Abstract
Abstract Fourteen to eighteen hours after low multiplicity infection of monolayer cultures of KB cells with adenovirus type 2 (Ad. 2), viral DNA was labeled with short pulses of tritiated thymidine and selectively extracted using sodium deoxycholate. After it had been purified, viral DNA was fractionated into partly single-stranded (or replicating) molecules and double-stranded (or completed) molecules on columns of benzolyatednaphthoylated DEAE (BND) cellulose. Completed molecules were cleaved with endonuclease R 1 from E. coli prior to electrophoresis through agarose-acrylamide slab gels. In agreement with previous results ( Winnacker, 1974 ; Schilling et al. , 1975 ), tritium recovery in the resulting fragments indicated that termination of replication occurred at the molecular ends of the genome. Replicating molecules were exposed to a single-strand specific nuclease from Neurospora crassa before being treated with endonuclease R 1 . In this instance, maximal tritium recovery was obvserved for the R 1 fragments corresponding to the center of the adenovirus DNA molecule. These results suggest that replication originates at the center of the molecule and proceeds in the direction of both ends.
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