In vitro reassembly of infectious polyoma virions.

AUTOR(ES)

Brady, J N

RESUMO

Initial experiments in our laboratory have successfully reassembled infectious polyoma virions from dissociated virion products. Virions treated with ethyleneglycol-bis-N,N'-tetraacetic acid and the reducing agent beta-mercaptoethanol at pH 7.5 were dissociated to a 48S DNA-protein complex and capsomere subunits. The virion dissociation products were not infectious by plaque assay and lacked hemagglutination activity. These virion dissociation products were reassembled to intact virions by overnight dialysis against a reassembly buffer containing CaCl2, dimethyl sulfoxide, and Triton X-100 in phosphate-buffered saline at pH 7.4. The biophysical characteristics of the reassembled virions were identical to those of untreated virions in that the reassembled virions had a sedimentation value of 240S in sucrose gradients and a buoyant density of 1.315 g/cm3 in CsCl isopycnic gradients. The reassembled virions were intact as determined by electron microscopy and were found to be 60% resistant to DNase I treatment. Biologically, the reassembled purified virions were found to partially regain both hemagglutinating activity and plaque-forming ability.

Documentos Relacionados

• Generation of capsids from unstable polyoma virions.
• Immunological reactivity of antisera to sodium dodecyl sulfate-derived polypeptides of polyoma virions.
• In Vitro Reassembly of Shell-Like Particles from Disrupted Polyoma Virus
• Dissociation of polyoma virus by the chelation of calcium ions found associated with purified virions.
• Replicative intermediates of hepatitis B virus in HepG2 cells that produce infectious virions.