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Cell-based analysis of Chikungunya virus membrane fusion using baculovirus-expression vectors.
- Source :
-
Journal of virological methods [J Virol Methods] 2011 Aug; Vol. 175 (2), pp. 206-15. Date of Electronic Publication: 2011 May 17. - Publication Year :
- 2011
-
Abstract
- Chikungunya virus infection has emerged in many countries over the past decade. There are no effective drugs for controlling the disease. To develop cell-based system for screening anti-virus drugs, a bi-cistronic baculovirus expression system was utilized to co-express viral structural proteins C (capsid), E2 and E1 and the enhanced green fluorescence protein (EGFP) in Spodoptera frugiperda insect cells (Sf21). The EGFP-positive Sf21 cells fused with each other and with uninfected cells to form a syncytium, allowing characterization of cholesterol and low pH requirements for syncytium formation. Western blot analysis showed three structural proteins were expressed in baculovirus infected cells. The structural proteins of Chikungunya virus that is required for cell fusion was determined with various recombinant baculoviruses bearing different lengths of the viral structural protein genes. Protein E1 was required for cell fusion and indicating that Chikungunya viral membrane fusion was a class II membrane fusion. It was also demonstrated that the heterologous expression of alphavirus monomeric E1 can induce insect cell fusions. Furthermore, this cell-based system provides a model for studying class II viral membrane fusion.<br /> (Copyright © 2011 Elsevier B.V. All rights reserved.)
- Subjects :
- Animals
Cell Culture Techniques
Cell Fusion
Cell Line
Chikungunya virus genetics
Genes, Reporter
Green Fluorescent Proteins biosynthesis
Green Fluorescent Proteins genetics
Recombinant Fusion Proteins biosynthesis
Recombinant Fusion Proteins genetics
Spodoptera
Viral Structural Proteins genetics
Baculoviridae genetics
Chikungunya virus physiology
Gene Expression
Viral Structural Proteins biosynthesis
Virus Internalization
Subjects
Details
- Language :
- English
- ISSN :
- 1879-0984
- Volume :
- 175
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Journal of virological methods
- Publication Type :
- Academic Journal
- Accession number :
- 21619896
- Full Text :
- https://doi.org/10.1016/j.jviromet.2011.05.015