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Expression of human tissue plasminogen activator using the Bombyx mori nuclear polyhedrosis virus system

Authors Tang, Rocky P.N.
Issue Date 1994
Summary In this study, we utilized the BmNPV to express active human tissue plasminogen activator, a potent fibrinolytic agent, in BmN cells and the silkworm Bombyx mori. Two expression vectors, one containing the original t-PA cDNA and the other a mutated t-PA cDNA, were constructed. The BmLSP signal peptide was used to replace the original t-PA signal peptide, and a short sequence containing 7 bases, TATAAAT, was added immediately before the signal peptide. Recombinant viruses were generated from these vectors, and were used to infect BmN cells. Media from a titne course experiment of 5 days were studied. The radial caseinolysis assay was employed to check the activities of the expressed products. This study gave positive indication that human t-PA could be expressed by the BmNPV system. It seemed that the mutation introduced into the t-PA gene did enhance the level of expression and secretion to a certain extent. However, more work need to be done on the characterization and the purification of the expressed products.
Note Thesis (M.Phil.)--Hong Kong University of Science and Technology, 1994
Language English
Format Thesis
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