Thesis (M. Sc.)--York University, 2002. Graduate Programme in Biology. / Title on thesis acceptance page : Molecular cloning of an [alpha]-tubulin from the Cestode, Hymenolepis, diminuta. Includes bibliographical references (leaves 110-125). Also available on the Internet. MODE OF ACCESS via web browser by entering the following URL: http://wwwlib.umi.com/cr/yorku/fullcit?pMQ71611.

published_or_final_version / Microbiology / Doctoral / Doctor of Philosophy

published_or_final_version / Zoology / Master / Master of Philosophy

published_or_final_version / Zoology / Master / Master of Philosophy

No description available.

No description available.

In order to clone the DNA repair gene of Aspergillus nidulans, uvsF$ sp-$ pyrG$ sp-$ strains were transformed with a genomic library in a plasmid vector that carried the pyr-4 gene of Neurospora which complements pyrG mutants of Aspergillus. Out of the several transformants obtained, four were like wild type. For rescuing plasmids, transformant DNA was digested with Bg/II and self ligated, and used for transformation of E. coli. Two types of plasmids were obtained; these two had a region in common ($<$1.0 kb) that was not a simple overlap and gave evidence for rearrangements. Surprisingly, only the plasmids with the larger insert of Aspergillus DNA were able to complement uvsF$ sp-$ in the secondary transformation. Northerns of polyA$ sp+$-enriched mRNA, probed with this plasmid, showed three bands. However, its subclone which spans the shared region hybridized to only one of them (1.0 kb). Two cDNA and five genomic clones were identified. The two cDNA clones though not identical, cross-hybridized. Three out of five genomic clones were identical. The cDNA hybridized to a short segment (2.2 kb) of one of the three types of genomic clones, locating the putative uvsF gene sequence.

Manual handling of biological cells is routine in most laboratories. This is gradually changing with the development of robotic cell handling systems, and micro-scale lab-on-chip devices. Attempts were made to develop devices that automate or assist cell handling in the context of a bovine nuclear transfer (NT) system. The system, a zona-free bovine NT cloning system, formed a baseline reference for tool design and performance evaluation. Bovine NT can, as other cell handling procedures, be improved by rapid and precise cell positioning. Improvements in cell handling can increase the quantity of cells processed, and the uniformity of conditions the cells are subject to during processing. Tools were developed for two areas of cell handling: cell fusion and cell transportation. Designs suitable for implementation in microscale lab-on-chip systems were evaluated. Tool development was predominantly experimental, assisted by numerical modelling. The experimental investigation concerned device fabrication and operational performance. A number of cell handling tool designs were built and tested. Coplanar electrodes are not commonly used in bovine NT and reports on their efficacy were not available. These electrodes, which are simple to fabricate, were tested to determine fusion rates achievable in comparison with those of the baseline procedure. A novel fusion device, the micropit, was designed to assist bovine cell pairing and electrofusion. It was initially uncertain whether this device was capable of achieving cell fusion. Tests were conducted; and cell fusion and micro-positioning were demonstrated, as was an increase in biological cell processing throughput. Many miniaturised lab-on-chip systems rely on cell transportation. One illustration in the baseline procedure is the on-chip transport of cells to the cell fusion device. Potential cell transport mechanisms for a miniature cloning system were evaluated by prototype construction and testing. These mechanisms included travelling wave dielectrophoresis and capillary fluid actuation. To facilitate automation of on-chip cell transportation, a low cost electrically isolated cell detection system was developed based on a DVD pick-up unit. Various obstacles that were encountered during the course of device construction are noted, as are the fabrication methods employed.

The fibroblast growth factor family of polypeptides currently consists of nine structurally-related members. Cloning of the mouse homologue of the latest reported member of the family, FGF-9, is described in this study. Mouse Fgf9 exhibits a high level of sequence conservation with the human, rat and Xenopus counterparts. Of note is the lack of a hydrophobic signal peptide at the N-terminus of the coding sequence. The protein, however, appeared to be secreted by producer cells since a significant quantity of the protein could be purified from the culture supernatant of transfected cells. Members of the FGF family are known to bind to cell surface tyrosine kinase receptors (FGFRs) to elicit a variety of physiological responses. These receptors themselves form a family of four structurally-related tyrosine kinases and each FGF member commonly has the ability to bind several members of the FGFR family. By using in vitro plate binding assays, FGF-9 is shown in this study to bind specifically to two FGFR members: FGFR2b and FGFR3c. To further study the potential functional role of FGF-9, its expression pattern in the mouse embryo was examined by both RNase protection and RNA in situ hybridization analyses. The transcript was detected in a variety of embryonic tissues: the germinal epithelium of the central nervous system, the mesonephric cords, the somites, the gut primordia and the developing eye and ear, suggesting that the gene may have multiple roles during development. In addition, the potential involvement of FGF-9 in mediating adult brain functions was examined by double RNA in situ hybridization analysis of the distribution of both Fgf9 and Fgfr3 transcripts in the adult mouse brain. Most apparent areas of co-localization are the olfactory bulb and cerebral cortex. The two transcripts are also shown to have distinct distribution patterns in the cerebellum.

Thesis (Ph. D.)--University of Hawaii at Manoa, 1995. / Includes bibliographical references (leaves 145-154). / Microfiche. / xiii, 159 leaves, bound ill., photos. 29 cm