Back to Search
Start Over
Chromosomal gfp labelling of Pseudomonas aeruginosa using a mini-Tn7 transposon: application for studies of bacteria-host interactions.
- Source :
-
Canadian journal of microbiology [Can J Microbiol] 2008 Jan; Vol. 54 (1), pp. 48-57. - Publication Year :
- 2008
-
Abstract
- Analysis of bacterial interactions with host cells using multiple techniques is essential for studies on microbial pathogenesis and for the development of new antimicrobial therapies. Pseudomonas aeruginosa is an important opportunistic pathogen that can cause severe, often life-threatening pulmonary infections in individuals with impaired host defense mechanisms. Using a mini-Tn7 transposon delivery system, we have chromosomally labelled the strain P. aeruginosa PAK with a green fluorescent protein gene (gfp) and tested PAKgfp as a research tool for studies of bacteria-host interactions. We were able to reliably and rapidly measure the interactions of PAKgfp with A549 human lung epithelial cells by using flow cytometry, a fluorometric microplate reader-based assay, and fluorescence microscopy. With these analytical tools, we have demonstrated the adhesion of PAKgfp to the extracellular matrix protein fibronectin and the involvement of fibronectin in PAKgfp-A549 cell interactions. PAKgfp can be successfully used to explore the effects of various pharmacological compounds on P. aeruginosa - host cell interactions in both in vitro and in vivo systems, with potentially important medical applications.
- Subjects :
- Bacterial Adhesion
Cell Line
Epithelial Cells microbiology
Fibronectins metabolism
Flow Cytometry
Green Fluorescent Proteins genetics
Humans
Microscopy, Fluorescence
Mutagenesis, Insertional
Pseudomonas aeruginosa cytology
Pseudomonas aeruginosa physiology
Chromosomes, Bacterial genetics
DNA Transposable Elements genetics
Genetic Vectors
Green Fluorescent Proteins metabolism
Host-Pathogen Interactions
Pseudomonas aeruginosa genetics
Subjects
Details
- Language :
- English
- ISSN :
- 0008-4166
- Volume :
- 54
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Canadian journal of microbiology
- Publication Type :
- Academic Journal
- Accession number :
- 18388971
- Full Text :
- https://doi.org/10.1139/w07-118