus or Actinomyces naeslundii tested in this study were sensitive to infection by one or more of these phages. In contrast, none of the Actinomyces odontolyticus, Actinomyces israelii, or Actinomyces bovis strains tested were susceptible. Results of restriction endonuclease analyses indicated that the genomes of these phages consisted of double-stranded DNA molecules ranging in size between 16 and 60 kbp. Sequence homology under hybridization conditions of high stringency was observed among a few of the isolated phages. A lysogenized isolate of A. viscosus MG-1 was obtained following infection with a temperate phage, designated phi 225. Results of Southern blot analyses indicated that phi 225 replicated as a plasmid in the lysogenized strain. Genomic DNA from several lytic phages was used to establish conditions for transfection by electroporation of strains of Actinomyces spp. Efficiencies of DNA transfer ranged from 10(2) to 10(5) plaque-forming units per microgram of DNA were obtained under optimal transfection conditions. The results of these studies demonstrate that transfer of genetic information in Actinomyces spp. can be achieved by transfection.
Yeung
M K
MK
Department of Pediatric Dentistry, University of Texas Health Science Center at San Antonio 78284-7888, USA. yeung@uthscsa.edu
Kozelsky
C S
CS
D000190:Actinomyces / Q000235:genetics* / Q000302:isolation & purification / Q000821:virology*; D000818:Animals; D001435:Bacteriophages / Q000235:genetics* / Q000302:isolation & purification; D004279:DNA, Viral / Q000235:genetics*; D003773:Dental Plaque / Q000821:virology*; D005821:Genetic Techniques; D006801:Humans; D008242:Lysogeny / Q000235:genetics; D014162:Transfection
D004279:DNA, Viral