Viruses

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    Protocols in Current Issue
    Image-based Quantification of Direct Cell-to-cell Transmission of Bovine Viral Diarrhea Virus
    Authors:  Fernando Merwaiss and Diego E. Alvarez, date: 08/05/2019, view: 550, Q&A: 0
    [Abstract] Different viruses rely on direct cell-to-cell transmission to propagate infection within the infected host. Measuring this mode of transmission in cultured cells is often complicated by the contribution of cell free viruses to spread, and the difficulty to distinguish between primary infected cells that produce the virus and neighboring cells that ...
    Production and Purification of Dengue Virus-like Particles from COS-1 Cells
    Authors:  Jedhan Ucat Galula, Gwong-Jen J. Chang and Day-Yu Chao, date: 06/20/2019, view: 1192, Q&A: 0
    [Abstract] Non-infectious virus-like particles (VLPs) containing dengue virus (DENV) pre-membrane (prM) and envelope (E) proteins have been demonstrated to be highly immunogenic and can be used as a potential vaccine candidate as well as a tool for serodiagnostic assays. Successful application of VLPs requires abundant, and high-purity production methods. ...
    Production, Titration and Imaging of Zika Virus in Mammalian Cells
    Authors:  Wesley Freppel, Clément Mazeaud and Laurent Chatel-Chaix, date: 12/20/2018, view: 2510, Q&A: 0
    [Abstract] Since the outbreak of Zika virus (ZIKV) in Latin America and the US in 2016, this flavivirus has emerged as a major threat for public health. Indeed, it is now clear that ZIKV is vertically transmitted from the infected mother to the fetus and this may lead to severe neurological development defects including (but not restricted to) neonate ...
    In vitro Membrane Interaction and Liposome Fusion Assays Using Recombinant Hepatitis C Virus Envelope Protein E2
    [Abstract] In order to study the mechanism underlying the Hepatitis C Virus (HCV) fusion process we have performed assays using phospholipid liposomes and a truncated form of E2 protein, E2661 (amino acids 384-661 of the HCV polyprotein) lacking the transmembrane region. E2661 has been previously generated by using the baculovirus ...
    Detection and Differentiation of Multiple Viral RNAs Using Branched DNA FISH Coupled to Confocal Microscopy and Flow Cytometry
    Authors:  Nicholas van Buuren and Karla Kirkegaard, date: 10/20/2018, view: 1934, Q&A: 0
    [Abstract] Due to the exceptionally high mutation rates of RNA-dependent RNA polymerases, infectious RNA viruses generate extensive sequence diversity, leading to some of the lowest barriers to the development of antiviral drug resistance in the microbial world. We have previously discovered that higher barriers to the development of drug resistance can be ...
    Random Insertional Mutagenesis of a Serotype 2 Dengue Virus Clone
    Authors:  Jeffrey W. Perry and Andrew W. Tai, date: 08/20/2018, view: 1962, Q&A: 0
    [Abstract] Protein tagging is a powerful method of investigating protein function. However, modifying positive-strand RNA virus proteins in the context of viral infection can be particularly difficult as their compact genomes and multifunctional proteins mean even small changes can inactivate or attenuate the virus. Although targeted approaches to ...
    HCV Reporter System (Viral Infection-Activated Split-Intein-Mediated Reporter System) for Testing Virus Cell-to-cell Transmission ex-vivo
    Authors:  Fanfan Zhao, Ting Zhao, Libin Deng, Dawei Lv, Xiaolong Zhang, Xiaoyu Pan, Jun Xu and Gang Long, date: 08/05/2018, view: 2157, Q&A: 0
    [Abstract] Hepatitis C virus (HCV) spread involves two distinct entry pathways: cell-free transmission and cell-to-cell transmission. Cell-to-cell transmission is not only an efficient way for viruses to spread but also an effective method for escaping neutralizing antibodies. We adapted the viral infection-activated split-intein-mediated reporter system ...
    Virucidal and Neutralizing Activity Tests for Antiviral Substances and Antibodies
    Authors:  Chie Aoki-Utsubo, Ming Chen and Hak Hotta, date: 05/20/2018, view: 3927, Q&A: 0
    [Abstract] In a narrow definition, virucidal activity represents the activity by which to interact with and physically disrupt viral particles. In a broad definition, it includes the activity by which to functionally inhibit (neutralize) viral infectivity without apparent morphological alterations of the viral particles. The viral infectivity can be measured ...
    Time-of-addition and Temperature-shift Assays to Determine Particular Step(s) in the Viral Life Cycle that is Blocked by Antiviral Substance(s)
    Authors:  Chie Aoki-Utsubo, Ming Chen and Hak Hotta, date: 05/05/2018, view: 2924, Q&A: 0
    [Abstract] Viruses infect their host cells to produce progeny virus particles through the sequential steps of the viral life cycle, such as viral attachment, entry, penetration and post-entry events. This protocol describes time-of-addition and temperature-shift assays that are employed to explore which step(s) in the viral life cycle is blocked by an ...
    Lentiviral Knockdown of Transcription Factor STAT1 in Peromyscus leucopus to Assess Its Role in the Restriction of Tick-borne Flaviviruses
    Authors:  Adaeze O. Izuogu and R. Travis Taylor, date: 12/05/2017, view: 5014, Q&A: 0
    [Abstract] Cellular infection with tick-borne flaviviruses (TBFVs) results in activation of the interferon (IFN) signaling pathway and subsequent upregulation of numerous genes termed IFN stimulated genes (ISGs) (Schoggins et al., 2011). Many ISGs function to prevent virus pathogenesis by acting in a broad or specific manner through protein-protein ...



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