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Detection of in vivo Protein Interactions in All Bacterial Compartments by Förster Resonance Energy Transfer with the Superfolder mTurquoise2 ox-mNeongreen FRET Pair

Authors: Nils Y. Meiresonne, Elisa Consoli, Laureen M.Y. Mertens and Tanneke den Blaauwen, date: 12/05/2019, view: 2012, Q&A: 0

[Abstract] This protocol was developed to qualitatively and quantitatively detect protein-protein interactions in all compartments of Escherichia coli by Förster Resonance Energy Transfer (FRET) using the Superfolder mTurquoise2 ox-mNeonGreen FRET pair (sfTq2^ox-mNG). This FRET pair has more than twice the detection range for FRET ...

QUEEN-based Spatiotemporal ATP Imaging in Budding and Fission Yeast

Author: Masak Takaine, date: 08/05/2019, view: 2465, Q&A: 0

[Abstract] Yeasts have provided an exceptional model for studying metabolism and bioenergetics in eukaryotic cells. Among numerous metabolites, adenosine triphosphate (ATP) is a major metabolite that is essential for all living organisms. Therefore, a clearer understanding of ATP dynamics in living yeast cells is important for deciphering cellular energy ...

Precision Technique for Splenectomy Limits Mouse Stress Responses for Accurate and Realistic Measurements for Investigating Inflammation and Immunity

Authors: Shengwen Calvin Li, Anthony D. Rangel and Mustafa H. Kabeer, date: 08/05/2019, view: 2221, Q&A: 0

[Abstract] Splenectomy in an animal model requires a standardized technique utilizing best practice to avoid variability which can result in adverse impact to the animal resulting in flawed physiologic responses simply due to technique rather than to the studied variables. In the case of the spleen, often investigators are analyzing the animal immune or ...

Unbiased Screening of Activated Receptor Tyrosine Kinases (RTKs) in Tumor Extracts Using a Mouse Phospho-RTK Array Kit

Authors: Julian Naipauer, Lucas E. Cavallin and Enrique A. Mesri, date: 04/20/2019, view: 2491, Q&A: 0

[Abstract] Kaposi’s sarcoma (KS) herpesvirus (KSHV) is a virus that causes KS, an angiogenic AIDS-associated spindle-cell neoplasm, by activating host oncogenic signaling cascades through autocrine and paracrine mechanisms. Many host signaling cascades co-opted by KSHV including PI3K/AKT/mTORC, NFkB and Notch are critical for cell-specific mechanisms of ...

Eimeria vermiformis Infection Model of Murine Small Intestine

Authors: Patrícia Figueiredo-Campos, Cristina Ferreira, Birte Blankenhaus and Marc Veldhoen, date: 12/20/2018, view: 2476, Q&A: 0

[Abstract] Eimeria vermiformis is a tissue specific, intracellular protozoan that infects the murine small intestinal epithelia, which has been widely used as a coccidian model to study mucosal immunology. This mouse infection model is valuable to investigate the mechanisms of host protection against primary and secondary infection in the small ...

H1N1 Virus Production and Infection

Authors: Binbin Zhao, Jiaoyu Shan, Rui Xiong, Ke Xu and Bin Li, date: 10/20/2018, view: 2871, Q&A: 0

[Abstract] Influenza A virus is a member of orthomyxoviridae family causing wide-spread infections in human respiratory tract. Mouse infection model is widely used in antiviral research and pathogenesis study against influenza A virus. Here, we report a protocol in infected mice with different virus doses and strains to explore how an inhibitor of ...

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: 3214, 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 ...

Assessing the Efficacy of Small Molecule Inhibitors in a Mouse Model of Persistent Norovirus Infection

Authors: Jana Van Dycke, Johan Neyts and Joana Rocha-Pereira, date: 05/05/2018, view: 3714, Q&A: 0

[Abstract] Human norovirus is the most common cause of acute gastroenteritis worldwide, resulting in estimated mortality of ~210,000 each year, of whom most are children under the age of five. However, norovirus can infect people of all age groups. There is a risk of prolonged infection in children, the elderly and patients who are immunocompromised. To ...

Detection of Protein Interactions in the Cytoplasm and Periplasm of Escherichia coli by Förster Resonance Energy Transfer

Authors: Nils Y. Meiresonne, Svetlana Alexeeva, René van der Ploeg and Tanneke den Blaauwen, date: 01/20/2018, view: 6443, Q&A: 0

[Abstract] This protocol was developed to qualitatively and quantitatively detect protein-protein interactions in Escherichia coli by Förster Resonance Energy Transfer (FRET). The described assay allows for the previously impossible in vivo screening of periplasmic protein-protein interactions. In FRET, excitation of a donor fluorescent ...

Infection of Caenorhabditis elegans with Vesicular Stomatitis Virus via Microinjection

Authors: Adam Martin, Emily A. Rex, Takao Ishidate, Rueyling Lin and Don B. Gammon, date: 11/20/2017, view: 5009, Q&A: 0

[Abstract] Over the past 15 years, the free-living nematode, Caenorhabditis elegans has become an important model system for exploring eukaryotic innate immunity to bacterial and fungal pathogens. More recently, infection models using either natural or non-natural nematode viruses have also been established in C. elegans. These models offer ...

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