Human protein C (HPC) is a vitamin-K dependent plasma glycoprotein which is one of the major components regulating anticoagulation. HPC injection is a promising therapy for several diseases but a heterologous production system would be preferred over purifying HPC from human plasma because of its low concentration (4-5 $ mu$g/ml). A cDNA clone coding for HPC was inserted downstream of the CaMV 35S promoter and of a dimer of the CaMV 35S promoter. Tobacco plants were transformed using Agrobacterium and a binary vector strategy. Kanamycin resistant plants were regenerated and enzyme linked immunosorbent assay determined that HPC, in crude plant extracts, accounted for up to 0.03% of plant soluble proteins. HPC was found to be expressed by R$ sb1$ seedlings suggesting successful integration of the T-DNA into plant genome. A partial protein purification system was developed in order to enrich the protein mixture for HPC. HPC was found to bind tightly at pH 6.0 to Fast Flow Q Sepharose resin.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.55455 |
| Date | January 1994 |
| Creators | Piché, Christian. |
| Contributors | Fortin, Marc G. (advisor) |
| Publisher | McGill University |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Format | application/pdf |
| Coverage | Master of Science (Department of Plant Science.) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 001538786, proquestno: AAIMM19699, Theses scanned by UMI/ProQuest. |
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