Purification and properties of reovirus ribonucleic acid.
NaClO(4) was employed in a technique for the rapid extraction of reovirus ribonucleic acid (RNA). The extracted RNA, which was purified in a Cs(2)SO(4) equilibrium density gradient, had a buoyant density of 1.61 g/cm(3) and a sedimentation coefficient of 15S in a 7 to 20% sucrose gradient. It was 90% resistant to ribonuclease in a solution of high ionic strength (0.1 m NaCl). The sensitivity of reovirus RNA to ribonuclease increased with decreasing ionic strength. The thermal denaturation transition of the RNA began at 78 C and was complete at 85 C. The T(m) of the transition was 81 C in 0.01 m tris(hydroxymethyl)aminomethane buffer (pH 7.2) containing 0.001 m ethylenediaminetetraacetate. Thermal denaturation of reovirus RNA resulted in the formation of three ribonuclease-sensitive fractions. Denaturation at 25 C in the presence of dimethyl sulfoxide resulted in the formation of two ribonuclease-sensitive fractions.
D002499:Centrifugation, Density Gradient; D004121:Dimethyl Sulfoxide; D006358:Hot Temperature; D010472:Perchlorates; D012367:RNA, Viral / Q000302:isolation & purification*; D012087:Reoviridae; D012260:Ribonucleases; D012965:Sodium Chloride
D010472:Perchlorates; D012367:RNA, Viral; D012965:Sodium Chloride; D012260:Ribonucleases; D004121:Dimethyl Sulfoxide