[Expression of hepatitis E virus ORF3 gene fragment in baculovirus system and its immunological character].
OBJECTIVE
To express hepatitis E virus (HEV) ORF3 protein by baculovirus system and provide basis for immunological character research.


METHODS
Hepatitis E Virus ORF3 gene fragment was obtained by RT-PCR, ligated with vector pThioHisA for sequencing and then inserted into transfected vector pVL1393 to construct recombinant plasmid. Mediated by Lipofectin Reagent, the recombinant vector and baculovirus linearized DNA (BaculoGold) co-transfected insect cell Sf9 to make recombinant baculovirus. Expressed ORF3 was analyzed for its immunological character by Western blotting, and immunized Kunming Mice.


RESULTS
Recombinant ORF3 protein could be recognized by the known positive serum and promoted organism to produce HEV-specific antibody.


CONCLUSIONS
Recombinant baculovirus can express effectively HEV ORF3, which has HEV specific immunogenic character.
Du
R J
RJ
Department of Molecular Biology, Institute of Medical Biology, CAMS and PUMC, Kunming 650118, China.
Ma
Y B
YB
Tang
H
H
Zhuang
J Y
JY
Le
G Y
GY
Liu
Y
Y
Dai
C B
CB
Sun
M S
MS
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002478:Cells, Cultured; D005822:Genetic Vectors; D051379:Mice; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D014162:Transfection; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology*
C400973:ORF3 protein, Hepatitis E virus; D011994:Recombinant Proteins; D014764:Viral Proteins