Affiliation:
1. Göteborg University, Department of Cell and Molecular Biology, Microbiology, SE-405 30 Göteborg, Sweden
Abstract
ABSTRACT
Gene transfer of the conjugative plasmid pBF1 from
Pseudomonas putida
to indigenous bacteria in seawater was investigated with a detection system for gene transfer based on the green fluorescent protein (GFP) (C. Dahlberg et al., Mol. Biol. Evol. 15:385–390, 1998). pBF1 was tagged with the
gfp
gene controlled by a
lac
promoter which is down regulated in the donor cell by a chromosomal repressor (
lacI
q
). The plasmid donor cells (
Pseudomonas putida
KT2442) subsequently do not express
gfp
. Transfer to recipient strains lacking the repressor results in expression of
gfp
. The transconjugant can subsequently be detected by epifluorescence microscopy on a single-cell level. By using this method, transfer of pBF1::
gfp
and expression of the
gfp
gene were first shown to occur during nutrient-limiting conditions to several defined recipient bacteria in artificial seawater. Second, we measured transfer of pBF1 from
P. putida
to the marine bacterial community directly in seawater samples, on a single-cell level, without limiting the detection of gene transfer to the culturable fraction of bacteria. Plasmid transfer was detected on surfaces and in bulk seawater. Seawater bacteria with different morphologies were shown to receive the plasmid. Gene transfer frequencies of 2.3 × 10
−6
to 2.2 × 10
−4
transconjugants per recipient were recorded after 3 days of incubation.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Cited by
110 articles.
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