[Selection of recombinant fowlpox virus coexpressing HIV-1 gag-gp120 and IL-6].
OBJECTIVE
To construct the recombinant fowlpox virus (rFPV) coexpressing HIV-1 gag-gp120 and hIL-6.


METHODS
The 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.


RESULTS
There 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.


CONCLUSIONS
The 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.
Jiang
Wen-zheng
WZ
Key Laboratory of Genetic Engineering of PLA, Quartermaster University of PLA, Changchun 130062, China.
Jin
Ning-yi
NY
Li
Zi-jian
ZJ
Zhang
Li-shu
LS
Zou
Xiao-huan
XH
Wang
Tie-dong
TD
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D015153:Blotting, Western; D002478:Cells, Cultured; D002642:Chick Embryo; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D005586:Fowlpox / Q000097:blood / Q000276:immunology / Q000821:virology; D005587:Fowlpox virus / Q000235:genetics* / Q000276:immunology; D015683:Gene Products, gag / Q000235:genetics* / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D015699:HIV Envelope Protein gp120 / Q000235:genetics* / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism; D007114:Immunization / Q000379:methods; D015850:Interleukin-6 / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D014162:Transfection; D014765:Viral Vaccines / Q000235:genetics / Q000276:immunology / Q000378:metabolism
D000914:Antibodies, Viral; D015683:Gene Products, gag; D015699:HIV Envelope Protein gp120; D015850:Interleukin-6; D011993:Recombinant Fusion Proteins; D014765:Viral Vaccines