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Purification and Proteomic Analysis of Alphavirus Particles from Sindbis Virus Grown in Mammalian and Insect Cells

Authors: Raquel Hernandez, Trevor Glaros, Gabrielle Rizzo and Davis F. Ferreira, date: 05/20/2019, view: 309, Q&A: 0

[Abstract] Current mass spectrometry (MS) methods and new instrumentation now allow for more accurate identification of proteins in low abundance than previous protein fractionation and identification methods. It was of interest if this method could serve to define the virus proteome of a membrane-containing virus. To evaluate the efficacy of mass spec to ...

Viral Chromosome Conformation Capture (V3C) Assays for Identifying Trans-interaction Sites between Lytic Viruses and the Cellular Genome

Authors: Kinjal Majumder, Maria Boftsi and David J Pintel, date: 03/20/2019, view: 543, Q&A: 0

[Abstract] The folding mechanisms of the mammalian genome package our genetic material into the nucleus, and in doing so, dictate its appropriate replication and expression. Chromosome conformation capture technology has enabled the dissection of the folding principles of the cellular genome. This has led to a better understanding of the role played by ...

Quantitation of Regulatory Activity for the Complement Alternative Pathway Using an Adaptation of the AP₅₀ in vitro Assay

Authors: Sheila Cabezas Falcon, David L Gordon and Jillian M Carr, date: 12/20/2018, view: 835, Q&A: 0

[Abstract] Complement pathways function to identify and remove pathogens and infected cells. There are three complement pathways: the classical, lectin and alternative pathway (AP). While all pathways are activated following pathogen stimuli, the AP is constitutively active and tightly controlled by activators (e.g., Factor B, Factor D) and negative ...

Quantification of Infectious Sendai Virus Using Plaque Assay

Authors: Narihito Tatsumoto, Takamasa Miyauchi, Moshe Arditi and Michifumi Yamashita, date: 11/05/2018, view: 1293, Q&A: 0

[Abstract] Sendai virus (SeV) is an enveloped, single-stranded RNA virus of the family Paramyxoviridae. SeV is a useful tool to study its infectious pathomechanism in immunology and the pathomechanism of a murine model of IgA nephropathy. Virus quantification is essential not only to determine the original viral titers for an appropriate ...

Rice Ragged Stunt Virus Propagation and Infection on Rice Plants

Authors: Chao Zhang, Chaonan Shi, Dong Chen and Jianguo Wu, date: 10/20/2018, view: 1128, Q&A: 0

[Abstract] Virus inoculation is a basic experimental procedure to evaluate the resistance of a rice variety or a transgenic material upon virus infection. We recently demonstrated that Rice Ragged Stunt Virus (RRSV), an oryzavirus that is transmitted by brown planthopper (BPH), can suppress jasmonic acid-mediated antiviral defense through ...

HIV_GKO: A Tool to Assess HIV-1 Latency Reversal Agents in Human Primary CD4⁺ T Cells

Authors: Emilie Battivelli and Eric Verdin, date: 10/20/2018, view: 1387, Q&A: 0

[Abstract] While able to suppress HIV replication in HIV infected individuals, combination antiretroviral therapy (ART) fails to eliminate viral latent reservoir, which consists in integrated transcriptional silenced HIV provirus. So far, identification of latently-infected cells has relied on activating cells to induce expression of HIV proteins which can ...

Retroviral Capsid Core Stability Assay

Authors: Tyler Milston Renner, Kasandra Bélanger and Marc-André Langlois, date: 09/20/2018, view: 1332, Q&A: 0

[Abstract] Structural stability of the capsid core is a critical parameter for the productive infection of a cell by a retrovirus. Compromised stability can lead to premature core disassembly, exposure of replication intermediates to cytosolic nucleic acid sensors that can trigger innate antiviral responses, and failure to integrate the proviral genome into ...

Liposome Flotation Assay for Studying Interactions Between Rubella Virus Particles and Lipid Membranes

Authors: Kyoko Saito, Noriyuki Otsuki, Makoto Takeda and Kentaro Hanada, date: 08/20/2018, view: 1920, Q&A: 0

[Abstract] Rubella virus (RuV) is an enveloped, positive-sense single-stranded RNA virus that is pathogenic to humans. RuV binds to the target cell via the viral envelope protein E1, but the specific receptor molecules on the target cell are yet to be fully elucidated. Here, we describe a protocol for liposome flotation assay to study direct interactions ...

Random Insertional Mutagenesis of a Serotype 2 Dengue Virus Clone

Authors: Jeffrey W. Perry and Andrew W. Tai, date: 08/20/2018, view: 1269, 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 ...

BMV Propagation, Extraction and Purification Using Chromatographic Methods

Authors: Aleksander Strugała, Paulina Bierwagen, Jakub Dalibor Rybka, Michał Giersig, Marek Figlerowicz and Anna Urbanowicz, date: 07/20/2018, view: 1482, Q&A: 0

[Abstract] Brome mosaic virus (BMV) is a well-known plant virus representing single-stranded RNA (ssRNA) positive-sense viruses. It has been widely used as a model in multiple studies concerning plant virus biology, epidemiology and the application of viral capsids in nanotechnology. Herein, we describe a method for BMV purification based on ion-exchange and ...

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