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Construction and identification of helper-dependent adenoviral vector encoding hemagglutinin protein of swin-origin influenza virus a H1N1

Zhang, M.; Jiang, Y.; Chen, N.; Fu, Y.; Qiao, W.; Wang, H.; He, J.

Sheng Wu Yi Xue Gong Cheng Xue Za Zhi 31(1): 157-160

2014

ISSN/ISBN: 1001-5515
PMID: 24804504
Document Number: 673517
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.

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