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Purification of Rice Stripe Virus

Authors: Gang Lu, Min Yao, Yijun Zhou and Xiaorong Tao, date: 03/20/2020, view: 203, Q&A: 0

[Abstract] Although many spherical and rod-shaped plant virus purification protocols are now available, only a few protocols on filamentous plant virus purification have been published. Here, we report a protocol for large-scale purification of Rice stripe virus (RSV) from RSV-infected rice tissues. RSV virions with high infectivity were first precipitated ...

Expression and Purification of Adeno-associated Virus Virus-like Particles in a Baculovirus System and AAVR Ectodomain Constructs in E. coli

Authors: Nancy Meyer, Omar Davulcu, Qing Xie, Mark Silveria, Grant M. Zane, Edward Large and Michael S. Chapman, date: 02/05/2020, view: 403, Q&A: 0

[Abstract] Adeno-associated virus (AAV) is a promising gene therapy vector and the biophysical characterization of its interactions with host proteins is a critical foundation for engineering tissue targeting and immune escape. Presented here are protocols for the production of: (a) the outer protein shells (virus-like particles or VLPs) for serotype 2 ...

Quantifying HIV-1-Mediated Gut CD4⁺ T Cell Death in the Lamina Propria Aggregate Culture (LPAC) Model

Authors: Stephanie M. Dillon, Kejun Guo, Moriah J. Castleman, Mario L. Santiago and Cara C. Wilson, date: 01/20/2020, view: 471, Q&A: 0

[Abstract] Gut CD4 T cells are major targets of HIV-1 and are massively depleted early during infection. To better understand the mechanisms governing HIV-1-mediated CD4 T cell death, we developed the physiologically-relevant Lamina Propria Aggregate Culture (LPAC) model. The LPAC model is ideal for studying CD4 T cell death induced by clinically-relevant ...

Herpes Simplex Virus Type 1 Propagation, Titration and Single-step Growth Curves

Authors: Linda Grosche, Katinka Döhner, Alexandra Düthorn, Ana Hickford-Martinez, Alexander Steinkasserer and Beate Sodeik, date: 12/05/2019, view: 1091, Q&A: 0

[Abstract] Given the endemic seroprevalence of herpes simplex viruses (HSV), its associated human diseases, and the emergence of acyclovir-resistant strains, there is a continuous need for better antiviral therapies. Towards this aim, identifying mechanistic details of how HSV-1 manipulates infected cells, how it modulates the immune responses, and how it ...

Production of Quantum Dots-containing Influenza Virus Particles for Studying Viral Uncoating Processes

Authors: Chong Qin, Xiaowei Zhang and Zongqiang Cui, date: 09/20/2019, view: 868, Q&A: 0

[Abstract] The genome of influenza A virus (IAV) comprises eight pinlike genomic segments called vRNPs enclosed in viral capsid. During infection, uncoating is the key step for viral replication and represents an antiviral therapeutic target, but it is difficult to observe the transient and dynamic event in detail. Here, we report a protocol for production ...

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: 913, 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 ...

Fluorescence Microscopy Assay to Measure HIV-1 Capsid Uncoating Kinetics in vitro

Authors: Chantal L. Márquez, Derrick Lau, James Walsh, K. M. Rifat Faysal, Michael W. Parker, Stuart G. Turville and Till Böcking, date: 07/05/2019, view: 1436, Q&A: 0

[Abstract] The stability of the HIV-1 capsid and the spatiotemporal control of its disassembly, a process called uncoating, need to be finely tuned for infection to proceed. Biochemical methods for measuring capsid lattice disassembly in bulk are unable to resolve intermediates in the uncoating reaction. We have developed a single-particle fluorescence ...

In vitro Infection of Primary Human Monocytes with HIV-1

Authors: Patrick O’Connell, Yong-Hui Zheng, Andrea Amalfitano and Yasser A. Aldhamen, date: 07/05/2019, view: 1269, Q&A: 0

[Abstract] Monocyte infection by HIV-1 is an important component of chronic HIV pathogenesis. Following infection by HIV-1, monocytes are able to cross the blood brain barrier and set up a viral reservoir in the central nervous system. Additionally, in the setting of chronic HIV-1 infection, monocytes can become activated either directly through HIV-1 ...

Analysis of Functional Virus-generated PAMP RNAs Using IFNα/β ELISA Assay

Authors: Margit Mutso, Xiang Liu, Andres Merits and Suresh Mahalingam, date: 06/20/2019, view: 991, Q&A: 0

[Abstract] Virus-generated PAMP RNAs are key factors that activate host immune response. The PAMP RNAs are therefore usually closely related with viral disease pathogenesis. Quantitative real time PCR is a conventional method to assess RNA. However, it cannot be used for detecting short dsRNAs generated by viral replicase. This protocol was established to ...

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: 1841, 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 ...

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