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dc.contributor.authorModise, Theroen_US
dc.date.accessioned2016-09-07T08:00:25Z
dc.date.available2016-09-07T08:00:25Z
dc.date.issued2016-09-06en_US
dc.identifier.othervt_gsexam:8833en_US
dc.identifier.urihttp://hdl.handle.net/10919/72885
dc.description.abstractThe pathogen Francisella tularensis subsp. tularensis has been classified as a Center for Disease Control (CDC) select agent. However, little is still known of what makes the bacteria cause dis-ease, especially the highly virulent type A1 strains. The work in this dissertation focused on type A1 strains from the Inzana laboratory, including a wildtype virulent strain TI0902, an avirulent chemical mutant strain TIGB03 with a Single Nucleotide Polymorphism in the wbtK gene, and several complemented mutants, [wbtK+]TIGB03, with dramatic differences in virulence and growth rates. One of the complemented clones (Clone12 or avp-[wbtK+]TIGB03-C12) was aviru-lent, but protected mice against challenge of a lethal dose of TI0902 and was considered as a possible vaccine strain. Whole genome sequencing was performed to identify genetic differences between the virulent, avirulent and protective strains using both Roche/454 and Illumina next-generation sequencing technologies. Additionally, RNASeq analysis was performed to identify differentially expressed genes between the different strains. This comprehensive genomic analysis revealed the critical role of transposable elements in inducing genomic instability resulting in large du-plications and deletions in the genomes of the chemical mutant and the complemented clones that in turn affect gene dosage and expression of genes known to regulate virulence. For exam-ple, whole genome sequencing of the avirulent chemical mutant TIGB03 revealed a large 75.5 kb tandem duplication flanked by transposable elements, while the genome of a virulent Clone01 (vir-[wbtK+]TIGB03-C1) lost one copy of the 75.5 kb tandem duplicated region but gained a tandem duplication of another large 80kb region that contains a virulence associated transcription factor SspA. RNAseq data showed that the dosage effect of this extra region in Clone1 suppresses expression of MglA regulated genes. Since MglA regulates genes that are known to be crucial for virulence, including the well-studied Francisella Pathogenicity Island (FPI), these results suggest that gene dosage effects arising from large duplications can trigger unknown virulence mechanisms in F. tularensis subsp. tularensis. These results are important especially when designing live vaccine strains that have repeated insertion elements in their genomes.en_US
dc.format.mediumETDen_US
dc.publisherVirginia Techen_US
dc.rightsThis Item is protected by copyright and/or related rights. Some uses of this Item may be deemed fair and permitted by law even without permission from the rights holder(s), or the rights holder(s) may have licensed the work for use under certain conditions. For other uses you need to obtain permission from the rights holder(s).en_US
dc.subjectFrancisella tularensisen_US
dc.subjectgenome instabilityen_US
dc.subjectvaccineen_US
dc.subjecttransposaseen_US
dc.subjectinversionen_US
dc.subjectduplica-tionen_US
dc.subjectdeletionen_US
dc.subjectRNAseqen_US
dc.subjectDNAseqen_US
dc.subjectAssemblyen_US
dc.subjectGene Dosageen_US
dc.subjectPathogenen_US
dc.titleGenomic Instability and Gene Dosage Obscures Clues to Virulence Mechanisms of F. tularensis speciesen_US
dc.typeDissertationen_US
dc.contributor.departmentAnimal and Poultry Sciencesen_US
dc.description.degreePHDen_US
thesis.degree.namePHDen_US
thesis.degree.leveldoctoralen_US
thesis.degree.grantorVirginia Polytechnic Institute and State Universityen_US
thesis.degree.disciplineGenetics, Bioinformatics, and Computational Biologyen_US
dc.contributor.committeechairJensen, Roderick Ven_US
dc.contributor.committeememberInzana, Thomas Josephen_US
dc.contributor.committeememberHeath, Lenwood Sen_US
dc.contributor.committeememberGood, Deborah Jen_US


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