Show simple item record

dc.contributor.authorVemulapalli, Tracy H.en_US
dc.date.accessioned2014-03-14T20:31:40Z
dc.date.available2014-03-14T20:31:40Z
dc.date.issued2000-01-28en_US
dc.identifier.otheretd-02102000-20520015en_US
dc.identifier.urihttp://hdl.handle.net/10919/31194
dc.description.abstractBrucella abortus is a facultative, intracellular zoonotic pathogen, which can cause undulant fever in humans and abortion in cattle. Despite all of the progress in brucellosis research, there are still many unanswered questions regarding the molecular mechanisms involved in the pathogenesis of Brucella infections. To better understand the Brucella antigens involved in virulence and/or immunity, genetic and immunologic characterization of a 16 kDa protein of B. abortus was performed. Using PCR methods, the gene encoding the 16 kDa protein was cloned and sequenced. PCR and Southern blot analysis revealed that the gene is conserved among the 6 nomen species of Brucella. Overexpression of this protein in B. abortus vaccine strain RB51 was achieved using Brucella groE and sodC promoters as well as its own promoter. Protection and clearance studies were performed in mice to determine the role of this protein in Brucella immunity and pathogenesis. Inoculation with either strain RB51 overexpressing the 16 kDa protein or a DNA vaccine encoding the 16 kDa protein gene failed to provide significant protection. No difference was noted between the splenic clearance of B. abortus strain 2308 and its recombinant overexpressing the 16 kDa protein. A mutant of strain 2308 (2308D16) was created by disrupting the 16 kDa proteinâ s gene with a chloramphenicol resistance cassette. Western blot analysis indicated that the O antigen profile of strain 2308D16 differed from that of strain 2308. Mice cleared strain 2308D16 faster than strain 2308 indicating the potential attenuation of the disruption mutant. Purified 16 kDa protein was obtained by overexpressing it in E. coli via the pRSET expression system. Western blotting results initially identified this protein as an immunoglobulin-binding protein. Hemagglutination assay revealed that the 16 kDa protein exhibits lectin-like properties. Preliminary studies using hemagglutination inhibition identified mannose as a possible sugar to which the 16 kDa protein can interact. The lectin-like properties exhibited by the 16 kDa protein appears to influence smooth lipopolysaccharide production, and thereby may be involved in virulence.en_US
dc.publisherVirginia Techen_US
dc.relation.haspartThesis.pdfen_US
dc.rightsI hereby grant to Virginia Tech or its agents the right to archive and to make available my thesis or dissertation in whole or in part in the University Libraries in all forms of media, now or hereafter known. I retain all proprietary rights, such as patent rights. I also retain the right to use in future works (such as articles or books) all or part of this thesis or dissertation.en_US
dc.subjectBrucella abortusen_US
dc.subjectimmunoglobulin-binding proteinen_US
dc.subjectlectin-like proteinen_US
dc.titleGenetic and Immunological Analyses of a Brucella abortus Protein Exhibiting Lectin-like Propertiesen_US
dc.typeThesisen_US
dc.contributor.departmentBiomedical Sciences and Pathobiologyen_US
dc.description.degreeMaster of Scienceen_US
thesis.degree.nameMaster of Scienceen_US
thesis.degree.levelmastersen_US
thesis.degree.grantorVirginia Polytechnic Institute and State Universityen_US
thesis.degree.disciplineBiomedical Sciences and Pathobiologyen_US
dc.contributor.committeechairSriranganathan, Nammalwaren_US
dc.contributor.committeememberToth, Thomas E.en_US
dc.contributor.committeememberBoyle, Stephen M.en_US
dc.contributor.committeememberSchurig, Gerhardt G.en_US
dc.identifier.sourceurlhttp://scholar.lib.vt.edu/theses/available/etd-02102000-20520015/en_US
dc.date.sdate2000-02-10en_US
dc.date.rdate2001-02-16
dc.date.adate2000-02-16en_US


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record