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J Virol. 1967 April; 1(2): 302-307
Copyright © 1967 American Society for Microbiology. All Rights Reserved.

Purification and Properties of Reovirus Ribonucleic Acid

¹ The Public Health Research Institute of The City of New York, Inc., New York, New York 10009

ABSTRACT

NaClO₄ was employed in a technique for the rapid extraction of reovirus ribonucleic acid (RNA). The extracted RNA, which was purified in a Cs₂SO₄ equilibrium density gradient, had a buoyant density of 1.61 g/cm³ 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.