Identification of the origin of replication of the eukaryote Dictyostelium discoideum nuclear plasmid Ddp2

Andy C. M. Chang, Martin B. Slade, Keith L. Williams

Research output: Contribution to journalArticlepeer-review

27 Citations (Scopus)

Abstract

Ddp2 is a 5.8-kb, high-copy-number, nuclear plasmid found in the eukaryote Dictyostelium discoideum. We have identified two functional domains, a large open reading frame (Rep gene) and a 626-bp fragment containing an origin of replication (ori). The ori, when cloned into a shuttle vector, confers stable extrachromosomal replication in D. discoideum, provided that the Rep gene, which acts intrans, is integrated into the host genome. Ddp2 carries a 501-bp imperfect inverted repeat, and part of the ori overlaps with one of these repeats. The ori sequence contains two direct repeats of 49 bp comprising two 10-bp "TGTCATGACA" palindromes separated by a poly(T · A) sequence. Deletion of either 49-bp repeat abolished extrachromosomal replication.

Original languageEnglish
Pages (from-to)208-217
Number of pages10
JournalPlasmid
Volume24
Issue number3
DOIs
Publication statusPublished - 1990

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