Detection of phages infecting Bacteroides fragilis HSP40 using a specific DNA probe.
Nine bacteriophage isolates of Bacteroides fragilis, obtained from urban sewage and pig faeces samples using four different host strains (HSP40, RYC4023, RYC2056 and RYC3318), were compared on the basis of morphology, host range, DNA restriction patterns, DNA hybridisation and protein composition. All the phages are siphovirus and, as judged from cleavage by restriction endonucleases, their genome is composed of double-stranded DNA of similar size ( approximately 51-kb). Host range analysis differentiated two types of phages: (1) phages that clearly infect B. fragilis strains HSP40 (B40-2, B23-1, B23-2, B23-3 and B23-4, of which B40-8 is the phage type); and (2) the group of bacteriophages that were not infectious for HSP40 (B56-1, B56-2 and B18-1). Similarity in DNA restriction patterns and protein characteristics was found in the HSP40 infectious phages. Anti-B40-8 serum recognised only the proteins of the phages of this type. Although all phages showed similar major protein sizes, minor specific bands were detected. Bacteriophages B56-1, B56-2 and B18-1 showed heterogeneity in their DNA restriction profiles although some degree of DNA-DNA homology between all genomes was observed. Southern blot analysis with phage B40-8 DNA based probes identified a 1.5-kb DNA region homologous for all HSP40 phage isolates, but absent in the genome of the other phage isolates that did not infect this bacterial strain. The homologous region was used as a specific probe to specifically detect B. fragilis HSP40 phages.
Puig
M
M
Department of Microbiology, Faculty of Biology, University of Barcelona, Avgda. Diagonal, 645, 08028-Barcelona, Spain. rosina@bio.ub.es
Jofre
J
J
Girones
R
R
D000818:Animals; D001435:Bacteriophages / Q000235:genetics / Q000302:isolation & purification* / Q000648:ultrastructure; D001441:Bacteroides fragilis / Q000821:virology*; D015139:Blotting, Southern; D015153:Blotting, Western; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis; D015247:Deoxyribonuclease HindIII; D004591:Electrophoresis, Polyacrylamide Gel; D005243:Feces / Q000821:virology; D016679:Genome, Viral; D008854:Microscopy, Electron; D009693:Nucleic Acid Hybridization / Q000379:methods; D015183:Restriction Mapping; D012680:Sensitivity and Specificity; D012722:Sewage / Q000821:virology; D017906:Siphoviridae / Q000235:genetics / Q000302:isolation & purification* / Q000648:ultrastructure; D013552:Swine / Q000821:virology
D015342:DNA Probes; D004279:DNA, Viral; D012722:Sewage; D015247:Deoxyribonuclease HindIII