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:	2019
Subjects:	Q Science (General) QR Microbiology
Online Access:	http://ir.unimas.my/id/eprint/27226/2/choo%20chee%20wei%20ft.pdf
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id	my-unimas-ir.27226
record_format	uketd_dc
spelling	my-unimas-ir.272262024-07-15T08:56:54Z Construction of Recombinant Baculoviruses Expressing Structural Proteins of Chikungunya Virus 2019-10-13 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 Thesis http://ir.unimas.my/id/eprint/27226/ http://ir.unimas.my/id/eprint/27226/2/choo%20chee%20wei%20ft.pdf text en validuser masters Universiti Malaysia Sarawak (UNIMAS) Institute of Health and Community Medicine
institution	Universiti Malaysia Sarawak
collection	UNIMAS Institutional Repository
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
qualification_level	Master's degree
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
granting_institution	Universiti Malaysia Sarawak (UNIMAS)
granting_department	Institute of Health and Community Medicine
publishDate	2019
url	http://ir.unimas.my/id/eprint/27226/2/choo%20chee%20wei%20ft.pdf
_version_	1804888398689730560

Construction of Recombinant Baculoviruses Expressing Structural Proteins of Chikungunya Virus

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