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Nucleic Acids Research, 1992, Vol. 20, No. 11 2705-2710
© 1992

MOLECULAR BIOLOGY

Structural and functional features of cis-acting sequences in the basic replicon of plasmid Collb-P9

Katsunori Tanaka, Hiroshi Sakai1, Yoichi Honda, Toshiaki Nakamura, Akihisa Higashi1 and Tohru Komano*

Laboratory of Biochemistry, Department of Agricultural Chemistry, Kyoto University, Kyoto 606 1Research and Development Center, DAIKEN Corporation, Okayama 702, Japan

*To whom correspondence should be addressed

Received March 20, 1992. Accepted April 9, 1992.

We have structurally and functionally analyzed the cis-elements essential for Collb-P9 plasmid DNA replication. The putative oriV region encompassed a region of 172 base pairs (bp) located 152 bp downstream of the repZ gene. A typical dnaA box found in this region proved nonessential for the DNA replication of Collb-P9. The ssi signal of Collb-P9 is a homologue of the G-sites of R1 and R100 plasmids. Deletion of the G-site led to 1.5-fold reduction of the copy number, suggesting that although this G-site is not essential, It is important for efficient Collb-P9 DNA replication. In addition, the Collb-P9 replicon is highly and extensively homologous with the P307 (RepFIC) replicon, and highly homologous with the R100 (RepFIIA) replicon around the G-site region. These facts imply a common ancestry from which the plasmids have evolved.


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