Green fluorescent protein as a detection marker for Coxiella burnetii transformation.
The molecular biological study of the obligate intracellular bacterium Coxiella burnetii is hampered because of the lack of an efficient DNA transformation system. We used expression of the green fluorescent protein (GFP) in addition to ampicillin resistance as a selection marker for detection of transformed C. burnetii cells. Fluorescent microscopy studies revealed that transformed C. burnetii cells can be detected easily inside the host cell line. A high level of GFP expression was reached with the strong Escherichia coli trc (trp/lac) promoter. The use of GFP not only provides a convenient marker for transformation of C. burnetii, but also allows detection of this obligate intracellular pathogen inside host eukaryotic cells. Possible applications for GFP in the study of host-pathogen interactions are discussed.
Lukácová
M
M
Institute of Molecular Biology, Slovak Academy of Sciences, Bratislava, Slovak Republic.
Valková
D
D
Quevedo Diaz
M
M
Perecko
D
D
Barák
I
I
D000668:Ampicillin Resistance / Q000235:genetics; D000818:Animals; D015139:Blotting, Southern; D016997:Coxiella burnetii / Q000187:drug effects / Q000235:genetics*; D018274:Electroporation / Q000379:methods; D005819:Genetic Markers; D049452:Green Fluorescent Proteins; D007739:L Cells; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D010957:Plasmids; D016133:Polymerase Chain Reaction; D014169:Transformation, Bacterial
D005819:Genetic Markers; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins