Rev-nucleolar Localization Signal (NoLS)-3'Flag Mutation Expression During HIV-1 Replication
2:03
Viral Protein Lysate Collection
3:12
Rev-NoLS-3'Flag Coimmunoprecipitation
5:20
In-gel Reduction, Alkylation, and Digestion of Coomassie-stained Gel Bands
7:14
Results: Representative Nucleolar Factor Identification During HIV-1 Replication
9:05
Conclusion
Transcript
Our protocol provides alternative methods for the identification and characterization of viral, nucleolar, and non-nucleolar host factors that maintain the HIV-1 infectious cycle. The concepts used in this approach are applicable to other viral an
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Here we describe Rev immunoprecipitation in the presence of HIV-1 replication for mass spectrometry. The methods described can be used for the identification of nucleolar factors involved in the HIV-1 infectious cycle and are applicable to other disease models for the characterization of understudied pathways.