Origin and Isolation

Phage LC16/VCM1 is a lytic myophage of Microcystis aeruginosa UTEX strain 2386 (isolated by W.W. Carmichael from Little Rideau Lake, Ontario, Canada).  This phage was isolated from the Copco Reservoir, which is a part of the Klamath River System in Northern California, by the following method:

1) 250mL of water from the surface of the Copco Reservoir was filtered with a 0.2 micron filter

2) The filtrate was enriched with BG-11 nutrients, inoculated with the target bacterial host strain: M. aeruginosa UTEX strain 2386, and incubated for approximately one week

3) Chloroform was added to the culture and then the aqaueous phase was filtered with a 0.2 micron filter

4) The filtrate was concentrated by ultracentrifugation and resuspended in SM buffer

5) The concentrated phage suspension was mixed with the target bacterial host strain in BG-11 top agar and plated on BG-11 agar plates

6) Ten individual plaques were selected from the top agar plate, picked from the plate, and resuspended in SM buffer for further phage propogation and characterization

Characterization

A 40,000x TEM micrograph of phage LC16/VCM1 indicates that it is a myophage with an icosahedral head and tapered tail with a baseplate.

PFGE

PFGE reveals that phage LC16/VCM1 posesses a genome of approximately
180kb. When 10 individual plaques from the same plate were isolated
(see the Origin and Isolation section above) and run
side-by-side in the PFGE apparatus, they all displayed the same 180kb
genome size. In addition, restriction digestion with either EcoRI or
BamHI gave identical restriction fragment length polymorphisms for each
plaque isolate.