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.
Article
in Chinese
| 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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Adenoviridae
/
Viral Fusion Proteins
/
Transgenes
/
Cell Line, Tumor
/
Green Fluorescent Proteins
/
Genetic Vectors
/
Genetics
/
Helper Viruses
/
Metabolism
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2010
Type:
Article
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