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.
| Identifer | oai:union.ndltd.org:netd.ac.za/oai:union.ndltd.org:ufs/oai:etd.uovs.ac.za:etd-09292005-153019 |
| Date | 29 September 2005 |
| Creators | De Bruin, Karen |
| Contributors | Dr SM Meiring, Prof H Deckmyn |
| Publisher | University of the Free State |
| Source Sets | South African National ETD Portal |
| Language | en-uk |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.uovs.ac.za//theses/available/etd-09292005-153019/restricted/ |
| Rights | unrestricted, I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to University Free State or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
Page generated in 0.0014 seconds