Exact(8)
Inoculated medium showing an opaque halo around the zone of growth was considered positive.
A central observation in this study were the characteristic expanding opaque halo zones, which surrounded all φ15 plaques.
When these particles were dropped onto a bacterial lawn, an opaque halo zone appeared after overnight incubation at 30°C.
The most distinct plaque feature is the opaque halo zone with an increasing diameter over the course of time, surrounding a plaque with constant diameter (Figure 1).
Phage φ15 forms plaques surrounded by growing opaque halo zones, indicative for EPS degradation, on seven out of 53 P. putida strains.
An opaque halo zone surrounding the plaque was observed for each of the seven strains (PpG1, PpN, PpN3, RD6PR1, RD5PR2, RD8PR2, RD8PR3) out of the collection of 53 P. putida strains sensitive for φ15 infection.
Dropping 10 µl of this solution in parallel with a 108 pfu phage suspension on a bacterial PpG1 lawn confirmed the presence of the EPS degrading activity within the tail spike protein of φ15, as the recombinantly purified tail spikes formed identical looking opaque halo zones as those found around the lytic zone caused by phage infection.
ivanovii colonies (blue-green colonies with opaque halo).
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