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Construction of Recombinant Baculoviruses Expressing Structural Proteins of Chikungunya Virus

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...

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Main Author: Choo, Chee Wei
Format: Thesis
Language:English
Published: Universiti Malaysia Sarawak (UNIMAS) 2019
Subjects:
Q Science (General)
QR Microbiology
Online Access:http://ir.unimas.my/id/eprint/27226/3/choo%20chee%20wei%20ft.pdf
http://ir.unimas.my/id/eprint/27226/
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spelling my.unimas.ir.272262024-08-19T03:05:32Z http://ir.unimas.my/id/eprint/27226/ Construction of Recombinant Baculoviruses Expressing Structural Proteins of Chikungunya Virus Choo, Chee Wei Q Science (General) QR Microbiology 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. Universiti Malaysia Sarawak (UNIMAS) 2019-10-13 Thesis NonPeerReviewed text en http://ir.unimas.my/id/eprint/27226/3/choo%20chee%20wei%20ft.pdf Choo, Chee Wei (2019) Construction of Recombinant Baculoviruses Expressing Structural Proteins of Chikungunya Virus. Masters thesis, Universiti Malaysia Sarawak (UNIMAS).
institution Universiti Malaysia Sarawak
building Centre for Academic Information Services (CAIS)
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaysia Sarawak
content_source UNIMAS Institutional Repository
url_provider http://ir.unimas.my/
language English
topic Q Science (General)
QR Microbiology
spellingShingle Q Science (General)
QR Microbiology
Choo, Chee Wei
Construction of Recombinant Baculoviruses Expressing Structural Proteins of Chikungunya Virus
description 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.
format Thesis
author Choo, Chee Wei
author_facet Choo, Chee Wei
author_sort Choo, Chee Wei
title Construction of Recombinant Baculoviruses Expressing Structural Proteins of Chikungunya Virus
title_short Construction of Recombinant Baculoviruses Expressing Structural Proteins of Chikungunya Virus
title_full Construction of Recombinant Baculoviruses Expressing Structural Proteins of Chikungunya Virus
title_fullStr Construction of Recombinant Baculoviruses Expressing Structural Proteins of Chikungunya Virus
title_full_unstemmed Construction of Recombinant Baculoviruses Expressing Structural Proteins of Chikungunya Virus
title_sort construction of recombinant baculoviruses expressing structural proteins of chikungunya virus
publisher Universiti Malaysia Sarawak (UNIMAS)
publishDate 2019
url http://ir.unimas.my/id/eprint/27226/3/choo%20chee%20wei%20ft.pdf
http://ir.unimas.my/id/eprint/27226/
_version_ 1808981474104311808
score 13.211869

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