Expression of NA of influenza virus and C3d fusion gene in replication-defective recombinant adenovirus and its immune efficacy analysis.
- Author:
Feng HAN
1
;
Xuan WANG
;
Yan WANG
;
Xiao-dong ZHAO
;
Shu-hua WU
Author Information
- Publication Type:Journal Article
- MeSH: Adenoviridae; genetics; metabolism; physiology; Animals; Cloning, Molecular; Complement C3d; biosynthesis; genetics; Genetic Vectors; genetics; Immunoglobulin G; immunology; Influenzavirus A; enzymology; genetics; Mice; Mice, Inbred BALB C; Neuraminidase; biosynthesis; genetics; Recombinant Fusion Proteins; biosynthesis; genetics; Transfection; methods; Virus Replication
- From: Chinese Journal of Experimental and Clinical Virology 2013;27(1):5-7
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct a replication-defective recombinant adenovirus expressing the fusion gene of neuraminidase (NA) gene in influenza virus A/FM/1/47 and C3d and to evaluate the induced immune efficacy.
METHODSNA-C3d was cloned into shutter vector pAdTrack-CMV, which was cotransformated with adenovirus DNA into E. coli BJ5183. The recombinant adenovirus genomic DNA was generated through homological recombination. The recombinant adenovirus was produced by transfecting 293 cell line with the genomic DNA and the induced immune efficacy in mice were analyzed.
RESULTSThe integration of NA-C3d in the adenovirus genomic DNA and its expression were confirmed by PCR and Western-Blot assays respectively. After intranasal immunization, the serum IgG was induced at a titer of 1: 1000 and 1:100 000 in BALB/c mice at primary and secondary immunization respectively. The vaccinated mice were completely survived when challenged with wide influenza virus.
CONCLUSIONrecombinant adenovirus expressing NA-C3d was successfully constructed and it could induce desired immune efficacy.
