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dc.contributor.authorSiefkes-Boer, Henderika Jantina
dc.date.accessioned2010-04-19T22:08:18Z
dc.date.available2010-04-19T22:08:18Z
dc.date.issued1991
dc.identifier.urihttps://hdl.handle.net/10182/1695
dc.description.abstractThe similarities and differences between plants and other organisms can be studied by introducing non-plant regulatory sequences into the plant genome. Experiments were carried out to study gene expression under the regulation of an animal virus enhancer in faba bean (Vicia faba L. cv. Puma). A binary vector, pGA643-GUS-SV, was constructed carrying the cauliflower mosaic virus 35S (CaMV 35S) plant promoter, the Simian virus 40 (SV40) animal enhancer and the β-D glucuronidase (GUS) reporter gene. This plasmid was transferred to hairy root-inducing Agrobacterium tumefaciens strain A4T. Vicia faba L. cv. Puma plants were inoculated with A. tumefaciens A4T carrying pGA643-GUS-SV. Control V.faba L. cv. Puma plants were inoculated with A4T carrying the enhancer-less binary vector pGA643-GUS. The hairy roots were assayed for GUS expression and DNA content. Southern blots were carried out on these hairy roots to confirm GUS gene transfer and integration into V.faba L. cv. Puma, and to assess GUS copy numbers. The GUS activities showed high variability in both treatment pGA643-GUS and treatment pGA643-GUS-SV. This variability was reduced when the GUS activity values were adjusted by GUS copy numbers. The considerable amount of variability that remained after the adjustment was probably due to (i) differences in hairy root physiology and (ii) insert positioning effects in the plant genome. The mean GUS activity was slightly higher in treatment pGA643-GUS-SV than that in treatment pGA643-GUS. This difference, however, was not statistically significant at the 5% level, probably partly due to the variability observed in GUS activity values. The inability of the SV40 enhancer to significantly stimulate GUS gene expression in V.faba L. cv. Puma might have also been due to (i) the lack of an appropriate transcription factor or factors to bind to the enhancer, (ii) the nature of the intervening DNA between the enhancer and the promoter and (iii) possibly, to an unknown inhibiting factor present in V. jaba L. cv. Puma.en
dc.language.isoenen
dc.publisherLincoln Universityen
dc.rights.urihttps://researcharchive.lincoln.ac.nz/page/rights
dc.subjectregulatory sequencesen
dc.subjectgene expressionen
dc.subjectfaba beansen
dc.subjectVicia faba L. cv. Pumaen
dc.subjectbinary vectoren
dc.subjectCaMV 35S plant promoteren
dc.subjectSV 40 animal enhanceren
dc.subjectGUS reporter geneen
dc.subjectplasmiden
dc.subjectAgrobacterium tumejaciens A4Ten
dc.subjecthairy rootsen
dc.subjectgene transferen
dc.subjectgene integrationen
dc.titleGene expression in Vicia faba L. under regulation of the SV40 enhanceren
dc.typeThesisen
thesis.degree.grantorLincoln Universityen
thesis.degree.levelDoctoralen
thesis.degree.nameDoctor of Philosophyen
dc.subject.marsdenFields of Research::300000 Agricultural, Veterinary and Environmental Sciences::300200 Crop and Pasture Production::300203 Plant improvement (selection, breeding and genetic engineering)en
dc.subject.marsdenFields of Research::270000 Biological Sciences::270800 Biotechnologyen
dc.subject.marsdenFields of Research::270000 Biological Sciences::270800 Biotechnology::270803 Transgenesisen
lu.thesis.supervisorNoonan, Michael
lu.thesis.supervisorBullock, David
lu.contributor.unitDepartment of Wine, Food and Molecular Biosciencesen
dc.rights.accessRightsDigital thesis can be viewed by current staff and students of Lincoln University only. Print copy available for reading in Lincoln University Library. May be available through inter-library loan.en


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