Construction of Self-Transmissible Green Fluorescent Protein-Based Biosensor Plasmids and Their Use for Identification of N-Acyl Homoserine-Producing Bacteria in Lake Sediments

• Published in: Applied and Environmental Microbiology 2010-09, Vol.76 (18), p.6119-6127
• Main Authors: Lumjiaktase, Putthapoom, Aguilar, Claudio, Battin, Tom, Riedel, Kathrin, Eberl, Leo
• Format: Article
• Language: English
• Subjects: 4-Butyrolactone - analogs & derivatives; 4-Butyrolactone - metabolism; Bacteria; Bacteria - genetics; Bacteria - isolation & purification; Bacteria - metabolism; Base Sequence; Biofilms; Biological and medical sciences; Biosensing Techniques - methods; Biosensors; Biotechnology; Environmental Microbiology; Fluorescence; Fresh Water; Fundamental and applied biological sciences. Psychology; Geologic Sediments - microbiology; Green Fluorescent Proteins - biosynthesis; Lakes; Methods. Procedures. Technologies; Microbiology; Molecular Sequence Data; Plasmids - genetics; Proteins; Proteobacteria; Quorum Sensing - physiology; RNA, Ribosomal, 16S - genetics; Sediments; Sequence Analysis, DNA; Switzerland; Various methods and equipments
• ISSN: 0099-2240; 1098-5336; 1098-6596
• DOI: 10.1128/AEM.00677-10

Many bacteria utilize quorum sensing (QS) systems to communicate with each other by means of the production, release, and response to signal molecules. N-Acyl homoserine lactone (AHL)-based QS systems are particularly widespread among the Proteobacteria, in which they regulate various functions. It has become evident that AHLs can also serve as signals for interspecies communication. However, knowledge on the impact of AHLs for the ecology of bacteria in their natural habitat is scarce, due mainly to the lack of tools that allow the study of QS in bacterial communities in situ. Here, we describe the construction of self-mobilizable green fluorescent protein (GFP)-based AHL sensors that utilize the conjugation and replication properties of the broad-host-range plasmid RP4. We show that these novel AHL sensor plasmids can be easily transferred to different bacterial species by biparental mating and that they give rise to green fluorescent cells in case the recipient is an AHL producer. We also demonstrate that these sensor plasmids are capable of self-spreading within mixed biofilms and are a suitable tool for the identification of AHL-producing bacteria in lake sediment.