Construction, expression and location of eukaryotic expression vector CMV-FLAG-RB in human prostate cancer PC-3 cells / 南方医科大学学报
Journal of Southern Medical University
; (12): 422-425, 2010.
Article
en Zh
| WPRIM
| ID: wpr-269538
Biblioteca responsable:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To construct an eukaryotic recombinant expression vector for retinoblastoma 1 gene (RB-1) and investigate the role of RB-1 in prostate cancer.</p><p><b>METHODS</b>The coding sequence of RB-1 gene tagged with FLAG was amplified from the plasmid CMV-RB by PCR method. The fragment was cloned into CMV expression vector and identified by restriction enzyme digestion and sequence analysis. Western Blotting was used to detect RB-1 expression and immunofluorescence was used to observe RB-1 distribution in PC-3 cells transfected with the recombinant.</p><p><b>RESULTS</b>The expression vector CMV-FLAG-RB was successfully constructed as confirmed by PCR, endonuclease digestion and DNA sequence analysis. RB-1 protein was highly expressed and showed a nuclear distribution in PC-3 cells transfected with the recombinant.</p><p><b>CONCLUSIONS</b>The eukaryotic expression vector for RB-1 has been successfully constructed and can be efficiently expressed in PC-3 cells. The expression of RB-1 is located in the cell nuclei.</p>
Texto completo:
1
Base de datos:
WPRIM
Asunto principal:
Patología
/
Neoplasias de la Próstata
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Proteínas Recombinantes
/
Datos de Secuencia Molecular
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Secuencia de Bases
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Transfección
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Expresión Génica
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Proteína de Retinoblastoma
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Clonación Molecular
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Línea Celular Tumoral
Límite:
Humans
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Male
Idioma:
Zh
Revista:
Journal of Southern Medical University
Año:
2010
Tipo del documento:
Article