In vitro transgenic expression efficacy of a helper-dependent adenoviral vector encoding enhanced green fluorescent protein / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 1108-1115, 2010.
Artículo
en Chino
| WPRIM
| ID: wpr-292164
ABSTRACT
To investigate the transgenic expressing efficacy of helper-dependent adenoviral vector (HDAd) in vitro, we constructed a HDAd encoding enhanced green fluorescent protein (EGFP), denominated as HDAd/EGFP, performed large scale preparation and purification, and then identified the purified HDAd/EGFP under fluorescent microscope and electron microscope. After the concentration of HDAd/EGFP was determined by spectrophotometer, the transgenic expression efficiency of HDAd/EGFP was compared with first generation adenoviral vector encoding EGFP (FGAd/EGFP) in vitro. Therefore, we infected A549 cells with 2000 virus particles (vp) per cell by HDAd/EGFP and FGAd/EGFP respectively and analyzed EGFP expressing level by flow cytometry. Consequently, the fluorescent expression rate and fluorescent intensity of EGFP were higher in early infected A549 cells by HDAd/EGFP than by FGAd/EGFP. HDAd, capable of expressing transgene instantly and efficiently in vitro, is a potential vaccine vector.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Adenoviridae
/
Proteínas Virales de Fusión
/
Transgenes
/
Línea Celular Tumoral
/
Proteínas Fluorescentes Verdes
/
Vectores Genéticos
/
Genética
/
Virus Helper
/
Metabolismo
Límite:
Humanos
Idioma:
Chino
Revista:
Chinese Journal of Biotechnology
Año:
2010
Tipo del documento:
Artículo
Similares
MEDLINE
...
LILACS
LIS