title: Development and Application of a Synthetically-Derived Lead Biosensor Construct for Use in Gram-Negative Bacteria.
abstract: The use of lead in manufacturing has decreased significantly over the last few decades. However, previous widespread use of lead-containing products and their incorrect disposal has resulted in environmental contamination. Accumulation of harmful quantities of lead pose a threat to all living organisms, through inhalation, ingestion, or direct contact, resulting in lead poisoning. This study utilized synthetic biology principles to develop plasmid-based whole-cell bacterial biosensors for detection of lead. The genetic element of the lead biosensor construct consists ofpbrR, which encodes the regulatory protein, together with its divergent promoter region and a promoterlessgfp. GFP expression is controlled by PbrR in response to the presence of lead. The lead biosensor genetic element was cloned onto a low-copy number broad host range plasmid, which can stably exist in a range of laboratory and environmental isolates, includingPseudomonas,Shewanella, andEnterobacter. The biosensors constructed were found to be sensitive, rapid, and specific and could, as such, serve as monitoring tools for lead-contaminated water.
authors.affiliation: Applied and Environmental Microbiology Laboratory, Department of Physiology, Anatomy and Microbiology, La Trobe University, Melbourne, Victoria 3086, Australia. ltbereza-malcolm@students.latrobe.edu.au.
authors.forename: Lara
authors.initials: L
authors.lastname: Bereza-Malcolm
authors.affiliation: Applied and Environmental Microbiology Laboratory, Department of Physiology, Anatomy and Microbiology, La Trobe University, Melbourne, Victoria 3086, Australia. S.Aracic@latrobe.edu.au.
authors.forename: Sanja
authors.initials: S
authors.lastname: Aracic
authors.affiliation: Applied and Environmental Microbiology Laboratory, Department of Physiology, Anatomy and Microbiology, La Trobe University, Melbourne, Victoria 3086, Australia. A.Franks@latrobe.edu.au.
authors.forename: Ashley E
authors.initials: AE
authors.lastname: Franks
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D004926:Escherichia coli / Q000302:isolation & purification; D006090:Gram-Negative Bacteria / Q000302:isolation & purification*; D057230:Limit of Detection; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000378:metabolism; D013997:Time Factors; D014874:Water Pollutants, Chemical / Q000032:analysis
chemical_list: D014874:Water Pollutants, Chemical
keywords: environmental monitoring; lead; synthetic biology; whole cell biosensors