Progressive myoclonus epilepsy type 1 (EPM1), also known as Unverricht-Lundborg disease, is one of the rare forms of epilepsy that shows a clear pattern of autosomal recessive inheritance. The gene defective in this disease was linked to the distal tip of chromosome 21, in band q22.3. In this study, we have collected 93 samples from 15 EPM1 families and 5 affected individuals as the basis for identifying the EPM1 gene. We have also constructed a 770 kb cosmid and bacterial artificial chromosome contig covering the candidate EPM1 region, and have isolated expressed sequences from this contig. For three of the genes that we isolated (GT335, GT334, and PWP2), we have identified and sequenced a full-length cDNA, identified the putative protein, assessed the expression pattern of the gene by Northern blot, determined the exon/intron structure of the gene, characterized basepair polymorphisms within each gene, and finally excluded each of these genes as the one defective in EPM1 patients. Using these new polymorphisms, and others that were available and that we had identified, we were able to construct detailed haplotypes on each of the affected EPM1 chromosomes, to help pinpoint, the location of the EPM1 gene, and help estimate the number of different mutations we might have in our collection. / While these studies were underway, another group identified the cystatin B (STFB) gene as that defective in EPM1. This allowed us to directly test this gene for mutations in our collection of EPM1 patients. We could identify four different mutations in the STFB gene, the most common of which consisted of a variable length insertion in the 5 ' flanking region of the gene, and which was previously undescribed. This mutation, which is found in 78% of unrelated EPM1 chromosomes we studied, showed some level of meiotic instability, and mapped to a polymorphic 12-bp GC-rich repeat. Using a combination of PCR and Southern blotting assays, we could accurately diagnose nearly 100% of all EPM1 patients. This represents a significant step forward in our ability to diagnose this disease at the molecular level, and should allow a more precise definition of the progressive myoclonus epilepsies, as a whole.

Obligate shade plants possess adaptations that enable them to photosynthesise in the low light environment of the forest floor. Adaptations that facilitate light scavenging may compromise capacity for high rates of photosynthesis. This study compares the responses of obligate shade and facultative shade plant species upon exposure to elevated light. The obligate shade plants were two commercially grown medicinal herb species of ginseng, Panax ginseng C.A. Meyer and Panax quinquefolius L.; and goldenseal - Hydrastis canadensis L. Comparison was made to Arabidopsis thaliana and Pisum sativum L. as facultative shade species. Panax ginseng (Korean ginseng) and Panax quinquefolius (American ginseng) are obligate shade plants found in broadleaf forests of Eastern Asia and North America, respectively. Studies on these plants have shown optimal growth at light intensities between 200-300 [mu]mol photons. m⁻�. s⁻�, or 10-15% of full sunlight, and at intensities greater than 500 [mu]mol photons. m⁻�. s⁻� characteristic photoinbibitory symptoms develop. An atypical response to methyl viologen in photosynthetic electron transport assays was observed in ginseng in both isolated thylakoid membranes and whole leaves. No correlation was found between detectable superoxide dismutase activity and altered methyl viologen reactions. In a mutagenesis study using the model cyanobacterium Synechocystis sp. PCC 6803, a unique amino acid residue in the terminal electron acceptor PsaC, found only in ginseng, was changed and found to have no effect on methyl viologen reactions. Electron transfer to methyl viologen was examined in both isolated thylakoid membranes and whole leaves using chlorophyll a fluorescence and the apparent ability for methyl viologen to act as an electron acceptor was observed to differ between ginseng species. Obligate shade species were observed to possess alternate pools of photosystem II centres that potentially provide a mechanism to maximise photosynthetic gain under low light and during short periods of increased illumination. In experiments designed to identify physiological processes that contribute to increased susceptibility to photoinhibition in obligate shade plants, responses were observed and characterised following a moderate increase in illumination (140 to 400 [mu]mol photons. m⁻� . s⁻�) using chlorophyll a fluorescence induction curve analysis. The obligate shade species exhibited varied responses to elevated light and showed increased susceptibility, to photoinhibition. Photoprotective non-photochemical dissipative capacity was quantified and found to be comparable between all species studied.

