Detection of Oxytetracycline Production byStreptomyces rimosusin Soil Microcosms by Combining Whole-Cell Biosensors and Flow Cytometry

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Abstract
Combining the high specificity of bacterial biosensors and the resolution power of fluorescence-activated cell sorting (FACS) provided qualitative detection of oxytetracycline production byStreptomyces rimosusin soil microcosms. A plasmid containing a transcriptional fusion between thetetR-regulated Ptetpromoter from Tn10and a FACS-optimizedgfpgene was constructed. When harbored byEscherichia 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 byS. rimosusintroduced 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.