Intercrossing of phage genomes in a phage cocktail and stable coexistence with Escherichia coli O157:H7 in anaerobic continuous culture.
The emergence of phage-resistant cells is the most serious problem for realizing phage therapy and is observed frequently if only one phage strain is used against a particular bacterium. By contrast, using multiple phages (phage cocktail) can delay or control the appearance of phage-resistant cells. Anaerobic continuous culturing of Escherichia coli O157:H7 and a cocktail of EP16, PP17, and SP22 phages were conducted. Comparison of the restriction fragment length polymorphism (RFLP) pattern of each phage genome showed a pattern different from wild type. Furthermore, the RFLP pattern of mutant phages consisted of fragments of PP17 and SP22 genome, suggesting both phages had infected the same host simultaneously (superinfection) and exchanged genomic DNA. Through observation of the binding of SYBR Gold-stained mutant phage to individual phage-resistant cells (RC), we found that clonal RC cultures were heterogeneous in their ability to bind mutant phage. The ratio of susceptibility was a few percent, which suggested that a minority of the RC population was susceptible to phage, and this heterogeneity contributes to the stable coexistence of RC and chimeric phages. The ratio of susceptible cells did not change appreciably from bacterial generation to generation.
Kunisaki
Hiroya
H
Department of Bioengineering, Tokyo Institute of Technology, Midori-ku, Yokohama 226-8501, Japan.
Tanji
Yasunori
Y
D000327:Adsorption; D000693:Anaerobiosis; D001287:Attachment Sites, Microbiological; D001433:Bacteriolysis; D003090:Coliphages / Q000235:genetics* / Q000254:growth & development* / Q000378:metabolism; D015169:Colony Count, Microbial; D004279:DNA, Viral / Q000235:genetics; D019453:Escherichia coli O157 / Q000235:genetics / Q000502:physiology* / Q000821:virology*; D016679:Genome, Viral; D008969:Molecular Sequence Data; D009154:Mutation; D012150:Polymorphism, Restriction Fragment Length
D004279:DNA, Viral