Mini-Tn7 transposons for site-specific tagging of bacteria with fluorescent proteins

Summary The mini‐Tn7 transposon system is a convenient tool for site‐specific tagging of bacteria in which the tagging DNA is inserted at a unique and neutral chromosomal site. We have expanded the panel of mini‐Tn7 delivery plasmids expressing different fluorescent proteins (stable and unstable) fr...

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Bibliographic Details
Published in:Environmental microbiology 2004-07, Vol.6 (7), p.726-732
Main Authors: Lambertsen, Lotte, Sternberg, Claus, Molin, Søren
Format: Article
Language:English
Subjects:
Artificial Gene Fusion
Blotting, Southern
Chromosomes, Bacterial
DNA Transposable Elements
DNA, Bacterial - analysis
DNA, Bacterial - isolation & purification
Escherichia coli - genetics
Genes, Reporter
Lac Operon
Luminescent Proteins - genetics
Luminescent Proteins - metabolism
Plasmids
Polymerase Chain Reaction
Promoter Regions, Genetic
Pseudomonas putida
Pseudomonas putida - genetics
Pseudomonas putida - growth & development
Pseudomonas putida - metabolism
Recombinant Proteins - genetics
Recombinant Proteins - metabolism
Recombination, Genetic
rRNA Operon
Staining and Labeling
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Summary:Summary The mini‐Tn7 transposon system is a convenient tool for site‐specific tagging of bacteria in which the tagging DNA is inserted at a unique and neutral chromosomal site. We have expanded the panel of mini‐Tn7 delivery plasmids expressing different fluorescent proteins (stable and unstable) from the Escherichia coli lac derived promoter, PA1/04/03, or from the growth‐rate‐dependent Escherichia coli promoter PrrnB P1. The mini‐Tn7 transposons were inserted and tested in the soil bacterium, Pseudomonas putida KT2440. Successful and site‐specific tagging was verified by Southern blots as well as by PCR. Furthermore, the effect of fluorescent protein expression on the cellular growth rate was tested by growth competition assays.
ISSN:1462-2912
1462-2920
DOI:10.1111/j.1462-2920.2004.00605.x