Applying Reverse Genetics to Study Measles Virus Interactions with the Host.
Methods Mol Biol
; 2808: 89-103, 2024.
Article
en En
| MEDLINE
| ID: mdl-38743364
ABSTRACT
The study of virus-host interactions is essential to achieve a comprehensive understanding of the viral replication process. The commonly used methods are yeast two-hybrid approach and transient expression of a single tagged viral protein in host cells followed by affinity purification of interacting cellular proteins and mass spectrometry analysis (AP-MS). However, by these approaches, virus-host protein-protein interactions are detected in the absence of a real infection, not always correctly compartmentalized, and for the yeast two-hybrid approach performed in a heterologous system. Thus, some of the detected protein-protein interactions may be artificial. Here we describe a new strategy based on recombinant viruses expressing tagged viral proteins to capture both direct and indirect protein partners during the infection (AP-MS in viral context). This way, virus-host protein-protein interacting co-complexes can be purified directly from infected cells for further characterization.
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Texto completo:
1
Base de datos:
MEDLINE
Asunto principal:
Proteínas Virales
/
Interacciones Huésped-Patógeno
/
Genética Inversa
/
Virus del Sarampión
Idioma:
En
Revista:
Methods Mol Biol
Asunto de la revista:
BIOLOGIA MOLECULAR
Año:
2024
Tipo del documento:
Article