Bacteroides Fragilis OmpA: Utility as a Live Vaccine Vector for Biodefense Agents
Abstract
We are studying the utility of using B.fragilis OmpA as a vehicle on which to put antigenic epitopes of organisms that can be used in bioterror, with the aim of eventually constructing a vaccine vehicle vector. OmpA is the major outer membrane protein of B. fragilis, a gram negative anaerobe that normally resides in the gut. There are four homologs for ompA in the genome. The purpose of this study was to construct a B. fragilis ompA deletant and to begin to characterize the function of OmpA, as well as the specific function(s) of the various loops exposed on the surface. Initial studies indicated difficulty in achieving stable expression, export to periplasm and integration of B. fragilis OmpA into E. coli membrane. Thus we optimized a method to insert the recombinant gene directly into B. fragilis. We inserted two restriction sites into a recombinant ompA gene so that we could later replace the internal portion of the gene with one modified to express specific epitopes on loop 2 and loop 3. We will proceed by expressing and detecting the FLAG-tagged OmpAs directly in B. fragilis WAL 186 ompA deletion strain, as well as characterizing the phenotypes of B. fragilis with modified OmpAs.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 01, 2008
- Accession Number
- ADA486636
Entities
People
- Hannah M. Wexler