The in vivo localization and interactions between structural components of the Shewanella oneidensis MR-1 metal reducing system by high resolution transmission electron microscopy.
EMSL Project ID
18594
Abstract
To understand the mechanisms of anaerobic metal oxide reduction, important issues regarding the subcellular localization and the outer membrane architecture of the iron-reducing respiratory chain need to be resolved. To address these questions, we have generated polyclonal antibodies specifically recognizing different epitopes of GspG, MtrC, MtrA, OmcA and MtrB proteins. Sera generated towards surface exposed epitopes of these proteins will be used to determine the subcellular localization of these proteins using immunogold-labeling methods and high resolution transmission electron microscopy. Immuno-gold labeling will be preformed on ultra-thin frozen sections or whole mounts of steady state cell cultures. For localization of proteins associated with bacterial structures, cells will be fixed, embedded in gelatin, and flash frozen. The ultra thin cryosections will be obtained, mounted on a grid, and incubated using our established immuno-EM hybridization protocols. These tasks will be coordinated with other members of the BGC team including Dr. Terry Beveridge (electron microscopy) at the University of Guelph, Ontario, Canada.
Project Details
Project type
Grand Challenge
Start Date
2006-03-24
End Date
2008-10-01
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Marshall MJ, A Dohnalkova, DW Kennedy, AE Plymale, SH Thomas, FE Loffler, R Sanford, JM Zachara, JK Fredrickson, and AS Beliaev. 2009. "Electron donor-dependent radionuclide reduction and nanoparticle formation by Anaeromyxobacter dehalogenans strain 2CP-C." Environmental Microbiology 11(2):534-543.