A mycovirus has been purified from mycelia of Penicillium chrysogenum by isopycnic centrifugation in sucrose and in CsCl. Viral particles band with a buoyant density of 1.20 in sucrose and 1.38 in CsCl. Particles have icosohedral symmetry, are 35 nm in diameter, and have an absorption profile characteristic of nucleoprotein. One enzymatic activity, RNA polymerase, is associated with the purified mycophage. Nucleic acid extracted from purified virus has a buoyant density in CS2SO4 of 1.61, a molar extinction coefficient of εp (258 nm) of 7200, a s20, w of 13.0, and a pattern of circular dichroism characteristic of double-helical ribonucleic acid. Molecules of this double-stranded ribonucleic acid (dsRNA), examined by electron microscopy, have a mean contour length of 0.86 μm which corresponds to a molecular weight of about 2.0 × 106 daltons. This dsRNA is resolved further by acrylamide gel electrophoresis into three closely spaced bands. Thermal denaturation of the viral dsRNA is dependent on ionic strength and gives a linear relationship with the negative logarithm of the sodium ion concentration.