The Characterisation of Putative Nuclear Pore-Anchoring Proteins in Arabidopsis thaliana

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dc.contributor.authorCollins, Patrick
dc.date.accessioned2014-02-24T20:53:48Z
dc.date.available2015-09-04T12:20:05Z
dc.date.issued2013en
dc.description.abstractThe nuclear pore complex (NPC) is perhaps the largest protein complex in the eukaryotic cell, and controls the movement of molecules across the nuclear envelope. The NPC is composed of up to 30 proteins termed nucleoporins (Nups), each grouped in different sub-complexes. The transmembrane ring sub-complex is composed of Nups responsible for anchoring the NPC to the nuclear envelope. Bioinformatic analysis has traced all major sub-complexes of the NPC back to the last eukaryotic common ancestor, meaning that the nuclear pore structure and function is conserved amongst all eukaryotes. In this study Arabidopsis T-DNA knockout lines for these genes were investigated to characterise gene function. Differences in plant growth and development were observed for the ndc1 knockout line compared to wild-type but gp210 plants showed no phenotypic differences. The double knockout line gp210 ndc1 was generated through crosses to observe plant response to the knockout of two anchoring-Nup genes. No synergistic affect from this double knockout was observed, suggesting that more, as yet unidentified Nups function the transmembrane ring in plants. The sensitivity to nuclear export inhibitor leptomycin B (LMB) was tested also for knockout lines, although growth sensitivity to the drug was not observed. Nucleocytoplasmic transport of knockout lines was measured in cells transformed by particle bombardment. To express fluorescent protein constructs actively transported through the NPC, localisation of protein determined the nucleocytoplasmic transport of the cell. The ndc1single knockout and the double knockout gp210 ndc1 exhibited decreased nuclear export. Further experiments in determining NDC1 localisation and identification of other Nups in the transmembrane ring sub-complex would bring a more comprehensive understanding to the plant NPC.en
dc.identifier.urihttp://hdl.handle.net/10092/8885
dc.identifier.urihttp://dx.doi.org/10.26021/9042
dc.language.isoen
dc.publisherUniversity of Canterbury. Biological Sciencesen
dc.relation.isreferencedbyNZCUen
dc.rightsCopyright Patrick Collinsen
dc.rights.urihttps://canterbury.libguides.com/rights/thesesen
dc.subjectArabidopsis thalianaen
dc.subjecttransformationen
dc.subjectinserten
dc.subjecttransient expressionen
dc.subjectepidermal cellsen
dc.subjectroot cellsen
dc.subjectgeneen
dc.subjectproteinen
dc.subjectplanten
dc.subjectboltingen
dc.subjectfloweringen
dc.subjectroot growthen
dc.subjectknockouten
dc.subjectndc1en
dc.subjectgp210en
dc.subjectAt1g73240en
dc.subjectAt5g40480en
dc.subjectSALKen
dc.subjectSAILen
dc.subjectColumbiaen
dc.subjectseeden
dc.subjectgerminationen
dc.subjectDNA sequencingen
dc.subjectagarose gel electrophoresis confocal microscopyen
dc.subjectlight microscopyen
dc.subjectstereo-fluorescence microscopyen
dc.subjectDNA extractionen
dc.subjectcellen
dc.subjectnucleusen
dc.subjectcytoplasmen
dc.subjectnucleoplasmen
dc.subjectnuclear pore complexen
dc.subjectnucleoporinen
dc.subjectnuclear envelopeen
dc.subjectputativeen
dc.subjectnuclear pore-anchoring proteinen
dc.subjectleptomycin Ben
dc.subjectgene gunen
dc.subjectparticle bombardmenten
dc.subjectagrobacteriumen
dc.subjectT-DNAen
dc.subjectfasciationen
dc.subjectrhodococcus fasciansen
dc.subjectcrossen
dc.subjectreverse geneticsen
dc.subjectnucleocytoplasmic transporten
dc.subjectcrossen
dc.subjectselfen
dc.subjectsynergisticen
dc.subjectsiliqueen
dc.subjecttransmembrane ringen
dc.subjectdihybriden
dc.subjectpumilo homology domain proteinen
dc.subjectGFPen
dc.subjectYFPen
dc.subjectRFPen
dc.subjectPCRen
dc.subjecthomozygoteen
dc.subjectheterozygoteen
dc.subjectinhibitionen
dc.titleThe Characterisation of Putative Nuclear Pore-Anchoring Proteins in Arabidopsis thalianaen
dc.typeTheses / Dissertations
thesis.degree.disciplinePlant Biotechnologyen
thesis.degree.grantorUniversity of Canterburyen
thesis.degree.levelMastersen
thesis.degree.nameMaster of Scienceen
uc.bibnumber1952880
uc.collegeFaculty of Scienceen
uc.embargo24en
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