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.
Artigo
em Chinês
| 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:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Adenoviridae
/
Proteínas Virais de Fusão
/
Transgenes
/
Linhagem Celular Tumoral
/
Proteínas de Fluorescência Verde
/
Vetores Genéticos
/
Genética
/
Vírus Auxiliares
/
Metabolismo
Limite:
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2010
Tipo de documento:
Artigo
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