Viral Protein Promoter

The goal of this protocol is to specifically tag and selectively isolate viral DNA from infected cells for the characterization of viral genome associated proteins.

Source: Dembowski, J. A., et al. Purification of Viral DNA for the Identification of Associated Viral and Cellular Proteins. J. Vis. Exp. (2017).This video demonstrates the isolation of protein–viral DNA complexes from cell nuclei using click chemistry and magnetic bead purification. The protocol involves labeling viral genomes with alkyne-modified nucleotides, biotinylation via a click reaction, and binding to streptavidin-coated magnetic beads. Following washing and heat elution, the purified...

Begin with influenza virus-derived viral ribonucleoprotein or vRNP complexes.The vRNP complex contains an RNA polymerase bound to the viral RNA genome, essential for viral RNA synthesis.Add increasing amounts of a recombinant viral polymerase, along with a recombinant host protein, to mimic host-mediated regulation of vRNP activity.Introduce capped RNA as a primer to initiate transcription.Add nucleotide and polymerase cofactors to promote vRNP activity, and RNase inhibitors to inactivate the...

Here, we describe a newly developed multiplex fluorescent immunoassay that uses a dual-reporter flow cytometric system to concurrently detect two unique spike protein epitopes on intact severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral particles that had been captured by angiotensin-converting enzyme-2-coupled magnetic microspheres.

This video demonstrates a method for monitoring infected cell protein 0, ICP0 trafficking in herpes simplex virus-1 infection. Post-de novo synthesis, ICP0 translocates to the nucleus, later moving to the cytoplasm during infection progression. Immunofluorescence microscopy reveals and analyzes the protein's subcellular localization, offering insights into its trafficking across infection phases.