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Applied and Environmental Microbiology, January 2001, p. 239-244, Vol. 67, No. 1
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.1.239-244.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Detection of Oxytetracycline Production by Streptomyces rimosus in Soil Microcosms by Combining Whole-Cell Biosensors and Flow Cytometry

Lars Hestbjerg Hansen,1 Belinda Ferrari,2 Anders Hay Sørensen,1 Duncan Veal,2 and Søren Johannes Sørensen1,*

Department of General Microbiology, University of Copenhagen, DK-1307 Copenhagen K, Denmark,1 and Department of Biological Sciences, Macquarie University, Sydney, New South Wales 2109, Sydney, Australia2

Received 18 May 2000/Accepted 1 November 2000

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 Ptet 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.

* Corresponding author. Mailing address: Department of General Microbiology, University of Copenhagen, Sølvgade 83 H, DK-1307 Copenhagen K, Denmark. Phone: 45 35 32 20 53. Fax: 45 35 32 20 40. E-mail: sjs{at}mermaid.molbio.ku.dk.

Applied and Environmental Microbiology, January 2001, p. 239-244, Vol. 67, No. 1
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.1.239-244.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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