Open AccessDevelopment and application of molecular tools in the study of IncN‐related plasmids from lakewater sediments
Author(s) -
Osborn A.Mark,
Pickup Roger W.,
Saunders Jon R.
Publication year - 2000
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2000.tb09105.x
Subject(s) - plasmid , environmental chemistry , biology , environmental science , ecology , chemistry , genetics , gene
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.