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Updated: Aug 17, 2026

Site-specific Bacterial Chromosome Engineering: ΦC31 Integrase Mediated Cassette Exchange (IMCE)
Published on: March 16, 2012
The IncP-6 plasmid Rms149 consists of a small mobilizable backbone with multiple large insertions
Anthony S Haines1, Karen Jones, Martin Cheung
1School of Biosciences, University of Birmingham, Edgbaston, UK.
Abstract:
Plasmid Rms149, the archetype of Pseudomonas plasmid incompatibility group IncP-6, was identified in Pseudomonas aeruginosa as an agent conferring resistance to streptomycin, sulfanilamide, gentamicin, and carbenicillin in 1975. It has been classed as a broad-host-range plasmid due to its ability to replicate in both Escherichia coli (where it is designated IncG) and Pseudomonas species, although both species are gamma-proteobacteria. To provide reference information on this Inc group, we have determined the complete sequence of Rms149 and found that, although the genome comprises 57,121 bp, it is essentially a small mobilizable plasmid carrying multiple mobile elements, which make up 79% (>45 kb) of its genome. A replicon has been identified which encodes a single polypeptide with moderate identity to other replication proteins. The region encoding this protein can replicate in Pseudomonas putida and E. coli. This sequence is directly downstream of a putative partitioning region highly similar to that of pRA2. A functional IncQ-type mobilization region is also present. Thus, the backbone appears to be a novel combination of modules already identified in other plasmid systems. Analysis of the segments that fall outside this core of stable inheritance and transfer functions show that this plasmid has been subject to multiple insertion events and that the plasmid appears to carry a considerable load of DNA that no longer should be phenotypically advantageous. The plasmid therefore functions not just as a vehicle for spread of selective traits but also as a store for DNA that is not currently under selection.
Insights
The Rms149 plasmid, a key IncP-6 group member, was sequenced, revealing a mobilizable plasmid with extensive mobile elements. Its novel backbone combines known modules, functioning as a vehicle for trait spread and a DNA storage system.
Area of Science:
- Microbiology
- Genetics
- Molecular Biology
Background:
- Plasmid Rms149, identified in 1975, belongs to the Pseudomonas plasmid incompatibility group IncP-6.
- It confers resistance to multiple antibiotics, including streptomycin, sulfanilamide, gentamicin, and carbenicillin.
- Rms149 is a broad-host-range plasmid, capable of replicating in both Pseudomonas species and Escherichia coli.
Purpose of the Study:
- To provide comprehensive reference information on the IncP-6 plasmid group.
- To determine the complete genome sequence of Rms149.
- To analyze the genetic structure and evolutionary implications of Rms149.
Main Methods:
- Whole-genome sequencing of Rms149.
- Bioinformatic analysis to identify functional regions and mobile elements.
- Comparative genomics to analyze plasmid backbone and insertion events.
Main Results:
- The Rms149 genome is 57,121 bp, with mobile elements comprising 79% of its sequence.
- A novel replicon was identified, enabling replication in Pseudomonas putida and E. coli.
- The plasmid backbone is a unique combination of known modules, with evidence of multiple insertion events.
Conclusions:
- Rms149 represents a novel combination of plasmid modules, functioning as a mobilizable plasmid.
- It serves as both a vehicle for spreading selective traits and a reservoir for non-selected DNA.
- The extensive mobile elements suggest a complex evolutionary history and potential for further genetic exchange.
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