title: [In vitro transgenic expression efficacy of a helper-dependent adenoviral vector encoding enhanced green fluorescent protein].
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.
authors.affiliation: Department of Immunology, Anhui Medical University, Hefei 230032, China.
authors.forename: Xianxian
authors.initials: X
authors.lastname: Zheng
authors.forename: Jinsheng
authors.initials: J
authors.lastname: He
authors.forename: Yuanhui
authors.initials: Y
authors.lastname: Fu
authors.forename: Shaohua
authors.initials: S
authors.lastname: Xu
authors.forename: Can
authors.initials: C
authors.lastname: Xie
authors.forename: Changxin
authors.initials: C
authors.lastname: Shi
authors.forename: Mei
authors.initials: M
authors.lastname: Zhang
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Wang
authors.forename: Tao
authors.initials: T
authors.lastname: Hong
mesh_terms: D000256:Adenoviridae / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics*; D006378:Helper Viruses / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D019076:Transgenes; D014760:Viral Fusion Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D014760:Viral Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins