Hiv Gag-igfp
Hiv Gag-igfp - Related Videos
08:21
Visualization of HIV-1 Gag Binding to Giant Unilamellar Vesicle (GUV) Membranes
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Cited by 5 •
2016
We illustrate here an in vitro membrane binding assay in which interactions between HIV-1 Gag and lipid membranes are visually analyzed using YFP-tagged Gag synthesized in a wheat germ-based in vitro translation system and GUVs prepared by an electroformation technique.
13:08
Visualizing Cell-to-cell Transfer of HIV using Fluorescent Clones of HIV and Live Confocal Microscopy
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Cited by 4 •
2010
This visualized experiment is a guide for utilizing a fluorescent molecular clone of HIV for live confocal imaging experiments.
14:23
A Restriction Enzyme Based Cloning Method to Assess the In vitro Replication Capacity of HIV-1 Subtype C Gag-MJ4 Chimeric Viruses
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Cited by 12 •
2014
HIV-1 pathogenesis is defined by both viral characteristics and host genetic factors. Here we describe a robust method that allows for reproducible measurements to assess the impact of the gag gene sequence variation on the in vitro replication capacity of the virus.
05:46
Rapid Screening of HIV Reverse Transcriptase and Integrase Inhibitors
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Cited by 15 •
2014
Here we describe cellular cytotoxicity and single round infectivity assays that allow for the rapid and accurate screening of compounds to determine their cellular cytotoxicity (CC50) and IC50 values against WT and drug resistant HIV-1.
13:58
Amplifying and Quantifying HIV-1 RNA in HIV Infected Individuals with Viral Loads Below the Limit of Detection by Standard Clinical Assays
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Cited by 10 •
2011
Quantifying levels of HIV-1 RNA in plasma and sequencing single HIV-1 genomes from individuals with viral loads below the limit of detection (50-75 copies/ml) is difficult. Here we describe how to extract and quantify plasma viral RNA using a real time PCR assay that reliably measures HIV-1 RNA down to 0.3 copies/ml and how to amplify viral genomes by single genome sequencing, from samples with very low viral loads.
FAQs
The GFP signal remains associated with Gag as the polyprotein progresses through synthesis, intracellular trafficking, plasma-membrane multimerization, and incorporation into forming virions. Fluorescence can therefore be followed across successive stages of particle formation. This continuity helps researchers connect changes in protein location with particular transitions in assembly rather than treating viral particle production as a single endpoint.
At the plasma membrane, Gag multimerization represents a key organizational step in forming viral particles. HIV Gag-iGFP makes this stage visible as a localized fluorescent signal, allowing investigators to examine where assembly concentrates and how it develops. Monitoring membrane-associated fluorescence can help separate changes in assembly organization from changes occurring earlier during Gag synthesis or trafficking.
Because fluorescence microscopy follows both signal location and signal behavior over time, HIV Gag-iGFP can show when Gag reaches relevant cellular sites and how its distribution changes during assembly. Researchers can relate intracellular trafficking to later membrane accumulation and virion incorporation. These observations provide a dynamic view of the viral structural-protein pathway instead of only a final particle measurement.
Researchers use fluorescence microscopy to monitor the reporter as Gag is synthesized, trafficked within the cell, concentrated at the plasma membrane, and incorporated into forming virions. Comparing these fluorescent patterns across the assembly sequence allows the experiment to track location and dynamics directly. The resulting observations can support quantitative analysis of assembly and budding in infection-related studies.
The reporter is useful when investigators need to determine whether an antiviral factor or candidate drug changes the viral life cycle at the level of particle assembly or budding. Fluorescent observations can reveal altered Gag distribution, membrane-associated assembly, or incorporation into forming virions. This connects treatment-related changes in cellular fluorescence with specific viral structural processes.
In immunology and infection research, the reporter provides a cellular view of how viral structural protein behavior relates to host-cell interactions. Researchers can examine assembly and budding while assessing infection-related phenotypes, creating a link between molecular events and broader infection outcomes. This makes the system useful for studying how host-associated conditions or antiviral interventions influence the viral life cycle.