Development and application of molecular tools in the study of IncN-related plasmids from lakewater sediments.
Homology to IncN, P, Q and W inc regions was investigated amongst 114 Hg(2+)-resistant or antibiotic-resistant bacteria isolated from lakewater sediments. No hybridisation signals were found with Inc P, Q and W probes, and only one plasmid, pLV1402, hybridised to the IncN probe. PCR primers designed to conserved regions in the replicon of the IncN plasmid pCU1 and the related beta replicon from pGSH500 were used to amplify a 978-bp fragment from pLV1402, with sequence analysis showing a close relationship (99.2% identity) between their replication genes. A 387-bp region from the pLV1402 rep gene was used to re-screen the isolates and identified another related plasmid, pLV1403. A 3.7-kb probe containing the alpha replicon from pGSH500 hybridised to both pLV1402 and pLV1403, suggesting that both are multi-replicon plasmids. The PCR primers and probes described will be useful in future studies of plasmid diversity.
Osborn
A M
AM
School of Biological Sciences, Life Science Building, The University of Liverpool, P.O. Box 147, Liverpool, UK. osborn@essex.ac.uk
Pickup
R W
RW
Saunders
J R
JR
D001419:Bacteria / Q000235:genetics* / Q000302:isolation & purification*; D003001:Cloning, Molecular / Q000379:methods; D017124:Conserved Sequence; D017931:DNA Primers; D004352:Drug Resistance, Microbial / Q000235:genetics*; D004926:Escherichia coli; D005618:Fresh Water / Q000382:microbiology*; D005821:Genetic Techniques; D019015:Geologic Sediments / Q000382:microbiology*; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods; D012093:Replicon
D017931:DNA Primers