Cloning of a region in the Brucella abortus chromosome necessary for o-side chain biosynthesis

Simple item page
dc.contributor.authorMcQuiston, John R.en
dc.contributor.departmentVeterinary Medical Sciencesen
dc.date.accessioned2014-03-14T21:43:29Zen
dc.date.adate2009-08-22en
dc.date.available2014-03-14T21:43:29Zen
dc.date.issued1992en
dc.date.rdate2009-08-22en
dc.date.sdate2009-08-22en
dc.description.abstractAs a first step in characterizing the genes involved in O-side chain synthesis in <i>Brucella abortus</i> strain 2308, a portion of the genomic DNA was cloned from a rough mutant created by Tn5 (KnR) mutagenesis. This mutant was rough based on the lack of reactivity by either whole cells or extracted LPS to an O-side chain monoclonal antibody (BRU-38). A 30 kb <i>Xba</i>I genomic fragment (including Tn5) from the rough strain was subcloned into a sequencing vector to create pJM6. When <i>B. abortus</i> 2308 was electroporated with pJM6, KnR clones were unable to react with BRU-38; a Southern analysis of these clones revealed Tn5 in the 30 kb <i>Xba</i>I genomic fragment. Various regions of the 30kb fragment were subcloned and tested for their ability to complement specific <i>rfa</i> and <i>rfb</i> mutants of <i>Escherichia coli</i> and <i>Salmonella typhimurium</i>. One particular DNA fragment complemented an <i>rfbD</i> mutation in <i>E. coli</i> as judged by agglutination with <i>E. coli</i> anti-O (0:85) serum. The same DNA fragment failed to cause <i>E. coli rfbD</i> to react with either BRU-38 or <i>B. abortus</i> anti-O polyclonal antisera. The <i>B. abortus</i> 30 kb <i>Xba</i>I fragment contains a gene which has been identified by comple-mentation as containing the equivalent of the <i>rfbD</i> gene encoding dTDP-rhamnose synthetase in <i>E. coli</i>. Since <i>Brucella</i> is not known to have rhamnose in its core this enzyme may have a different function in <i>Brucella</i> LPS synthesis.en
dc.description.degreeMaster of Scienceen
dc.format.extentvi, 56 leavesen
dc.format.mediumBTDen
dc.format.mimetypeapplication/pdfen
dc.identifier.otheretd-08222009-040429en
dc.identifier.sourceurlhttp://scholar.lib.vt.edu/theses/available/etd-08222009-040429/en
dc.identifier.urihttp://hdl.handle.net/10919/44396en
dc.language.isoenen
dc.publisherVirginia Techen
dc.relation.haspartLD5655.V855_1992.M438.pdfen
dc.relation.isformatofOCLC# 27689383en
dc.rightsIn Copyrighten
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/en
dc.subject.lccLD5655.V855 1992.M438en
dc.subject.lcshBiosynthesisen
dc.subject.lcshBrucella abortus -- Geneticsen
dc.subject.lcshClone cellsen
dc.titleCloning of a region in the Brucella abortus chromosome necessary for o-side chain biosynthesisen
dc.typeThesisen
dc.type.dcmitypeTexten
thesis.degree.disciplineVeterinary Medical Sciencesen
thesis.degree.grantorVirginia Polytechnic Institute and State Universityen
thesis.degree.levelmastersen
thesis.degree.nameMaster of Scienceen

Files

Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
LD5655.V855_1992.M438.pdf
Size:
8.26 MB
Format:
Adobe Portable Document Format
Description:
Download

Collections

Masters Theses