Open AccessOkazaki pieces grow opposite to the replication fork direction during simian virus 40 DNA replication
Author(s) -
Gabriel Kaufmann,
Rachel Bar-Shavit,
Melvin L. DePamphilis
Publication year - 1978
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/5.7.2535
Subject(s) - okazaki fragments , biology , dna replication , dna , replication factor c , microbiology and biotechnology , control of chromosome duplication , dna ligases , dna polymerase ii , eukaryotic dna replication , primer (cosmetics) , genetics , polymerase chain reaction , gene , chemistry , reverse transcriptase , organic chemistry
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 Bg1(1)+Hpa1 simian virus 40 DNA restriction fragments immobilized on nitrocellulose 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.