title: Feline calicivirus: purification of virus and extraction and characterisation of its ribonucleic acid.
abstract: Purification of feline calicivirus was achieved by cycles of differential centrifugation and two cycles of sucrose gradient centrifugation. Feline calicivirus grown in the presence of Actinomycin D and 3H-uridine-5, sediments in 15% to 45% sucrose gradients and forms a peak of radioactivity which corresponds with the peak of infectivity. Ribonucleic acid (RNA) extracted from the peak radioactive fractions taken from the sucrose gradient sedimented as a single peak ahead of the 28S peak of cellular RNA. It was sensitive to ribonuclease and was presumed to be single stranded feline calicivirus RNA with sedimentation of 32S-35S. A single peak of radioactivity at 35S was extracted from purified virus by heating at 60 degrees for two minutes in 1% sodium dodecyl sulphate (SDS), or by heating at 37 degrees for 5 minutes at 1% SDS. Virus extracted at 37 degrees for 10 minutes in 1% SDS showed also a small peak at 16S and by 15 minutes at 37 degrees only a broad peak at 16S occurred. All peaks were susceptible to ribonuclease. A component sedimenting at 18S which was resistent to degradation by ribonuclease under the conditions outlined by Baltimore (4) and presumed to be double-stranded RNA was present in kitten kidney cells infected with feline calicivirus.
mesh_terms: D002139:Caliciviridae / Q000032:analysis / Q000187:drug effects / Q000302:isolation & purification*; D010849:Picornaviridae / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification*; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology
chemical_list: D012367:RNA, Viral; D012967:Sodium Dodecyl Sulfate