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. 2004 Oct;8(5):385-91.
doi: 10.1007/s00792-004-0399-y. Epub 2004 Aug 5.

Identification and autonomous replication capability of a chromosomal replication origin from the archaeon Sulfolobus solfataricus

Patrizia Contursi  1 , Francesca M Pisani, Andrei Grigoriev, Raffaele Cannio, Simonetta Bartolucci, Mosè Rossi
Affiliations

Identification and autonomous replication capability of a chromosomal replication origin from the archaeon Sulfolobus solfataricus

Patrizia Contursi et al. Extremophiles. 2004 Oct.

Abstract

Here, we describe the identification of a chromosomal DNA replication origin (oriC) from the hyperthermophilic archaeon Sulfolobus solfataricus (subdomain of Crenarchaeota). By means of a cumulative GC-skew analysis of the Sulfolobus genome sequence, a candidate oriC was mapped within a 1.12-kb region located between the two divergently transcribed MCM- and cdc6-like genes. We demonstrated that plasmids containing the Sulfolobus oriC sequence and a hygromycin-resistance selectable marker were maintained in an episomal state in transformed S. solfataricus cells under selective pressure. The proposed location of the origin was confirmed by 2-D gel electrophoresis experiments. This is the first report on the functional cloning of a chromosomal oriC from an archaeon and represents an important step toward the reconstitution of an archaeal in vitro DNA replication system.

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