Characterization of the structure and function of a Bacteroides thetaiotaomicron 16S rRNA promoter

Simple item page
dc.contributor.authorThorson, Mary Leahen
dc.contributor.committeechairStevens, Ann M.en
dc.contributor.committeememberChen, Jiann-Shinen
dc.contributor.committeememberMelville, Stephen B.en
dc.contributor.departmentBiologyen
dc.date.accessioned2014-03-14T21:37:20Zen
dc.date.adate2003-06-13en
dc.date.available2014-03-14T21:37:20Zen
dc.date.issued2003-06-06en
dc.date.rdate2004-06-13en
dc.date.sdate2003-06-09en
dc.description.abstractThe bacteroides group is a subdivision in the <I>Cytophaga-Flavobacterium-Bacteroides</I> phylum. This group is as phylogenetically distinct from other Gram-negative enterics, including <I>Escherichia coli</I>, as they are from Gram-positive organisms. Furthermore, there is no cross expression between genes of <I>E. coli</I> and <I>Bacteroides</I> species. It is thought that this difference in gene expression lies in part at the level of transcription initiation and is due to the sequences within the promoter region itself. A putative consensus sequence for <I>Bacteroides</I> promoters has been published by C. Jeff Smith&#146;s research group based on alignments of the sequences upstream of certain regulated genes. However, this consensus has not been found within all putative <I>Bacteroides</I> promoters. In this study, the promoter structure and function of a strong housekeeping <I>B. thetaiotaomicron</I> 16S rRNA promoter was examined and compared to an <I>E. coli</I> 16S rRNA promoter. Our hypothesis is that there are significant differences between the promoters of these two organisms. Analysis of <I>B. thetaiotaomicron</I> sequence upstream of the 16S rRNA gene has revealed the same overall structure known for <I>E. coli</I> 16S rRNA promoters in that there are two putative promoters separated by approximately 150 bp. However, the <I>B. thetaiotaomicron</I> 16S rRNA promoter contains the proposed <I>Bacteroides</I> &#151;7 and &#151;33 consensus sequences instead of the well known <I>E. coli</I> &#151;10 and &#151;35 consensus sequences. The biological activity of the<I> B. thetaiotaomicron</I> 16S rRNA full-length promoter was confirmed using a <I>Bacteroides lux</I> reporter system. A newly designed <I>Bacteroides lux</I> reporter was used to analyze specific regions of the <I>B. thetaiotaomicron</I> 16S rRNA promoter. In addition, by pairing the <I>B. thetaiotaomicron</I> 16S rRNA promoter with an <I>E. coli</I> ribosomal binding site, and vice-versa, the improved <I>lux</I> reporter was used to further confirm that the difference in gene expression between the two species lies at the level of transcription in <I>E. coli</I>. In <I>Bacteroides</I>, however, transcription and translation may work together to create a barrier to efficient gene expression of foreign genes. </P>en
dc.description.degreeMaster of Scienceen
dc.identifier.otheretd-06092003-130248en
dc.identifier.sourceurlhttp://scholar.lib.vt.edu/theses/available/etd-06092003-130248/en
dc.identifier.urihttp://hdl.handle.net/10919/42962en
dc.publisherVirginia Techen
dc.relation.haspartThesis2.PDFen
dc.rightsIn Copyrighten
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/en
dc.subjectrRNA operonen
dc.subjectpromoteren
dc.subjectgene reporteren
dc.subjectgene expressionen
dc.subjectBacteroidesen
dc.titleCharacterization of the structure and function of a <I>Bacteroides thetaiotaomicron</I> 16S rRNA promoteren
dc.typeThesisen
thesis.degree.disciplineBiologyen
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:
Thesis2.PDF
Size:
537.05 KB
Format:
Adobe Portable Document Format
Download

Collections

Masters Theses