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dc.contributor.advisorMeiring, S. M.
dc.contributor.advisorDeckmyn, H.
dc.contributor.authorDe Bruin, Karen
dc.date.accessioned2015-08-05T13:55:56Z
dc.date.available2015-08-05T13:55:56Z
dc.date.issued2004-06
dc.identifier.urihttp://hdl.handle.net/11660/781
dc.description.abstractEnglish: The need to find new manners in which to combat cardiovascular disease and associated thrombotic complications, remains a high priority in industrialised countries. Even in third-world countries the implications and associated risks of these diseases are being felt more and more. The advent of the biotechnology era and employment of recombinant DNA techniques has brought about exponential advances in understanding the complex mechanisms of haemostasis, and is employed to find new ways to combat pathological thrombotic complications. The challenge is to harness the many tools and techniques produced by the ongoing biotechnology explosion, and apply them to elucidate questions still unanswered and explore areas still unknown. In this study it was illustrated that modern molecular biology techniques can be applied in many areas of thrombosis and haemostasis research. The display of cDNA libraries on the surfaces of filamentous bacteriophages was used in the search for novel antithrombotic compounds from a haematophagous insect Hippobosca rufipes. Phages displaying the cDNA libraries were panned against human a-thrombin and selected according to their binding affinity and inhibition ability. To illustrate the use of a Escherichia coli expression system, a domain of a enzyme was cloned, expressed, and the recombinant peptide isolated and refolded. ADAMTS-13 was recently identified as an important role player in the realm of von Willebrand factor activity, including primary haemostasis and pathological disorders. The second carboxy-terminal CUB domain of ADAMTS-13 was amplified from full-length cDNA, cloned into a expression vector system, and expressed as insoluble inclusion bodies in the cytoplasm of E. coli, from where it was isolated and refolded. In this study, molecular techniques were used in different phases of research into the specific activity and interactions of a particular component of the haemostatic system. This illustrated the marriage of biotechnology with fundamental medical research in an era of interdisciplinary sciences.en_ZA
dc.description.abstractAfrikaans: Die voortdurende soeke na nuwe maniere om kardiovaskulêre siektes die hoof te bied geniet hoë prioriteit in eerste-wêreld lande, terwyl die impak van hierdie siekte toestande meer en meer in die derde-wêreld gevoel word. Die aanvang en onlangse ontploffing van die biotegnologiese era het die toepassing van rekombinante DNA beginsels en tegnieke vandag alledaags gemaak. Mediese biotegnologie het reuse vooruitgang gebring in die diepte waarin die ingewikkelde wisselwerking van die komptenente betrokke in hemostase verstaan word, en word uiteindelik ingespan om ongewenste trombose te beveg. Die uitdaging is daarin geleë om al die nuwe tegnologie en tegnieke wat uit die biotegnologiese revolusie voortspring, in te span om onbeantwoorde vrae en raaisels op te los. Hierdie studie illustreer hoe moderne tegnieke in molekulêre biologie aangewend kan word in verskillende gebiede van hemostase en trombose navorsing. Die blootlegging van komplementêre DNA (cDNA)-biblioteke op die oppervlakte van filamenteuse bakteriofage was gebruik in die soeke om nuwe, unieke antistolmiddels uit die bloedsuiende perde-luisvlieg Hippobosca rufipes te isoleer. cDNA-blootleggende fage is getoets teen menslike a-trombien en verder geselekteer op grond van hul affiteits bindings- en inhibisievermoëns. Ten einde die werking van die Escherichia coli uitdrukkingssiteem te illustreer, is ‘n domein van ‘n ensiem gekloneer en uitgedruk, en die rekombinante peptied is geïsoleer en hervou. ADAMTS-13 is onlangs aangewys as ‘n belangrike rolspeler in die bepaling van von Willebrand-faktor aktiwiteit in beide primêre hemostase en siekte toestande. Die tweede CUB domein aan die karboksie-einde van ADAMTS-13 is vanaf volledige cDNA geamplifiseer, gekloneer in ‘n uitdrukkingsvektorsisteem, en uitgedruk in onoplosbare liggaampies in die E. coli sitoplasma, waarvandaan dit geïsoleer en hervou is. In hierdie studie is molekulêre tegnieke ingespan in verskillende navorsingsfases, in die poging om die spesifieke aktiwiteite en wisselwerkinge waarby ‘n bepaalde kompenent van die hemostase stelsel betrokke is, beter te verstaan. Dit dien as voorbeeld van effektiewe samewerking tussen biotegnologie en basiese mediese navorsing in hierdie opwindende tye van interdissiplinêre wetenskappe.
dc.language.isoenen_ZA
dc.publisherUniversity of the Free Stateen_ZA
dc.subjectBlood coagulation disordersen_ZA
dc.subjectHemostasisen_ZA
dc.subjectBlood -- Molecular aspectsen_ZA
dc.subjectDissertation (M.Med.Sc. (Haematology and Cell Biology))--University of the Free State, 2004en_ZA
dc.titleConstruction of cDNA libraries, and the selection and expression of proteins and peptides involved in haemostasisen_ZA
dc.typeDissertationen_ZA
dc.rights.holderUniversity of the Free Stateen_ZA


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