The 20 bp, directly repeated DNA sequence of broad host range plasmid R1162 exerts incompatibility in vivo and inhibits R1162 DNA replication in vitro
- 1 July 1987
- journal article
- research article
- Published by Springer Nature in Molecular Genetics and Genomics
- Vol. 208 (3) , 390-397
- https://doi.org/10.1007/bf00328129
Abstract
The broad host range plasmid R1162 contains a directly repeated, 20 bp DNA sequence in the region of the plasmid required in cis for replication and maintenance. This sequence has been chemically synthesized and cloned, and shown to be sufficient for expression of plasmid incompatibility. The sequence also inhibits replication of R1162 DNA in a cell-free system. The strengths of both these effects are determined by the number of direct repeats (DRs) present, and are also affected to similar degrees by different mutations within the repeated sequence. Several of the mutations were tested for their effect in cis on plasmid maintenance in the cell, and one was found to cause an increase in plasmid copy number. The results suggest that the direct repeats exert incompatibility by inhibiting DNA replication, presumably because they are the binding sites for a limiting essential protein.Keywords
This publication has 43 references indexed in Scilit:
- Directly repeated, 20-bp sequence of plasmid R1162 DNA is required for replication, expression of incompatibility, and copy-number controlPlasmid, 1986
- Copy number of the broad host-range plasmid R1162 is determined by the amounts of essential plasmid-encoded proteinsJournal of Molecular Biology, 1985
- P1 plasmid replication: Replicon structureJournal of Molecular Biology, 1984
- Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesisGene, 1983
- A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragmentsGene, 1982
- A procedure for the large-scale isolation of highly purified plasmid DNA using alkaline extraction and binding to glass powderAnalytical Biochemistry, 1982
- Vertical dye-buoyant density gradients for rapid analysis and preparation of plasmid DNAAnalytical Biochemistry, 1981
- Nine unique repeating sequences in a region essential for replication and incompatibility of the mini-F plasmidGene, 1981
- Complete Nucleotide Sequence of the Escherichia coli Plasmid pBR322Published by Cold Spring Harbor Laboratory ,1979
- Analysis of bacteriophage Mu and λ-Mu hybrid DNAs by specific endonucleasesJournal of Molecular Biology, 1975