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Nucleic Acids Research, 1990, Vol. 18, No. 18 5515-5519
© 1990

MOLECULAR BIOLOGY

Autonomous replication sequences in an extrachromosomal element of a pathogenic Entamoeba histolytica

J. Grodberg+, N. Salazar, R. Oren and D. Mirelman*

Department of Biophysics and Center for Molecular Biology of Tropical Diseases, Weizmann Institute of Science Rehovot 76100, Israel

* To whom correspondence should be addressed

Received May 18, 1990. Revised August 29, 1990. Accepted August 29, 1990.

Entamoeba histolytica possesses a 24.5 kilobase plasmid-like molecule which encodes for the organism's ribosomal RNAs. Sequence analysis of this extrachromosomal element revealed the presence of AT rich sequences which show homology to the origin of replication of other lower eucaryotes. An 802 bp fragment containining these sequences was cloned into a yeast shuttle vector lacking the origin of replication and the construct tested for its ability to replicate autonomously in yeast. Mitotic stability tests as well as evidence for plasmid maintenance indicate that the transformed cells contained self-replicating episomes and not stably integrated molecules. The nucleotide sequence of this ARS-containing fragment is presented.

+Present address: Department of Cellular aad Molecular Physiology, Harvard Medical School, 25 Shattuck St., Boston, MA 02115, USA


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