Detection of oxytetracycline production by Streptomyces rimosus in soil microcosms by combining whole-cell biosensors and flow cytometry.
Combining the high specificity of bacterial biosensors and the resolution power of fluorescence-activated cell sorting (FACS) provided qualitative detection of oxytetracycline production by Streptomyces rimosus in soil microcosms. A plasmid containing a transcriptional fusion between the tetR-regulated P(tet) promoter from Tn10 and a FACS-optimized gfp gene was constructed. When harbored by Escherichia coli, this plasmid produces large amounts of green fluorescent protein (GFP) in the presence of tetracycline. This tetracycline biosensor was used to detect the production of oxytetracycline by S. rimosus introduced into sterile soil. The tetracycline-induced GFP-producing biosensors were detected by FACS analysis, enabling the detection of oxytetracycline encounters by single biosensor cells. This approach can be used to study interactions between antibiotic producers and their target organisms in soil.
Hansen
L H
LH
Department of General Microbiology, University of Copenhagen, DK-1307 Copenhagen K, Denmark.
Ferrari
B
B
Sørensen
A H
AH
Veal
D
D
Sørensen
S J
SJ
D015374:Biosensing Techniques / Q000379:methods*; D003470:Culture Media; D017753:Ecosystem; D005434:Flow Cytometry / Q000379:methods*; D010118:Oxytetracycline / Q000096:biosynthesis*; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012988:Soil Microbiology; D013302:Streptomyces / Q000254:growth & development / Q000378:metabolism*; D013752:Tetracycline
D003470:Culture Media; D011993:Recombinant Fusion Proteins; D013752:Tetracycline; D010118:Oxytetracycline