{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T05:32:27Z","timestamp":1791178347888,"version":"4.1.0"},"reference-count":43,"publisher":"American Society for Microbiology","issue":"4","license":[{"start":{"date-parts":[[2002,4,1]],"date-time":"2002-04-01T00:00:00Z","timestamp":1017619200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2002,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            A green fluorescent protein-based\n            <jats:italic>Pseudomonas fluorescens<\/jats:italic>\n            strain A506 biosensor was constructed and characterized for its potential to measure benzene, toluene, ethylbenzene, and related compounds in aqueous solutions. The biosensor is based on a plasmid carrying the toluene-benzene utilization (\n            <jats:italic>tbu<\/jats:italic>\n            ) pathway transcriptional activator TbuT from\n            <jats:italic>Ralstonia pickettii<\/jats:italic>\n            PKO1 and a transcriptional fusion of its promoter\n            <jats:italic>PtbuA1<\/jats:italic>\n            with a promoterless\n            <jats:italic>gfp<\/jats:italic>\n            gene on a broad-host-range promoter probe vector. TbuT was not limiting, since it was constitutively expressed by being fused to the neomycin phosphotransferase (\n            <jats:italic>nptII<\/jats:italic>\n            ) promoter. The biosensor cells were readily induced, and fluorescence emission after induction periods of 3 h correlated well with toluene, benzene, ethylbenzene, and trichloroethylene concentrations. Our experiments using flow cytometry show that intermediate levels of\n            <jats:italic>gfp<\/jats:italic>\n            expression in response to toluene reflect uniform induction of cells. As the toluene concentration increases, the level of\n            <jats:italic>gfp<\/jats:italic>\n            expression per cell increases until saturation kinetics of the TbuT-PtbuA1 system are observed. Each inducer had a unique minimum concentration that was necessary for induction, with\n            <jats:italic>K<\/jats:italic>\n            <jats:sub>app<\/jats:sub>\n            values that ranged from 3.3 \u00b1 1.8 \u03bcM for toluene to 35.6 \u00b1 16.6 \u03bcM for trichloroethylene (means \u00b1 standard errors of the means), and maximal fluorescence response. The fluorescence response was specific for alkyl-substituted benzene derivatives and branched alkenes (di- and trichloroethylene, 2-methyl-2-butene). The biosensor responded in an additive fashion to the presence of multiple inducers and was unaffected by the presence of compounds that were not inducers, such as those present in gasoline. Flow cytometry revealed that, in response to toxic concentrations of gasoline, there was a small uninduced population and another larger fully induced population whose levels of fluorescence corresponded to the amount of effectors present in the sample. These results demonstrate the potential for green fluorescent protein-based bacterial biosensors to measure environmental contaminants.\n          <\/jats:p>","DOI":"10.1128\/aem.68.4.1962-1971.2002","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T09:59:40Z","timestamp":1027763980000},"page":"1962-1971","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":117,"title":["Development and Characterization of a Green Fluorescent Protein-Based Bacterial Biosensor for Bioavailable Toluene and Related Compounds"],"prefix":"10.1128","volume":"68","author":[{"given":"Lawrence","family":"Stiner","sequence":"first","affiliation":[{"name":"Microbiology, Iowa State University, Ames, Iowa 50011-1010"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Larry J.","family":"Halverson","sequence":"additional","affiliation":[{"name":"Departments of Agronomy"},{"name":"Microbiology, Iowa State University, Ames, Iowa 50011-1010"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.64.7.2730-2735.1998"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/0167-7799(93)90155-3"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.061014498"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.172.9.4749-4757.1990"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.178.21.6327-6337.1996"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.65.2.409-414.1999"},{"key":"e_1_3_2_8_2","unstructured":"Chalfie M. and S. 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