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Start Sites of Bidirectional DNA Synthesis at the Human Lamin B2 Origin

Science  17 Mar 2000:
Vol. 287, Issue 5460, pp. 2023-2026
DOI: 10.1126/science.287.5460.2023

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Abstract

The initiation sites of bidirectional synthesis at the DNA replication origin located at the 3′ end of the human lamin B2 gene were investigated. RNA-primed nascent DNA molecules were subjected to second-strand synthesis with appropriate primers, amplified by ligation-mediated polymerase chain reaction, and size fractionated. Evidence for precise start sites was obtained. Exploration of close to 1 kilobase, coupled to inhibition of Okazaki fragment synthesis, demonstrates that the leading strands initiate at precise nucleotides on either helix, overlapping by three base pairs, within the area bound to a protein complex possibly analogous to the prereplicative complex of yeast.

  • * These authors contributed equally to this work.

  • To whom correspondence should be addressed. E-mail: falaschi{at}icgeb.trieste.it

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