Selection of recombinant fowlpox virus coexpressing HIV-1 gag-gp120 and IL-6.
- Author:
Wen-zheng JIANG
1
;
Ning-yi JIN
;
Zi-jian LI
;
Li-shu ZHANG
;
Xiao-huan ZOU
;
Tie-dong WANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Antibodies, Viral; blood; Blotting, Western; Cells, Cultured; Chick Embryo; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Fibroblasts; cytology; metabolism; ultrastructure; Fowlpox; blood; immunology; virology; Fowlpox virus; genetics; immunology; Gene Products, gag; genetics; metabolism; Genetic Vectors; genetics; HIV Envelope Protein gp120; genetics; metabolism; HIV-1; genetics; metabolism; Immunization; methods; Interleukin-6; genetics; metabolism; Mice; Mice, Inbred BALB C; Microscopy, Electron; Plasmids; genetics; Recombinant Fusion Proteins; genetics; immunology; metabolism; Transfection; Viral Vaccines; genetics; immunology; metabolism
- From: Chinese Journal of Experimental and Clinical Virology 2005;19(3):267-270
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct the recombinant fowlpox virus (rFPV) coexpressing HIV-1 gag-gp120 and hIL-6.
METHODSThe recombinant expressing plasmid pUTA-GE-IL6 was successfully constructed by inserting gag-gp120 gene and hIL-6 gene into the downstream of the combined promoter ATI-p7.5 and p7.5 tandem promoter respectively. After transfecting the plasmid into chicken embryonic fibroblast (CEF) cells preinfected with FPV 282E4 strain and selecting the recombinant virus under the pressure of BUdR. The recombinant virus was analyzed by nucleic acid probe hybridization and immunoblotting. In addition, the formation of virus-like particle and the expression of interested proteins in the recombinant virus-infected p815 cells were observed, and the immunogenicity of the recombinant virus was also analyzed.
RESULTSThere was colorable dot for the positive recombinant virus, immunoblotting analysis showed that the recombinant virus could expressed both gag-gp120 and IL-6. Virus-like particles (VLP) were formed in virus-infected cells, and the interested proteins could be expressed in mammalian cells infected by the recombinant virus. The immunity index from the immunized mice showed that the recombinant virus had good immunogenicity.
CONCLUSIONThe recombinant fowlpox virus coexpressing gag-gp120 and IL-6 was successfully constructed, which may provide basis for the preparation of live vector genetic engineering vaccine and macromolecule particle vaccine against HIV-1.
