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Science 17 March 2000:
Vol. 287. no. 5460, pp. 2023 - 2026
DOI: 10.1126/science.287.5460.2023
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Reports

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

Gulnara Abdurashidova, 1* Marta Deganuto, 1* Raffaella Klima, 1 Silvano Riva, 2 Giuseppe Biamonti, 2 Mauro Giacca, 3 Arturo Falaschi 1dagger

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.

1 Molecular Biology and
3 Molecular Medicine Units, International Centre for Genetic Engineering and Biotechnology, Padriciano 99, 34012 Trieste, Italy.
2 Istituto di Genetica Biochimica ed Evoluzionistica of the Consiglio Nazionale delle Ricerche, Via Abbiategrasso 207, 27100 Pavia, Italy.
*   These authors contributed equally to this work.

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

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