Mini-Tn7 transposons for site-specific tagging of bacteria with fluorescent proteins.
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, P(A1/04/03), 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.
Lambertsen
Lotte
L
Molecular Microbial Ecology Group, Centre for Biomedical Microbiology, BioCentrum-DTU, Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark. lol@biocentrum.dtu.dk
Sternberg
Claus
C
Molin
Søren
S
D019677:Artificial Gene Fusion; D015139:Blotting, Southern; D002876:Chromosomes, Bacterial; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D017930:Genes, Reporter; D007763:Lac Operon; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D016958:Pseudomonas putida / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D013194:Staining and Labeling; D012407:rRNA Operon
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D008164:Luminescent Proteins; D011994:Recombinant Proteins