In vitro transgenic expression efficacy of a helper-dependent adenoviral vector encoding enhanced green fluorescent protein.
- Author:
Xianxian ZHENG
1
;
Jinsheng HE
;
Yuanhui FU
;
Shaohua XU
;
Can XIE
;
Changxin SHI
;
Mei ZHANG
;
Xiaobo WANG
;
Tao HONG
Author Information
1. Department of Immunology, Anhui Medical University, Hefei 230032, China.
- Publication Type:Journal Article
- MeSH:
Adenoviridae;
genetics;
metabolism;
Cell Line, Tumor;
Genetic Vectors;
genetics;
Green Fluorescent Proteins;
genetics;
Helper Viruses;
genetics;
metabolism;
Humans;
Transgenes;
Viral Fusion Proteins;
genetics;
metabolism
- From:
Chinese Journal of Biotechnology
2010;26(8):1108-1115
- CountryChina
- Language:Chinese
-
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.
