Genetic engineering of Agrobacterium tumefaciens to target chloroplasts and identification of a novel nuclear localization signal of VirD2 protein

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dc.contributor.authorMatsuoka, Aki
dc.contributor.authorUniversity of Lethbridge. Faculty of Arts and Science
dc.contributor.supervisorZiemienowicz, Alicja
dc.contributor.supervisorKovalchuk, Igor
dc.date.accessioned2016-09-13T18:54:44Z
dc.date.available2016-09-13T18:54:44Z
dc.date.issued2015
dc.degree.levelPh.Den_US
dc.description.abstractAgrobacterium tumefaciens has been adopted to become a greatest tool for plant genetic engineering. Since Agrobacteria predominantly integrate single copy of the transgene into the nuclear genome, the transgene expression is relatively low. In contrast, transgene expression from chloroplasts allows high expression. Therefore, my first objective was to develop a new method of chloroplast transformation by modifying Agrobacterium genetics. I attempted to transform tobacco chloroplasts by Agrobacterium carrying the VirD2 protein with inactivated nuclear localization signals (NLSs) fused to a chloroplast transit peptide. However, the transformation of tobacco chloroplasts was unsuccessful. It suggested that the transit peptide was overridden by a novel cryptic NLS of VirD2. My second objective was to determine a novel NLS in VirD2. I discovered the novel NLS in VirD2 by subcellular localization analyses of fusion proteins (fragment of VirD2-green fluorescent protein) in plant cells. My discovery contributed to characterization of VirD2.en_US
dc.description.sponsorshipAlberta Innovates Technology Futuresen_US
dc.embargoYesen_US
dc.embargo.end2016-02-29
dc.embargo.start2015-09
dc.identifier.urihttps://hdl.handle.net/10133/4603
dc.language.isoen_CAen_US
dc.proquest.subject0307en_US
dc.proquestyesYesen_US
dc.publisherLethbridge, Alta : University of Lethbridge, Dept. of Biological Sciencesen_US
dc.publisher.departmentDepartment of Biological Sciencesen_US
dc.publisher.facultyArts and Scienceen_US
dc.relation.ispartofseriesThesis (University of Lethbridge. Faculty of Arts and Science)en_US
dc.subjectAgrobacterium-mediated transformationen_US
dc.subjectChloroplast transformationen_US
dc.subjectVirD2en_US
dc.subjectNuclear localization signalsen_US
dc.subjectTransit peptidesen_US
dc.subjectProtein diffusion through the nuclear poreen_US
dc.subjectSubcellular localizationen_US
dc.subjectProtein subcellular localization in leavesen_US
dc.titleGenetic engineering of Agrobacterium tumefaciens to target chloroplasts and identification of a novel nuclear localization signal of VirD2 proteinen_US
dc.typeThesisen_US
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