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Home → Publications → American Chestnut Proceedings → 1982 → Isolation and Partial Characterization of a Virus-like Particle and it's Genome Associated With Endothia Parasitica Strain 43

Isolation and Partial Characterization of a Virus-like Particle and it's Genome Associated With Endothia Parasitica Strain 43

ABSTRACT.--A virus-like particle (VLP) infecting Endothia parasitica strain 43 has been partially purified by polyethylene glycol (PEG) precipitation, differential centrifugation and isopycnic density gradient centrifugation in cesium chloride (CsCl) and cesium sulphate (Cs2SO4). The buoyant density of the particle was determined to be 1.207 and 1.177 g/cc respectively. A sedimentation (S) value of 164 was calculated for the particle. Examined under the electron microscope, the VLP's were pleiomorphic in morphology with a diameter of 100 nm. The particles possessed an ultraviolet absorption profile characteristic of nucleoproteins. The viral genome was extracted, partially purified and found to consist of two segments of double-stranded ribonucleic acid (dsRNA). The molecular weights of the two components were resolved by polyacrylamide gel electrophoresis to be 6.76 and 6.02 x 106 daltons. The dsRNA possessed an S value of 34, a buoyant density of 1.597g/cc in Cs2SO4, and upon thermal denaturation a 30 percent shift in absorbance was observed with a Tm value of 89.3 C. Partially purified preparations from a virulent strain lacked the VLP's and any detectable dsRNA.


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Author(s): Richard A. Chmelo, Walter J. Kaczmarczyk

Publication: American Chestnut Proceedings - 1982