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Purification of Rice Stripe Virus

Authors: Gang Lu, Min Yao, Yijun Zhou and Xiaorong Tao, date: 03/20/2020, view: 152, 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 ...

Zebrafish Bacterial Infection Assay to Study Host-Pathogen Interactions

Authors: Faiza Basheer, Clifford Liongue and Alister C Ward, date: 03/05/2020, view: 274, Q&A: 0

[Abstract] The study of host-pathogen interactions has improved our understanding of both pathogenesis and the response of the host to infection, including both innate and adaptive responses. Neutrophils and macrophages represent the first line of innate host defense against any infection. The zebrafish is an ideal model to study the response of these cells ...

Transcervical Mouse Infections with Chlamydia trachomatis and Determination of Bacterial Burden

Authors: Karthika Rajeeve and Rajeeve Sivadasan, date: 02/05/2020, view: 341, Q&A: 0

[Abstract] Chlamydia trachomatis is an obligate human pathogen. It infects the genital tract of humans ascending into the fallopian tube, exacerbated by chronic pelvic pain, pelvic inflammatory disease, and fallopian tube scaring resulting in infertility and other malignancies. The major hurdle in controlling chlamydial spread is that the ...

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: 384, 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: 459, 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 ...

Study of Microbial Extracellular Vesicles: Separation by Density Gradients, Protection Assays and Labelling for Live Tracking

Authors: Carolina Coelho, Raghav Vij, Daniel Q. Smith, Nathan R. Brady, Anne Hamacher-Brady and Arturo Casadevall, date: 01/20/2020, view: 495, Q&A: 0

[Abstract] Extracellular vesicles (EVs) are produced by all domains of life including Bacteria, Archaea and Eukarya. EVs are critical for cellular physiology and contain varied cargo: virulence factors, cell wall remodeling enzymes, extracellular matrix components and even nucleic acids and metabolites. While various protocols for isolating EVs have been ...

A Microbial Bioassay for Direct Contact Assessment of Soil Toxicity Based on Oxygen Consumption of Sulfur Oxidizing Bacteria

Authors: Ebenezer Ashun, Umair Ali Toor, Heonseop Eom and Sang-Eun Oh, date: 01/05/2020, view: 441, Q&A: 0

[Abstract] A new direct contact assessment of soil toxicity using sulfur oxidizing bacteria (SOB) is proposed for analyzing the toxicity of soils. The proposed method is based on the ability of SOB to oxidize elemental sulfur to sulfuric acid in the presence of oxygen. Since sulfate ions are produced from sulfur by SOB oxidation activity, changes in ...

Propagation and Purification of Chlamydia trachomatis Serovar L2 Transformants and Mutants

Authors: Robert Faris and Mary M. Weber, date: 12/20/2019, view: 560, Q&A: 0

[Abstract] Chlamydia trachomatis (C.t.) is an obligate intracellular pathogen that cannot be cultured axenically and must be propagated within eukaryotic host cells. There are at least 15 distinct chlamydial serovariants that belong to 2 major biovars commonly referred to as trachoma and lymphogranuloma venereum (LGV). The invasive ...

Lysosome Targeting RedGreen-assay: Selective Autophagy Sensing Assay for Mammalian Cells

Authors: Victoria Riccio, Miluska Vissa, Angus McQuibban and Peter Kijun Kim, date: 12/20/2019, view: 929, Q&A: 0

[Abstract] The process of autophagy is an essential cellular mechanism, required to maintain general cell health through the removal of dysfunctional organelles, such as the ER, peroxisomes and mitochondria, as well as protein aggregates, and bacteria. Autophagy is an extremely dynamic process, and tools are constantly being developed to study the various ...

Preparation of Red Palm Weevil Rhynchophorus Ferrugineus (Olivier) (Coleoptera: Dryophthoridae) Germ-free Larvae for Host-gut Microbes Interaction Studies

Authors: Abrar Muhammad, Prosper Habineza, Youming Hou and Zhanghong Shi, date: 12/20/2019, view: 650, Q&A: 0

[Abstract] Red palm weevil (RPW), Rhynchophorus ferrugineus Olivier, is a devastating pest of palm trees worldwide. RPW gut is colonized by diverse bacterial species which profoundly influence host development and nutritional metabolism. However, the molecular mechanisms behind the interactions between RPW and its gut microbiota remain mostly ...

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