Abstract
Chikungunya virus (CHIKV) is the etiologic agent of Chikungunya fever and has emerged in many countries over the past decade. There are no effective drugs for controlling the disease. A bicistronic baculovirus expression system was utilized to co-express CHIKV 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 is mediated by the CHIKV E1 allowing it to identify chemicals that can prevent syncytium formation. The compounds characterized by this method could be anti-CHIKV drugs.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Griffin DE (2007) Alphaviruses. In: Knipe DM, Howley PM (eds) Fields virology, 5th edn. Lippincott-Williams & Wilkins, Philadelphia, pp 1023–1068
Solignat M, Gay B, Higgs S, Briant L, Devaux C (2009) Replication cycle of Chikungunya: a re-emerging arbovirus. Virology 393:183–197
Powers AM, Logue CH (2007) Changing patterns of chikungunya virus: re-emergence of a zoonotic arbovirus. J Gen Virol 88:2363–2377
Ross RW (1956) A laboratory technique for studying the insect transmission of animal viruses, employing a bat-wing membrane, demonstrated with two African viruses. J Hyg (Lond) 54:192–200
Rezza G, Nicoletti L, Angelini R, Romi R, Finarelli AC, Panning M, Cordioli P, Fortuna C, Boros S, Magurano F (2007) Infection with Chikungunya virus in Italy: an outbreak in a temperate region. Lancet 370:1840–1846
Unger T, Peleg Y (2012) Recombinant protein expression in the baculovirus-infected insect cell system. Methods Mol Biol 800:187–199
Chen WS, Villaflores OB, Lu CF, Wu HI, Chen YJ, Teng CY, Chang YC, Chang SL, Wu TY (2012) Functional expression of rat neuroligin-1 extracellular fragment by a bi-cistronic baculovirus expression vector. Protein Expr Purif 81:18–24
Wu TY, Chen YJ, Teng CY, Chen WS, Villaflores O (2012) A bi-cistronic baculovirus expression vector for improved recombinant protein production. Bioeng Bugs 3:129–132
Kuo SC, Chen YJ, Wang YM, Kuo MD, Jinn TR, Chen WS, Chang YC, Tung KL, Wu TY, Lo SJ (2011) Cell-based analysis of Chikungunya virus membrane fusion using baculovirus-expression vectors. J Virol Methods 175:206–215
Metz SW, Geertsema C, Martina BE, Andrade P, Heldens JG, van Oers MM, Goldbach RW, Vlak JM, Pijlman GP (2011) Functional processing and secretion of Chikungunya virus E1 and E2 glycoproteins in insect cells. Virol J 8:353
Kuo SC, Chen YJ, Wang YM, Tsui PY, Kuo MD, Wu TY, Lo SJ (2012) Cell-based analysis of Chikungunya virus E1 protein in membrane fusion. J Biomed Sci 19:44
Woolaway KE, Lazaridis K, Belsham GJ, Carter MJ, Roberts LO (2001) The 5′ untranslated region of Rhopalosiphum padi virus contains an internal ribosome entry site which functions efficiently in mammalian, plant, and insect translation systems. J Virol 75:10244–10249
Chen YJ, Chen WS, Wu TY (2005) Development of a bi-cistronic baculovirus expression vector by the Rhopalosiphum padi virus 5′ internal ribosome entry site. Biochem Biophys Res Commun 335:616–623
Blissard GW, Wenz JR (1992) Baculovirus gp64 envelope glycoprotein is sufficient to mediate pH-dependent membrane fusion. J Virol 66:6829–35
Maruniak JE, Garcia-Canedo A, Rodrigues JJ (1994) Cell lines used for the selection of recombinant baculovirus. In Vitro Cell Dev Biol Anim 30:283–286
Acknowledgement
This research was supported by the Ministry of Science and Technology (NSC103-2321-B-033-001‚105-2321-B-033-001 & NSC 102-2632-M-033-001-MY3).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Kuo, SC., Teng, CY., Ho, YJ., Chen, YJ., Wu, TY. (2016). Using Bicistronic Baculovirus Expression Vector System to Screen the Compounds That Interfere with the Infection of Chikungunya Virus. In: Chu, J., Ang, S. (eds) Chikungunya Virus. Methods in Molecular Biology, vol 1426. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3618-2_24
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3618-2_24
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3616-8
Online ISBN: 978-1-4939-3618-2
eBook Packages: Springer Protocols