Biomimetic Sensor for Pathogenic Bacteria
Abstract
This study investigated the assembly of a biomimetic sensor containing an osmotic receptor protein. The first objective of this research included the assembly and immobilization of fluorescently tagged liposomes. The second objective involved the expression and purification of an osmotic sensitive protein (MscL) and the incorporation of this protein into the liposome membrane. Liposomes (2 micron diameter) containing fluorescein labeled phospholipids and biotinylated phosphotidyl ethanolamine in the membranes and internalized soluble sulforhodamine were assembled. Liposomes were characterized with respect to composition, size, and shelf-life using confocal microscopy. Avidin was covalently attached to a glass surface for the immobilization of the biotinylated liposomes. Immobilization of fluorescent liposomes was confirmed with confocal microscopy. The liposomes contain a green/yellow lipid bilayer and a red interior. The cloning of recombinant MScL into an Escherichia coli expression system yielded an MScL-thioredoxin fusion protein that was tagged with a blue fluorescent dye and incorporated into the membrane of the liposomes. The functionality of the proteosome was observed by the release of the water soluble sulforhodamine in the presence of high salt, 3 M, concentrations.
Document Details
- Document Type
- Technical Report
- Publication Date
- Mar 05, 2001
- Accession Number
- ADA388146
Entities
People
- Bart C. Weimer
- Daryll B. De Wald
- Marie K. Walsh
Organizations
- Utah State University