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Nucleic Acids Research, 1978, Vol. 5, No. 7 2535-2546
© 1978

Articles

Okazaki pieces grow opposite to the replication fork direction during simian virus 40 DNA replication

Gabriel Kaufmann*,+, Rachel Bar-Shavit* and Melvin L. DePamphilis**

*The Biochemistry Department, The Weizmann Institute of Science Rehovot, Israel **Department of Biological Chemistry, Harvard Medical School Boston, MA 02115, USA

+To Whome all correspondence should be address

Received April 19, 1978. Simian virus 40 replicating DNA was pulse labeled with {alpha}-32p-dATP using an acellular DNA replication system. Nascent DNA chains of less than 200 nucleotides (Okazaki pieces) were then isolated from the denatured replicating DNA by electrosieving through a polyacrylamide gel column. The purified Okazaki pieces were hybridized to separated strands of Bg11+Hpa1 simian virus 40 DNA restriction fragments immobilized on nitrocellutose filters. only strands with polarity of the DNA replication fork direction hybridized with Okazaki pieces. Hence, Okazaki pieces in simian virus 40 are synthesized against the DNA replication fork direction.


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