Choo, Chee Wei (2019) Construction of Recombinant Baculoviruses Expressing Structural Proteins of Chikungunya Virus. Masters thesis, Universiti Malaysia Sarawak (UNIMAS).
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Abstract
Chikungunya virus (CHIKV) is a reemerging human pathogen that causes a debilitating arthritic-like disease in humans. Although numerous strategies have been adopted to combat CHIKV infection, transmission of CHIKV continues to increase globally. The most effective measure to reduce the CHIKV burden is the development of a vaccine. However, there is no licensed CHIKV vaccine available currently for human use. CHIKV VLPs have the potential to be used as a prophylactic vaccine and currently there are several CHIKV VLPs candidates (generated via mammalian expression system) being evaluated for human use in Phase I clinical trials. Accordingly, this study was proposed to construct CHIKV recombinant structural proteins which could be potentially used as a source of CHIK VLPs using an insect cell-baculovirus expression system and to test the antigenicity of the expressed structural proteins against known CHIKV patient’s sera. The selected gene region encoding CHIKV structural polyprotein (E3-E2-6K-EctoE1) was amplified as a BamHI-HindIII fragment and cloned into bac-to-bac® baculovirus expression vector. The resulting construct was used to transfected Sf9 cells to generate a recombinant baculovirus (AcMNPV-E3E26KEctoE1). Western blot analysis of the expressed recombinant protein, confirmed the presence of a 120 kDa expected size product. Western blot results demonstrated that the expressed CHIKV proteins were detected by sera from patients previously exposed to CHIKV infection. CHIKV positive patient sera also detected a lower than expected recombinant proteins of 38 and 39 kDa respectively in the western blot analysis. These may possibly indicate incomplete or unassembled component proteins of the VLP.
| Item Type: | Thesis (Masters) |
|---|---|
| Additional Information: | Thesis (MSc.) - Universiti Malaysia Sarawak , 2019. |
| Uncontrolled Keywords: | Chikungunya virus (CHIKV), Virus-like particles (VLPs), insect cells- baculovirus expression system, bac-to-bac® baculovirus expression system, unimas, university, universiti, Borneo, Malaysia, Sarawak, Kuching, Samarahan, ipta, education, research, Universiti Malaysia Sarawak. |
| Subjects: | Q Science > Q Science (General) Q Science > QR Microbiology |
| Divisions: | Academic Faculties, Institutes and Centres > Institute of Health and Community Medicine Faculties, Institutes, Centres > Institute of Health and Community Medicine |
| Depositing User: | CHOO CHEE WEI |
| Date Deposited: | 04 Oct 2019 01:12 |
| Last Modified: | 19 Aug 2024 03:05 |
| URI: | http://ir.unimas.my/id/eprint/27226 |
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