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Journal of Bacteriology, July 2005, p. 4728-4738, Vol. 187, No. 14
0021-9193/05/$08.00+0     doi:10.1128/JB.187.14.4728-4738.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

The IncP-6 Plasmid Rms149 Consists of a Small Mobilizable Backbone with Multiple Large Insertions

Anthony S. Haines, Karen Jones, Martin Cheung, and Christopher M. Thomas*

School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom

Received 2 January 2005/ Accepted 20 April 2005

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.

* Corresponding author. Mailing address: School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom. Phone: 44 121 414 5903. Fax: 44 121 414 5925. E-mail: c.m.thomas{at}bham.ac.uk.

Journal of Bacteriology, July 2005, p. 4728-4738, Vol. 187, No. 14
0021-9193/05/$08.00+0     doi:10.1128/JB.187.14.4728-4738.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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