Construction and characterization of a hexon-chimeric human adenovirus type 3 vector expressing one major epitope of dengue virus type 1
10.3760/cma.j.issn.0254-5101.2015.06.013
- VernacularTitle:1型登革病毒抗原表位嵌合人3型腺病毒六邻体重组病毒的构建及免疫学鉴定
- Author:
Suishan ZHAO
;
Zhichao ZHOU
;
Xiao LI
;
Ye FAN
;
Xiaohong LIAO
;
Rong ZHOU
;
Xiaobo SU
- Publication Type:Journal Article
- Keywords:
Adenovirus;
Hexon;
Hypervariable region;
Dengue virus;
Epitope
- From:
Chinese Journal of Microbiology and Immunology
2015;(6):464-468
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a recombinant human adenovirus type 3 ( HAd3 ) vector ex-pressing one major epitope of dengue virus type 1.Methods The gene encoding the envelope protein (304-314 aa) of dengue virus type 1 was inserted into the hypervariable region 1 ( HVR1 ) of HAd3 hexon by using overlap PCR.The recombinant gene was cloned into the shuttle plasmid, then linearized with AsisⅠrestriction enzyme and co-transformed into Escherichia coli BJ5183 strains with the digested backbone plas-mid for homologous recombination.The recombinant plasmid pBRAdΔE3GFP-DENV1 was transfected into AD293 cells to rescue recombinant adenovirus strains (rAdΔE3GFP-DENV1).ELISA and Western blot as-say were performed to evaluate the humoral responses induced in BALB/c mice after the immunization with rAdΔE3GFP-DENV1 strains.Results The recombinant adenovirus strains were successfully rescued. ELISA and Western blot assay showed that the antibodies in serum sample could recognize dengue virus type 1 strains.Conclusion The recombinant adenovirus strains expressing the epitope of dengue virus type 1 were successfully constructed.This study provided evidence for the development of multivalent vaccines against dengue virus.
