title: Identification of the replication region of a 111-kb circular plasmid from Rhodococcus opacus B-4 by λ Red recombination-based deletion analysis.
abstract: The replication region of the 111-kb circular plasmid pKNR from Rhodococcus opacus B-4 was identified. A PCR-based deletion analysis using the λ Red recombination technique followed by restriction digestion and PCR-amplification analyses revealed that a 2.5-kb fragment covering one putative open reading frame (ORF) was involved in the replication of pKNR. The product of this ORF showed significant similarity to a functionally unknown protein encoded in the replication region of the 70-kb circular plasmid of Clavibacter michiganensis and to ones in other bacterial large circular plasmids. These observations suggest that the product of the identified ORF and its orthologs can serve as novel replication proteins for large circular bacterial plasmids.
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan. honda@bio.eng.osaka-u.ac.jp
authors.forename: Kohsuke
authors.initials: K
authors.lastname: Honda
authors.forename: Makoto
authors.initials: M
authors.lastname: Imura
authors.forename: Kenji
authors.initials: K
authors.lastname: Okano
authors.forename: Takeshi
authors.initials: T
authors.lastname: Omasa
authors.forename: Junichi
authors.initials: J
authors.lastname: Kato
authors.forename: Hisao
authors.initials: H
authors.lastname: Ohtake
mesh_terms: D001483:Base Sequence; D004261:DNA Replication; D004269:DNA, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D057927:Hydrophobic and Hydrophilic Interactions; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010957:Plasmids / Q000737:chemistry / Q000235:genetics*; D011995:Recombination, Genetic; D015183:Restriction Mapping; D012240:Rhodococcus / Q000235:genetics*; D017384:Sequence Deletion
chemical_list: D004269:DNA, Bacterial