Expression of Enterovirus 71 Structural Proteins and Formation of Virus-Like Particle in Insect Cells
| Autor: | Jen-Huang Huang, 黃振煌 |
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| Rok vydání: | 2003 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 91 Enterovirus 71 (EV71) is one of the 69 enterovirus serotypes within the picornavirus family. It has been a major etiological agent in the outbreaks of severe neurological diseases in children and has caused almost two hundred people death. The prevention and treatment, however, have been hindered owing to the lack of information on virus structure, functional roles of individual proteins, etc. The study of enterovirus-like particles (VLPs) could supplement the missing information and serve as a potential vaccine. The assembly of enterovirus, according to a model derived from poliovirus, relies on the cleavage of P1 polyprotein into individual structural proteins by protease 3CD. In this study, we constructed three recombinant baculoviruses, one expressing P1 polyprotein, one expressing 3CD protease and the other one expressing both P1 and 3CD. The co-expression of both P1 and 3CD by either co-infection or single infection resulted in the processing of the precursor protein P1 into VP1 in the cells. We also use in vitro proteolytic experiment to demonstrate the ability of 3CD for further discussion of assemble of VLP. The electron microscopy analysis revealed crystalline virus-like particle structures in the cells that appeared morphologically indistinguishable from authentic EV71. The characterization of protein expression and VLP formation are presented. The production and purification methods also are optimized for future vaccine application. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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