Includes bibliographical references (leaves 196-229). / 229 leaves : ill. (some col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / A mutant lacking Vibrio cholerae lipoprotein A activity has been constructed in V. cholerae O139 which contains two copies of vlpA, by allelic exchange after inactivation via the insertion of kanamycin and tetracycline resistance gene cartridges. The vlpA-negative mutant has been assessed for virulence in the infant mouse cholera model. / Thesis (Ph.D.)--Adelaide University, Dept. of Molecular Biosciences, 2001

"February 1977." / Includes bibliographical references (leaves [105]-[120]) / xiv, 104, [16] leaves : ill. ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / The malignant transformation, and the relevance of developmental antigens to the cancer process, is broadly reviewed. The two developmental antigens - foetal and placental - are then examined in experimental mouse and rat tumour systems. / Thesis (Ph.D.)--University of Adelaide, Dept. of Animal Physiology, 1978

Includes bibliographical references (leaves 196-229). / 229 leaves : ill. (some col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / A mutant lacking Vibrio cholerae lipoprotein A activity has been constructed in V. cholerae O139 which contains two copies of vlpA, by allelic exchange after inactivation via the insertion of kanamycin and tetracycline resistance gene cartridges. The vlpA-negative mutant has been assessed for virulence in the infant mouse cholera model. / Thesis (Ph.D.)--Adelaide University, Dept. of Molecular Biosciences, 2001

No description available.

Li, Kwun Yee. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 83-88). / Abstract also in Chinese. / Thesis/Assessment Committee --- p.ii / Statement --- p.iii / Abstract --- p.iv / 摘要 --- p.vi / Acknowledgements --- p.vii / Table of Contents --- p.ix / List of Tables --- p.xiii / List of Figures --- p.xiv / List of Abbreviations --- p.xvii / Chapter Chapter 1 --- General Introduction --- p.1 / Chapter 1.1 --- The Plant Secretory and Endocytic Pathways --- p.2 / Chapter 1.2 --- PVC Proteomics Analysis Led to the Identification of AtEMP --- p.5 / Chapter 1.3 --- EMP70 Family Proteins --- p.5 / Chapter 1.3.1 --- General structure of EMP70 proteins --- p.5 / Chapter 1.3.2 --- EMP70 in other organisms --- p.8 / Chapter 1.3.3 --- EMP70 proteins in Arabidopsis --- p.9 / Chapter 1.4 --- Accession Numbers --- p.10 / Chapter 1.5 --- Research Objectives --- p.14 / Chapter Chapter 2 --- Generation and Characterization of Transgenic Tobacco BY-2 Cell Lines Expressing Selective AtEMP-GFP Fusions --- p.15 / Chapter 2.1 --- Introduction --- p.16 / Chapter 2.2 --- Materials and Methods --- p.17 / Chapter 2.2.1 --- RNA extraction and cDNA generation --- p.17 / Chapter 2.2.2 --- Construct making --- p.18 / Chapter 2.2.3 --- Bacterial strains --- p.21 / Chapter 2.2.4 --- Transformation of BY-2 cells --- p.21 / Chapter 2.2.5 --- Confocal fluorescence screening of tobacco BY-2 cells --- p.23 / Chapter 2.2.6 --- Drug treatments --- p.23 / Chapter 2.3 --- Results --- p.25 / Chapter 2.3.1 --- Western blot analysis of tobacco BY-2 cell lines expressing AtEMP-GFP fusions --- p.25 / Chapter 2.3.2 --- Subcellular localization of AtEMP-GFP fusions to the PVC in transgenic BY-2 cells --- p.27 / Chapter 2.4 --- Summary --- p.30 / Chapter Chapter 3 --- Generation and Characterization of Antibodies Against Various AtEMPs --- p.31 / Chapter 3.1 --- Introduction --- p.32 / Chapter 3.2 --- Materials and Methods --- p.33 / Chapter 3.2.1 --- Generation of antibodies --- p.33 / Chapter 3.2.2 --- Screening of antibodies --- p.36 / Chapter 3.2.2.1 --- SDS-PAGE and western blot analysis --- p.36 / Chapter 3.2.2.2 --- Confocal immunofluorescence studies --- p.38 / Chapter 3.3 --- Results --- p.39 / Chapter 3.3.1 --- AtEMP antibodies recognized EMP70 proteins in plant cells --- p.39 / Chapter 3.3.2 --- Organelles marked by anti-AtEMPs are closely associated with the Golgi apparatus --- p.40 / Chapter 3.4 --- Summary --- p.49 / Chapter Chapter 4 --- Subcellular Localization of GFP-tagged AtEMP Fusions via Transient Expression --- p.50 / Chapter 4.1 --- Introduction --- p.51 / Chapter 4.2 --- Materials and Methods --- p.52 / Chapter 4.2.1 --- Making of transient expression constructs --- p.52 / Chapter 4.2.2 --- Transient expression --- p.57 / Chapter 4.3 --- Results --- p.59 / Chapter 4.3.1 --- PVC localization of AtEMP-GFP fusions --- p.59 / Chapter 4.3.2 --- Golgi localization of GFP-AtEMP and GFP-AtEMP-S fusions --- p.62 / Chapter 4.4 --- Summary --- p.66 / Chapter Chapter 5 --- Immunogold Electron Microscope Localization of AtEMPs --- p.67 / Chapter 5.1 --- Introduction --- p.68 / Chapter 5.2 --- Materials and Methods --- p.68 / Chapter 5.2.1 --- High-pressure freezing / freeze substitution --- p.68 / Chapter 5.2.2 --- Ultra-thin sectioning --- p.69 / Chapter 5.2.3 --- Immunogold labeling --- p.69 / Chapter 5.2.4 --- Post-staining and transmission election microscopy --- p.69 / Chapter 5.3 --- Results and Summary --- p.70 / Chapter Chapter 6 --- Discussion and Future Perspectives --- p.74 / Chapter 6.1 --- Hypothesis --- p.75 / Chapter 6.2 --- Subcellular localization of AtEMPs --- p.76 / Chapter 6.2.1 --- GFP-tagged AtEMP fusions --- p.76 / Chapter 6.2.2 --- Endogenous EMP70 proteins in BY-2 cells --- p.77 / Chapter 6.3 --- Targeting motifs in AtEMPs --- p.79 / Chapter 6.4 --- Conclusions --- p.81 / Chapter 6.5 --- Future perspectives --- p.82 / Chapter 6.5.1 --- Targeting motifs --- p.82 / Chapter 6.5.2 --- Functional studies --- p.82 / References --- p.83

