Abstract
Cryo-electron microscopy (cryo-EM) is a valuable tool used to study the structures of icosahedral viruses without having to resort to crystallization. During the last few decades, significant progress has been made where virus structures previously resolved only to low resolution have now breached the sub-nanometer threshold. Critical to such excellent results are the acquisition of highly purified virus samples and well-frozen samples in vitreous ice. With the virus particles locked in their native conformations, cryo-EM together with single-particle analysis can then be deployed to study the structures of the viruses in their fully hydrated states.
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References
Chang J, Liu X, Rochat RH et al (2012) Reconstructing virus structures from nanometer to near-atomic resolutions with cryo-electron microscopy and tomography. Adv Exp Med Biol 726:49–90
Lindenbach BD, Rice CM (2001) Flaviviridae: the viruses and their replication. In: Knipe DM, Howley PM (eds) Fields virology, vol II, 4th edn. Lippincott Williams and Wilkins, Philadelphia, PA, pp 991–1042
Zhang Y, Corver J, Chipman PR et al (2003) Structures of immature flavivirus particles. EMBO J 22(11):2604–2613
Zhang Y, Kaufmann B, Chipman PR et al (2007) Structure of immature West Nile virus. J Virol 81(11):6141–6145
Kuhn RJ, Zhang W, Rossmann MG et al (2002) Structure of dengue virus: implications for flavivirus organization, maturation and fusion. Cell 108:717–725
Stadler K, Allison SL, Schalich J et al (1997) Proteolytic activation of tick-borne encephalitis virus by furin. J Virol 71(11):8475–8481
Yu IM, Zhang W, Holdaway HA et al (2008) Structure of the immature dengue virus at low pH primes proteolytic maturation. Science 319(5871):1834–1837
Ferreira GP, Trindade GS, Vilela JMC et al (2008) Climbing the steps of viral atomic force microscopy: visualisation of dengue virus particles. J Microsc 231(Pt1):180–185
Junjhon J, Edwards TJ, Utaipat U et al (2010) Influence of pr-M cleavage on the heterogeneity of extracellular dengue virus particles. J Virol 84(16):8353–8358
Iancu CV, Tivol WF, Schooler JB et al (2006) Electron cryotomography sample preparation using the Vitrobot. Nat Protoc 1(6):2813–2891
Dobro MJ, Melanson LA, Jensen GJ et al (2010) Plunge freezing for electron cryomicroscopy. Methods Enzymol 481:63–82
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Tan, J.L., Lok, S.M. (2014). Dengue Virus Purification and Sample Preparation for Cryo-Electron Microscopy. In: Padmanabhan, R., Vasudevan, S. (eds) Dengue. Methods in Molecular Biology, vol 1138. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0348-1_4
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DOI: https://doi.org/10.1007/978-1-4939-0348-1_4
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