Abstract
When cells infected with Kilham rat virus (KRV) were incubated in 4 M guanidine-HCl, the single-stranded DNA was completely released from virions of KRV, but double-stranded DNA was not denatured. The three forms of viral DNA released had properties, respectively, of double-stranded replicative-form DNA, single-stranded progeny DNA, and an intermediate DNA species. Sucrose gradients containing 4 M guanidine-HCl were used to isolate these intracellular DNA forms with complete separation from high molecular weight cellular DNA. After a short pulse, radioactivity in low molecular weight DNA was found almost exclusively in molecules having sedimentation properties of viral replicative-form DNA. During chase incubation, this radioactivity progressively shifted to the position of single-stranded progeny DNA.
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