The primary aim of this project was to examine factors which are likely to provide a mechanism by which the polymorphism for capitulum type and associated outcrossing rate in Senecio vulgaris may be maintained. The majority of studies conducted examined the demographic genetics (i.e. the changes in the number of individuals of the two morphs at different life history stages) in field experiments initiated either in Spring or Autumn, The first series examined the demography of each morph raised from seedlings to senescence in pure stand and mixture. The second series examined the demography of each morph from seed to senescence and thereby investigated the effect on fitness of any difference between morphs in germination behaviour. The importance of inbreeding depression on the maintenance of the polymorphism was examined by comparing the relative fitness of self and open pollinated offspring of each morph under glasshouse conditions. Germination behaviour of seeds of each morph was also investigated in a series of field trials conducted over an extended period. These field studies were complemented by a series of synchronous laboratory studies to examine the effect of temperature on morph germination behaviour. It was found that inbreeding depression is not an important factor in the maintenance of the polymorphism. No short term advantage of the radiate over the non-radiate morph was evident in the first series of demography experiments. The germination studies showed that differences between morphs in germination behaviour may occur frequently in autumn sown seed. The second series of demography experiments showed that under certain conditions this difference in germination can lead to the radiate morph attaining a greater relative fitness than the non-radiate morph. Temperature was found to be a major factor controlling the initial dormancy of seeds after sowing. It is concluded that the difference between morphs in germination behaviour is the most likely factor that could maintain the polymorphism for capitulum type in Edinburgh populations of 8. vulgaris.

Ph.D. / Pimelea Banks & Sol. Ex Gaertn. nom. cons. is a large genus consisting of 110 species, of which 90 species are endemic to Australia, 19 to New Zealand and one to Lord Howe Island. The genus has a great diversity of life forms, breeding systems and habitat. Its closest related genus is Thecanthes Wikstr. Thecanthes comprises five species of annual herbs occurring in the Philippines, New Ireland and northern Australia. Australasian Thecanthes and Pimelea are the only genera within sub-tribe Pimeleinae (angiosperm family Thymelaeaceae) and are characterised by the reduction to two stamens. Here I present the most comprehensive molecular phylogenetic study for Pimelea and Thecanthes. Sequences data from nuclear ITS rDNA and plastid rbcL, rps16, matK and trnL-F intergeneric spacer were used to reconstruct a phylogeny for these genera. I have produced 457 new DNA sequences (five genes and 150 taxa) for the present analyses. The resulting phylogeny was used to assess the taxonomic status of Thecanthes and to evaluate the relationships with Pimeleinae since previous studies indicated a close relationship between Pimelea, Thecanthes and species of Gnidia L. from tropical Africa. The morphological delimitation of sections within Pimelea, the biogeography and the radiation of the genus have been revaluated. Pimelea was found to be monophyletic. It was concluded that Pimelea and Thecanthes are congeneric; consequently a paper has been submitted transferring all species of Thecanthes into Pimelea and making the new combination Pimelea filifolia (Rye) Motsi & Rye. Data analysis revealed very low sequence variation within the subtribe Pimeleinae. This suggested a rapid radiation of the genera, which was confirmed by my molecular dating analyses. Based on molecular clock techniques, I calculated the following ages for the origin of Pimelea: 4.1 mya for New Zealand Pimelea spp. and 13.38 mya for other Pimelea spp. The molecular data also indicated that Pimelea and South Africa Gnidia have a direct common ancestor. I also show that the New Zealand Pimelea are derived and dispersed from Australian

No description available.