Identification and autonomous replication capability of a chromosomal replication origin from the archaeonSulfolobus solfataricus.

Here, we describe the identification of a chromosomal DNA replication origin (oriC) from the hyperthermophilic archaeonSulfolobus solfataricus(subdomain of Crenarchaeota). By means of a cumulative GC-skew analysis of theSulfolobusgenome sequence, a candidateoriCwas mapped within a 1.12-kb region located between the two divergently transcribedMCM- andcdc6-like genes. We demonstrated that plasmids containing theSulfolobus oriCsequence and a hygromycin-resistance selectable marker were maintained in an episomal state in transformedS. solfataricuscells 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 chromosomaloriCfrom an archaeon and represents an important step toward the reconstitution of an archaeal in vitro DNA replication system. [ABSTRACT FROM AUTHOR]

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