[Construction and identification of recombinant BHV-1 expressing foot and mouth disease virus VP1 gene].
OBJECTIVE
In order to construct the recombinant bovine hepervirus-1 (BHV-1) which expressed foot and mouth disease virus (FMDV) VP1 gene, we constructed a BHV-1 gE gene transfer vector by inserting the synthetic VP1 gene of FMDV (O/China/99) under the immediate-early promoter of cytomegalovirus.


METHODS
The mixtures of parental virus (BHV-1/gE(-)/LacZ+) DNA and transfer vector was transfected into bovine turbinate cells using calcium phosphate-mediated transfection. Then the propagated viruses were harvested. The recombinant BHV-1 (designated BHV-1/gE(-)/VP1) was obtained by selection for white virus plaques.


RESULTS
PCR results showed that VP1 gene was successfully inserted into the genome of BHV-1/gE(-). The expression of VP1 in infected cells was proved by indirect immunofluorescence assay and Western blotting.


CONCLUSIONS
The research provided a basis for development of BHV-1 vector vaccines for FMD and other important bovine infectious diseases.
Ren
Xiangang
X
Division of Livestock Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China.
Xue
Fei
F
Zhu
Yuanmao
Y
Tong
Guanzhi
G
Wang
Yanhui
Y
Feng
Junke
J
Zu
Lichuang
L
Li
Jiao
J
Shi
Hongfei
H
Gao
Yuran
Y
D000818:Animals; D000914:Antibodies, Viral / Q000235:genetics; D015153:Blotting, Western; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D002417:Cattle; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003587:Cytomegalovirus / Q000235:genetics; D005536:Foot-and-Mouth Disease / Q000821:virology*; D015870:Gene Expression; D005822:Genetic Vectors; D007242:Herpesvirus 1, Bovine / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D013552:Swine; D014765:Viral Vaccines / Q000235:genetics*
D000914:Antibodies, Viral; D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins; C442501:VP1 protein, Foot-and-mouth disease virus; D014765:Viral Vaccines