Plasmid transfer detection in soil using the inducible [lambda]Pr system fused to eukaryotic luciferase genes

We report a model system for plasmid transfer analysis using the regulated lambda phage right promoter, λPr, fused to luc and lucOR as repoter genes. We have demonstrated that the systems cI857-λPr::luc and cI857-λPr::lucOR are temperature-inducible in Escherichia coli but not in other Gram-negative...

Full description

Saved in:
Bibliographic Details
Published in:Microbial ecology 2001-05, Vol.41 (4), p.352
Main Authors: Palomares, A J, Vázquez, M E, Rodríguez-llorente, I D, Dary, M, Caviedes, M A
Format: Article
Language:English
Subjects:
Antibiotics
Bacteria
E coli
Microbiology
Microcosms
Microorganisms
Plasmids
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:We report a model system for plasmid transfer analysis using the regulated lambda phage right promoter, λPr, fused to luc and lucOR as repoter genes. We have demonstrated that the systems cI857-λPr::luc and cI857-λPr::lucOR are temperature-inducible in Escherichia coli but not in other Gram-negative bacteria analyzed, enabling detection of luminescence when plasmids were mobilized from E. coli to those Gram-negative backgrounds. Using light for the detection, we have observed plasmid transfer from E. coli harboring RK2 and R388 derived plasmids to Pseudomonas putida KT2440 (co-introduced with donors) and to indigenous microorganisms, in vitro and in nonsterile soil microcosms. The importance of nutrients for an efficient plasmid transfer in nonsterile soil microcosms has been confirmed. When plasmid transfer experiments were carried out into nonsterile soil microcosms, significant populations of indigenous transconjugants arose. This system provides efficient marker genes and avoids the use of antibiotics for the selection of transconjugants.[PUBLICATION ABSTRACT]
ISSN:0095-3628
1432-184X
DOI:10.1007/s002480000093