Purification of plum pox (sharka) virus with the use of Triton X-100

Abstract

Plum pox virus was purified by adding up to 5% non-ionic detergent Triton X-100 to extracts clarified by low-speed centrifugation. After stirring for 1/2 h, the suspensions were subjected to 2 cycles of differential centrifugation followed by sucrose density-gradient centrifugation. Purity of the product was confirmed by electron microscopy and equilibrium density-gradient centrifugation in CsCl. The virus sedimented in the analytical ultracentrifuge as a single peak with a sedimentation coefficient of about 170 S at infinite dilution. Virus so purified showed an absorption spectrum with a minimum at 247 nm and a maximum at 263 nm. The modal length of the virus particles in purified preparations was 764 nm. Antiserum prepared had a specific titer of 4096.