Construction and identification of helper-dependent adenoviral vector encoding hemagglutinin protein of swin-origin influenza virus A H1N1.
- Author:
Mei ZHANG
;
Yanze JIANG
;
Nianhua CHEN
;
Yuanhui FU
;
Wei QIAO
;
He WANG
;
Jinsheng HE
- Publication Type:Journal Article
- MeSH:
Adenoviridae;
Cell Line;
Cloning, Molecular;
Genetic Vectors;
Helper Viruses;
Hemagglutinin Glycoproteins, Influenza Virus;
biosynthesis;
Humans;
Influenza A Virus, H1N1 Subtype
- From:
Journal of Biomedical Engineering
2014;31(1):157-160
- CountryChina
- Language:Chinese
-
Abstract:
Abstract: In order to investigate immune protection against swine-origin influenza virus (S-OIV) A H1N1, the helper-dependent adenovirus vector (HDAd) system was exploited to construct recombinant HDAd encoding hemagglutinin (HA). The HA gene was synthesized and cloned to the HDAd backbone. Then, the HDAd/HA DNA molecules were transfected into 293Cre4 cells with calcium phosphate. The cells were infected by helper virus 16 hours after the transfection. The 293Cre4 cells were coinfected with HDAd/HA and the helper virus for large-scale preparation of HDAd/HA. The HDAd/HA was obtained and purified twice with CsCI density ultracentrifugation and observed morphologically under transmission electron microscope, and the expression of HA protein was analyzed with RTPCR. Recombinant HDAd/HA expressing HA protein was successfully constructed which could pave the way for in vivo investigation on immunogenicity and efficacy against S-OIV A H1N1 infection.
