Combination of mass spectrometry and GST pull-down techniques to study potential interacting protein of PCV2 ORF4 / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 40-48, 2019.
Artículo
en Chino
| WPRIM
| ID: wpr-771402
ABSTRACT
A novel protein encoded by the open reading frame 4 (ORF4) was recently discovered in porcine circovirus type 2 (PCV2). However, little is known about the interaction proteins of ORF4 which hindered better understanding the biological functions of ORF4 in the life cycle of PCV2. In the present study, the ORF4 was inserted into the multiple cloning site of pCMV-N-Flag-GST, yielding recombinant plasmid pCMV-N-Flag-GST-ORF4. The recombinant plasmid was transfected into 293T cells and the intracellular interaction complex of ORF4 were enriched and separated by GST pull-down and SDS-PAGE, sequentially. The potential interacting proteins of PCV2 ORF4 were stained with silver and identified by mass spectrometry (MS). Finally, five candidate ORF4-interacting proteins, including Serine/threonine-protein phosphatase 6 catalytic subunit, alpha cardiac muscle 1, actin, SEC14-like protein 5 and myosin 9 were identified. These results would benefit a better understanding of the biological function of ORF4 in PCV2 infected cells.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Espectrometría de Masas
/
Porcinos
/
Proteínas Virales
/
Sistemas de Lectura Abierta
/
Circovirus
/
Infecciones por Circoviridae
/
Células HEK293
Límite:
Animales
/
Humanos
Idioma:
Chino
Revista:
Chinese Journal of Biotechnology
Año:
2019
Tipo del documento:
Artículo
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