Purification of herpesvirus saimiri and properties of the viral DNA

Conditions for growth, concentration, and purification of Herpesvirus saimiri were determined. Optimal yields of infectious Herpesvirus saimiri (HVS) were obtained from infected owl monkey kidney (OMK) cells grown at 32.5 degrees C in medium containing 10 per cent fetal calf serum. Forth-five percent of the initial infectious HVS was recovered after an 18-fold concentration using 8 per cent polyethylene glycol 6000 in the presence of 0.5 M NaCl. Polyethylene glycol concentrated HVS was purified in an isopycnic-linear Renografin gradient (1.0-1.3 g/cm3. Ninety-six percent of the infectivity was recovered in a single 1.16 g/cm3 density region. DNA extracted from purified HVS was resolved into two distinct density classes by CsCl equilibrium centrifugation (1.727 and 1.709 g/cm3). DNase treated HVA virions yield four DNA species with densities of 1.727, 1.718, 1.712, and 1.706 g/cm3 in CsCl centrifugation.